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  • 1990-1994  (275)
  • 1955-1959  (4)
  • 1930-1934
  • 1890-1899
  • gene expression
  • 101
    Electronic Resource
    Electronic Resource
    Upstream regulatory sequences from two β-conglycinin genes (1993)
    Springer
    Plant molecular biology 22 (1993), S. 873-885 
    Details
    ISSN: 1573-5028
    Keywords: β-conglycinin ; gene expression ; legumin box ; seed storage protein ; vicilin box
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Genes encoding the β-conglycinin seed storage proteins of soybean are expressed only in seeds during specific stages of development. The different subunits of β-conglycinin, α′, α and β, are encoded by distinct members of a gene family. Yet there are marked differences in the regulation of the genes encoding the α′/α and β subunits. Previous work (Chen et al., EMBO J 7: 297–302, 1988) identified a seed specific transcriptional enhancer upstream of a gene encoding the α′ subunit. Mutations were made within this region to discern its functional components. Among those identified is a 62 bp region (between −77 and −140) that contains a vicilin box consensus sequence as well as a sequence that binds the soybean nuclear factor SEF4 in vitro. A second region, which contains a sequence homologous to the core of the legumin box consensus (i.e., CATGCAT-like or RY repeat element) at −246, was also shown to affect the activity of this enhancer in transgenic plants. A series of 5′ terminal deletions were used to identify regulatory elements upstream of the β subunit gene. Two regions were identified (from −553 to −442 and from −308 to −72) that, when deleted, led to a marked reduction in gene expression. Both of these elements contain sequences that bind SEF4 in vitro. The distal element also contains an AT-rich segment that recognizes a second nuclear factor, SEF1, in vitro. Neither of these elements contains any homology to the vicilin box consensus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027372
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  • 102
    Electronic Resource
    Electronic Resource
    Molecular characterization of four chitinase cDNAs obtained fromCladosporium fulvum-infected tomato (1993)
    Springer
    Plant molecular biology 22 (1993), S. 1017-1029 
    Details
    ISSN: 1573-5028
    Keywords: chitinase ; gene expression ; pathogenesis-related protein ; plant-fungus interaction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Complementary DNA clones encoding acidic and basic isoforms of tomato chitinases were isolated fromCladosporium fulvum-infected leaves. The clones were sequenced and found to encode the 30 kDa basic intracellular and the 26 and 27 kDa acidic extracellular tomato chitinases previously purified (M.H.A.J. Joostenet al., in preparation). A fourth truncated cDNA which appears to encode an extracellular chitinase with 82% amino acid similarity to the 30 kDa intracellular chitinase was also isolated. Characterization of the clones revealed that the 30 kDa basic intracellular protein is a class I chitinase and that the 26 and 27 kDa acidic extracellular proteins which have 85% peptide sequence similarity are class II chitinases. The characterized cDNA clones represent four from a family of at least six tomato chitinases. Southern blot analysis indicated that, with the exception of the 30 kDa basic intracellular chitinase, the tomato chitinases are encoded by one or two genes. Northern blot analysis showed that the mRNA encoding the 26 kDa acidic extracellular chitinase is induced more rapidly during an incompatibleC. fulvum-tomato interaction than during a compatible interaction. This difference in timing of mRNA induction was not observed for the 30 kDa basic intracellular chitinase.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028974
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  • 103
    Electronic Resource
    Electronic Resource
    Down-regulation of two non-homologous endogenous tomato genes with a single chimaeric sense gene construct (1993)
    Springer
    Plant molecular biology 23 (1993), S. 1-9 
    Details
    ISSN: 1573-5028
    Keywords: down-regulation ; gene expression ; polygalacturonase ; pectinesterase ; chimaeric sense gene ; transgenic ; co-suppression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tomatoes (Lycopersicon esculentum Mill cv. Ailsa Craig) were transformed with a gene construct having 244 bp of the 5′ end of a polygalacturonase (PG) cDNA, coding for a 71 amino acid N-terminal extension to the mature protein, fused to 1320 bp of a pectinesterase (PE) cDNA encoding the full sequence of the mature PE protein. This chimaeric gene was inserted in a sense orientation between a CaMV 35S promoter and terminator for constitutive expression. In transformed tomato plants expression of the endogenous PG and PE genes in the fruit was inhibited; there was little or no observable PG and PE mRNA and a substantial reduction in the level of PG and PE enzyme activity. The transgene was expressed in the leaves of the transformed plants as demonstrated by the accumulation of mRNA, but no protein product could be identified. However, no transgene mRNA or protein were observed in the transgenic fruit. This paper represents the first report of the down-regulation of two non-homologous endogenous genes using a single gene construct. A sense gene construct was responsible for these effects. These findings are discussed in relation to possible mechanisms of action of co-suppression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021414
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  • 104
    Electronic Resource
    Electronic Resource
    A histidine decarboxylase-like mRNA is involved in tomato fruit ripening (1993)
    Springer
    Plant molecular biology 23 (1993), S. 627-631 
    Details
    ISSN: 1573-5028
    Keywords: ethylene ; histamine ; gene expression ; Lycopersicon esculentum ; ripening mutant ; ripening inhibitor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract DNA sequencing of a tomato ripening-related cDNA, TOM 92, revealed an open reading frame with homology to several pyridoxal 5′-phosphate histidine decarboxylases, containing the conserved amino acid residues known to bind pyridoxal phosphate and α-fluoromethylhistidine, an inhibitor of enzyme activity. TOM 92 mRNA accumulated during early fruit ripening and then declined. Fruit of the ripeningimpaired tomato mutant, ripening inhibitor (rin), did not accumulate TOM 92 mRNA, and its accumulation was not restored by treatment of fruit with ethylene. The TOM 92 mRNA was not detected in tomato leaves and unripe fruit.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019310
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  • 105
    Electronic Resource
    Electronic Resource
    Solanum phureja genes are expressed in the leaves and tubers of aneusomatic potato dihaploids (1993)
    Springer
    Euphytica 69 (1993), S. 1-6 
    Details
    ISSN: 1573-5060
    Keywords: gene expression ; isozymes ; patatin ; potato dihaploids ; Solanum phureja
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The expression of leaf isozymes and tuber patatin in dihaploids derived from the Solanum tuberosum cv. Pentland Crown was investigated. Seven of the dihaploids were aneusomatic containing additional chromosomes from the S. phureja dihaploid inducer. Of these, four genotypes expressed leaf isozymes characteristic of the S. phureja dihaploid inducer, and the tubers of three aneusomatic dihaploids contained a S. phureja form of patatin. Aneusomatic dihaploids in which the proportion of cells containing additional S. phureja chromosomes was relatively small (i.e. 1–15%) did not express leaf isozyme markers or patatin bands characteristic of the dihaploid inducer or showed only faint expression of one or two markers. However, those with a high proportion of cells containing additional chromosomes (50–55%) had a range of strongly expressed leaf isozymes that were characteristic of the dihaploid inducer and also expressed the S. phureja tuber patatin. One dihaploid genotype was exclusively euploid (2n〈24), yet is expressed a S. phureja leaf isozyme marker and S. phureja tuber patatin, suggesting recombination or chromosome substitution between the genome of the S. phureja dihaploid inducer and the cultivar Pentland Crown.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021720
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  • 106
    Electronic Resource
    Electronic Resource
    Developmental expression of glutathione-S-transferase in maize and its possible connection with herbicide tolerance (1993)
    Springer
    Euphytica 67 (1993), S. 221-230 
    Details
    ISSN: 1573-5060
    Keywords: Zea mays ; glutathione-S-transferase ; glutathione ; herbicide tolerance ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Crop improvement for tolerance to specific herbicides is an important breeding target, since molecules performing well with regard to environmental safety are frequently not completely selective for crops. The glutathione (GSH)/glutathione-S-transferase (GST) system is a general mechanism of detoxification that in higher plants may confer tolerance to some herbicides. GSH level and GST activity were measured in different maize inbred lines, in the absence or in the presence of EPTC (a thiocarbamate) and of Alachlor (a chloroacetanilide); a wide genetic variability was observed for these parameters, which appear to be involved in plant tolerance to herbicides. Isozyme analysis was performed on roots, leaves, scutellum, pollen, coleoptile, mesocotyl of the same inbreds: it revealed the presence of many GST forms in maize, showing high polymorphism; they are controlled by at least five genes, the expression of which is developmentally regulated in the different tissues analyzed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040624
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  • 107
    Electronic Resource
    Electronic Resource
    Redox control of gene expression and the function of chloroplast genomes — an hypothesis (1993)
    Springer
    Photosynthesis research 36 (1993), S. 95-102 
    Details
    ISSN: 1573-5079
    Keywords: chloroplast genome ; electron transport ; evolution ; gene expression ; redox response regulators ; redox sensors
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Two-component regulatory systems that respond to changes in redox potential have recently been discovered in bacteria. ‘Redox sensors’ are defined as electron carriers which initiate control of gene expression upon oxidation or reduction. ‘Redox response regulators’ are defined as DNA-binding proteins which modify gene expression as a result of the action of redox sensors. Redox sensors and redox response regulators may comprise a mechanism for feedback control of redox potential in photosynthetic electron transport chains, thereby protecting plants, algae and photosynthetic bacteria from damage caused by electrochemistry operating on inappropriate electron donors and acceptors. Chloroplast redox sensors and redox response regulators, themselves encoded in the nucleus, may place chloroplast gene expression under redox regulatory control. This may account for the persistence, in evolution, of chloroplast genomes, and for the constancy of the sub-set of chloroplast proteins encoded and synthesised in situ. These and other predictions are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00016274
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  • 108
    Electronic Resource
    Electronic Resource
    Jasmonic acid-inducible gene expression of a Kunitz-type proteinase inhibitor in potato tuber disks (1993)
    Springer
    Plant molecular biology 21 (1993), S. 539-541 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; jasmonic acid ; Kunitz-type proteinase inhibitor ; patatin ; potato (Solanum tuberosum L.) ; wounding
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Messenger RNAs of a potato (Solanum tuberosum L.) Kunitz-type proteinase inhibitor(s) (PKPI) were present in potato disks excised from tubers stored for 14 months (old tubers) or 2 months (young tubers) after harvest, and disappeared during the aseptic culture. The PKPI mRNA accumulation was found to be induced in potato disks from the old tubers by the addition of jasmonic acid (JA) [3-oxo-2-(2′-cis-pentenyl)-cyclopentane-1-acetic acid].
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028810
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  • 109
    Electronic Resource
    Electronic Resource
    Molecular cloning and characterization of a legumin-like storage protein cDNA of Douglas fir seeds (1993)
    Springer
    Plant molecular biology 21 (1993), S. 709-715 
    Details
    ISSN: 1573-5028
    Keywords: cDNAs ; seed storage proteins ; Pseudotsuga menziesii ; gene expression ; nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA library was made from poly(A)+ RNA isolated from developing Douglas fir (Pseudotsuga menziesii) embryo and megagametophytic tissue, and the cDNA clones were identified by immunoscreening with polyclonal antiserum against the crystalloid storage protein complex of Douglas fir. The nucleotide sequence of the longest cDNA insert (DF1) was analysed. The amino acid sequence derived from the DNA sequence verified its identity as a legumin-like storage protein (pseudotsugin) and confirmed that the protein is synthesized as a precursor similar to the 11–12S storage globulins. The transcripts corresponding to cDNA insert DF1 were abundant in the early-to mid-stages of embryogenesis in the diploid embryonic axes as well as in the haploid megagametophytic tissue. The deduced amino acid sequence of pseudotsugin consists of a 29 amino acid N-terminal signal peptide preceding the acidic polypeptide region (286 amino acids) and the subsequent basic polypeptide region (212 amino acids). The site for post-transcriptional cleavage of the precursor polypeptide to make the A and B polypeptides is localized between asparagine −315 and glycine −316 and is highly conserved between angiosperms and gymnosperms. The deduced amino acid sequence for the DF1 cDNA clone reveals that pseudotsugin is rich in arginine, glutamic acid and serine and is low in cysteine, methionine and lysine. Comparison of the deduced amino acid sequence of Douglas fir pseudotsugin shows between 29–38.5% identity with angiosperm species, 63% identity with interior spruce, and 60% identity with eastern white pine.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014555
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  • 110
    Electronic Resource
    Electronic Resource
    Expression of an ABA-responsive osmotin-like gene during the induction of freezing tolerance in Solanum commersonii (1993)
    Springer
    Plant molecular biology 21 (1993), S. 729-735 
    Details
    ISSN: 1573-5028
    Keywords: ABA ; cDNA cloning ; freezing tolerance ; gene expression ; osmotin-like protein ; Solanum commersonii
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated a cDNA (pA13) of an ABA-responsive gene from suspension cultures of Solanum commersonii. The deduced amino acid sequence of pA13 cDNA revealed 89 and 91% identity with tobacco osmotin and tomato NP24 protein, respectively. The accumulation of the transcript corresponding to pA13 cDNA was regulated by ABA, cold temperature, and low water potential treatments. Cold-induced accumulation of the pA13 transcript was partially suppressed by fluridone, an ABA synthesis inhibitor, and the suppression was restored by exogenous ABA application. The transcript corresponding to pA13 also accumulated in an organ-specific manner in response to ABA or cold treatment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014558
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  • 111
    Electronic Resource
    Electronic Resource
    Cis-acting regulatory regions of the soybean seed storage 11S globulin gene and their interactions with seed embryo factors (1993)
    Springer
    Plant molecular biology 21 (1993), S. 973-984 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; Glycine max ; protein-DNA interaction ; seed storage protein gene ; transcriptional regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A 2.2 kb fragment containing the 5′-flanking region of the soybean glycinin A2B1a gene and its successive deletions with a shorter 5′-flanking sequence were fused, in frame, to the β-glucuronidase (GUS) reporter gene. The resultant fusions were introduced into tobacco plants via Agrobacterium tumefaciens. Assays of the GUS activity in seeds of transgenic tobacco showed that the upstream region, −657 to −327 (relative to the transcription initiation site [+1]), of the glycinin gene is required for optimal expression of the transformed gene. Interactions between embryo nuclear factors and DNA fragments covering the downstream region of −326, in which are included the TATA box and legumin boxes, were not apparent. The embryo factors capable of binding specifically to three subregions, −653 to −527, −526 to −422, and −427 to −321, of the upstream regulatory region were detected. Such factors appeared to be organ-specific and could be found solely in developing seeds at the early middle stage of embryogenesis (around 24 days after flowering). Evidence obtained by characterizing the nature of the binding proteins and by gel mobility shift assays established that the same factor does interact with a consensus motif 5′-ATA/TATTTCN-/CTA-3′ which occurs four times in the cis-acting regulatory region between −657 and −327. Moreover, this conserved motif could also be found in the 5′ regulatory region of another glycinin A1aB1b gene. Thus it is likely that the observed interaction between the nuclear factor and the conserved motifs would lead to activation of transcription from the glycinin genes in maturing soybean seeds.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023596
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  • 112
    Electronic Resource
    Electronic Resource
    Identification of cis-acting elements involved in the regulation of the pathogenesis-related gene STH-2 in potato (1993)
    Springer
    Plant molecular biology 22 (1993), S. 279-291 
    Details
    ISSN: 1573-5028
    Keywords: wounding ; elicitor ; plant defense ; gene expression ; tuber ; Solanum tuberosum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have characterized a genomic clone containing the potato pathogenesis-related genes STH-2 and STH-21. The two genes are found 4 kb apart on the same chromosome and their sequences are highly similar. They present the same transcriptional orientation and are both interrupted by a single intron. A chimaeric gene consisting of 1015 bp of 5′-flanking sequence and part of the first exon of STH-2 fused to the bacterial β-glucuronidase gene was highly-expressed in tubers of transgenic potato plants after wounding and elicitor treatments. The levels of activity observed in these transgenic plants parallel those observed for the accumulation of STH-2 mRNAs under similar conditions. This indicates that cis-acting elements necessary for the proper activation of the gene are present within 1 kb of 5′-flanking sequences. Functional analysis of 5′ deletions of the STH-2/GUS constructs by transient expression in leaf protoplasts revealed the presence of an upstream regulatory sequence between -135 and -52 which contains a TGAC motif, and a possible negative regulatory region between -52 and -28. A factor present in nuclear extracts of wounded potato tubers was found to bind specifically to nucleotides located between -135 to -105, suggesting that this region contains important cis-regulatory elements.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014935
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  • 113
    Electronic Resource
    Electronic Resource
    Structure and expression of a nuclear gene for the PSI-D subunit of photosystem I inNicotiana sylvestris (1993)
    Springer
    Plant molecular biology 22 (1993), S. 985-994 
    Details
    ISSN: 1573-5028
    Keywords: ferredoxin-binding subunit ; gene expression ; gene structure ; Nicotiana sylvestris ; photosystem I ; psaD
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The PSI-D subunit is the ferredoxin-binding site of photosystem I, and is encoded by the nuclear genepsaD. We isolated apsaD genomic clone fromNicotiana sylvestris, by screening a genomic library with apsaD cDNA which we previously cloned fromN. sylvestris (Yamamotoet al., Plant Mol Biol 17: 1251, 1991). Nucleotide sequence analysis revealed that this genomic clone contains apsaD gene, which does not correspond to thepsaD cDNA, so we designated these genespsaDb andpsaDa, respectively. ThepsaDb clone encodes a protein of 214 amino acids uninterrupted by introns. The N-terminal sequence determined for theN. sylvestris PSI-D protein encoded bypsaDb begins at the 49th residue. The products ofpsaDa andpsaDb share 82.7% and 79.5% identity at the amino acid and nucleotide levels, respectively. Genomic Southern analysis showed that two copies ofpsaD are present in theN. sylvestris genome. Ribonuclease protection assays and immunoblot analysis inN. sylvestris indicate that both genes are expressed in leaves, stems and flower buds, but neither is expressed in roots. During leaf development, the ratio ofpsaDb topsaDa mRNA increases from 0.12 in leaf buds to 0.36 in mature leaves. The relative abundance of the corresponding proteins decreased over the same developmental period. These results indicate that differential regulation mechanisms controlpsaDa andpsaDb expression at both the mRNA and protein levels during leaf development.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028971
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  • 114
    Electronic Resource
    Electronic Resource
    The resistance of cowpea seeds to bruchid beetles is not related to levels of cysteine proteinase inhibitors (1993)
    Springer
    Plant molecular biology 23 (1993), S. 215-219 
    Details
    ISSN: 1573-5028
    Keywords: cowpea ; cystatin ; cysteine endoproteinases ; gene expression ; insect resistance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA encoding a cysteine proteinase inhibitor was isolated from a cDNA library prepared from developing seeds of an insect-resistant line of cowpea. The sequence of the encoded protein was homologous with those of other plant cysteine endoproteinase inhibitors, and with Type 2 cystatins from animals. Southern blot analyses indicated that small gene families were present in both resistant and susceptible lines of cowpea, while northern blot analyses showed similar levels of expression. It is concluded that the levels of expression of the inhibitor do not account for the differences in insect resistance of the two lines.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021433
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  • 115
    Electronic Resource
    Electronic Resource
    cDNA cloning and characterisation of novel ripening-related mRNAs with altered patterns of accumulation in the ripening inhibitor (rin) tomato ripening mutant (1993)
    Springer
    Plant molecular biology 23 (1993), S. 193-207 
    Details
    ISSN: 1573-5028
    Keywords: dehydrogenase ; ethylene ; fruit ; gene expression ; UDP glucuronosyl/glucosyl transferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA library produced from mRNA isolated from the pericarp of wild-type tomato fruit (Lycopersicon esculentum Mill. cv Ailsa Craig) at the first visible sign of fruit ripening was differentially screened to identify clones whose homologous mRNAs were present at reduced levels in fruit of the tomato ripening mutant, ripening inhibitor,rin. Five clones were isolated (pERT 1, 10, 13, 14, 15). Accumulation of mRNA homologous to each of these clones increased during the ripening of wild-type fruit and showed reduced accumulation in ripening rin fruit. The levels of three of them (homologous to ERT 1, 13 and 14) were increased by ethylene treatment of the mutant fruit. A further clone, ERT 16 was identified for a mRNA present at a high level in both normal and mutant fruit at early stages of ripening. Database searches revealed no significant homology to the DNA sequence of ERT 14 and 15; however, DNA and derived amino acid sequence of ERT 1 both contain regions of homology with several reported UDP-glucosyl and glucuronosyl transferases (UDPGT) and with a conserved UDPGT motif. A derived amino acid sequence from the ERT 10 cDNA contains a perfect match to a consensus sequence present in a number of dehydrogenases. The ERT 13 DNA sequence has homology with an mRNA present during potato tuberisation. The presence of these mRNAs in tomato fruit is unreported and their role in ripening is unknown. The ERT 16 DNA sequence has homology with a ripening/stress-related cDNA isolated from tomato fruit pericarp.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021431
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  • 116
    Electronic Resource
    Electronic Resource
    Molecular characterization of maize extensin expression (1993)
    Springer
    Plant molecular biology 23 (1993), S. 685-695 
    Details
    ISSN: 1573-5028
    Keywords: extensin ; gene expression ; hydroxyproline ; maize ; silk ; vegetative tissues
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract This study concerned the developmental regulation of wall-localized, hydroxyproline-containing proteins in maize tissues and organs. Silk and pericarp cell walls contained more peptidyl hydroxyproline than did walls of any vegetative tissue, although all tissues and organs accumulated these proteins as they matured. In many tissues, hydroxyproline-rich proteins are first associated with the wall in a soluble form before being insolubilized through covalent attachment to the matrix. Because hydroxyproline was more soluble earlier than later in development, it appears that insolubilization was occurring in maize tissues and organs as well. Tissue prints reacted with an anti-extensin antibody gave positive results, indicating the presence of a soluble form of this common hydroxyproline-rich glycoprotein (HRGP). Silk and pericarp cells actively synthesized this extensin from abundant transcripts. In vegetative tissues, extensin transcripts were somewhat more abundant in seedlings than in pre-anthesis or mature plants, but levels were much lower than in silk and pericarp. Southern blots of maize genomic DNA indicated that these extensin transcripts are encoded by a small multigene family. Potential roles for extensin in reproductive/protective tissues versus the embryo or vegetative tissues are suggested.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021524
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  • 117
    Electronic Resource
    Electronic Resource
    Metabolic regulation of α-amylase gene expression in transgenic cell cultures of rice (Oryza sativa L.) (1993)
    Springer
    Plant molecular biology 23 (1993), S. 737-747 
    Details
    ISSN: 1573-5028
    Keywords: α-amylase ; transgenic cell culture ; gene expression ; metabolic regulation ; Oryza sativa ; transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Expression of two genes in the α-amylase gene family is controlled by metabolic regulation in rice cultured cells. The levels of RAmy3D and RAmy3E mRNAs in rice cultured cells are inversely related to the concentration of sugar in the culture medium. Other genes in the rice α-amylase gene family have little or no expression in cultured cells; these expression levels are not controlled by metabolic regulation. A RAmy3D promoter/GUS gene fusion was metabolically regulated in the transgenic rice cell line 3DG, just as the endogenous RAmy3D gene is regulated. An assay of GUS enzyme activity in 3DG cells demonstrated that RAmy3D/GUS expression is repressed when sugar is present in the culture medium and induced when sugar is removed from the medium. The 942 bp fragment of the RAmy3D promoter that was linked to the coding region of the GUS reporter gene thus contains all of the regulatory sequences necessary for metabolic regulation of the gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021529
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  • 118
    Electronic Resource
    Electronic Resource
    Treatment of Agrobacterium or leaf disks with 5-azacytidine increases transgene expression in tobacco (1993)
    Springer
    Plant molecular biology 21 (1993), S. 429-435 
    Details
    ISSN: 1573-5028
    Keywords: azacytidine ; DNA methylation ; gene expression ; inactivation ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have studied the effect of the demethylating agent azacytidine (azaC) on expression of a β-glucuronidase (GUS) gene transferred to tobacco leaf disks by Agrobacterium-mediated transformation. In a system where no selection was performed, where shoot formation was partially repressed, and where Agrobacterium does not express the GUS gene, we were able to follow the early events of transient and stable expression. Two days after inoculation, 8% of the cells expressed GUS but this proportion rapidly decreased to near zero in the following week. Treatment of leaf disks with azaC just after transformation retarded this inactivation to some extent, while treatment of Agrobacterium prior to transformation increased the frequency of transient expression. Three weeks after inoculation the number of GUS-expressing cells increased 4- to 6-fold in the leaf disks treated with azaC and in the leaf disks transformed with azaC-treated bacteria, while the control remained low. These data suggest that DNA methylation is involved in transgene inactivation and that a large number of silent but potentially active transgenes become integrated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028801
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  • 119
    Electronic Resource
    Electronic Resource
    Transient and stable expression ofgusA fusions with rice genes in rice, barley and perennial ryegrass (1993)
    Springer
    Plant molecular biology 22 (1993), S. 1101-1127 
    Details
    ISSN: 1573-5028
    Keywords: transformation ; promoters ; introns ; gene expression ; Oryza sativa ; Hordeum vulgare ; Lolium perenne
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcriptional and translational fusions were made between the reading frame coding for β-D-glucuronidase and sequences of either a constitutively expressed rice gene (GOS2) involved in initiation of translation or a light-inducible rice gene (GOS5). The transient expression of the fusions was studied via particle bombardment of seedling tissues of rice, perennial ryegrass and barley. Furthermore, the results of transient and stable expression were compared for cell suspensions of four rice varieties, one barley variety and one perennial ryegrass variety. TheGOS2-gusA fusions were active in all three monocots studied. Best results were obtained for a construct having both a transcriptional and a translational fusion as well as intron and exon sequences (PORCEHyg). The level of GUS activity was in the range of activities as obtained by the 35S CaMV promoter transcriptionally fused togusA. ThegusA fusion with the light-inducible gene (GOS5) was active in green seedling tissues of all monocots studied. Also a weak expression compared to theGOS2 constructs was found in stably transformed rice callus. ThegusA fusions with the mannopine synthase promoters 1′ and 2′ of the TR-DNA were transiently expressed at lower levels in cell suspensions than PORCEHyg. For stably transformed rice callus the expression of theGOS2-gusA fusion often decreased during prolonged subculture. This decrease in GUS activity and the various GUS-staining phenotypes of transgenic calli are explained by the presence of different cell types in the suspensions used and in the calli. It is presumed that the nature of the cells and their relative contribution in the calli change drastically upon further subculture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028980
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    Circadian and light-regulated expression of nitrate reductase in Arabidopsis (1993)
    Springer
    Plant molecular biology 23 (1993), S. 349-364 
    Details
    ISSN: 1573-5028
    Keywords: circadian regulation ; enzyme activity ; gene expression ; light regulation ; nitrate reductase ; phytochrome
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of a number of plant genes is regulated by an endogenous circadian clock. We report that the Arabidopsis NIA2 (nitrate reductase) gene shows robust circadian oscillations in mRNA accumulation which persist for at least 5 days in plants that have been grown in a light-dark (LD) cycle and then transferred to continuous light (LL). We further show that NIA2 mRNA accumulation oscillates in a circadian fashion in plants that have been grown in LD and then transferred to continuous darkness (DD). Results from nuclear run-on transcriptional analysis suggest that the oscillations in steady-state levels of NIA2 mRNA abundance are not primarily due to changes in transcription but, instead, reflect post-transcriptional regulation. The circadian oscillations in NIA2 mRNA abundance are paralleled by circadian oscillations in nitrate reductase enzyme activity (NR activity) in Arabidopsis plants that have been grown in LD and then transferred either to DD or to LL. Etiolated Arabidopsis seedlings express neither NIA2 mRNA nor NR activity. However, both NIA2 mRNA accumulation and NR activity are induced by exposure to white light. The inductive effects of light on NIA2 mRNA accumulation are due, at least in part, to a very low fluence phytochrome-mediated response. However, the persistence of circadian oscillations in NIA2 mRNA abundance for at least 5 days in LL demonstrates that the circadian clock is capable of overriding or gating the inductive effects of light on NIA2 mRNA accumulation in Arabidopsis for an extended, continuous period of time.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029010
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    Coordinate gene expression of five subclass histones and the putative transcription factors, HBP-1a and HBP-1b, of histone genes in wheat (1993)
    Springer
    Plant molecular biology 23 (1993), S. 429-434 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; germination ; transcription factor ; wheat histone gene ; wheat seedling
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of genes encoding five histones (H1, H2A, H2B, H3 and H4) and the putative transcription factors HBP-1a (17) and HBP-1b (c38) was examined during early germination and in various tissues of young wheat seedlings. The steady-state levels of core histone (H2A, H2B, H3 and H4) mRNAs were coordinately cell cycle-dependent and paralleled the rate of DNA synthesis during early germination, whereas the expression pattern of the linker histone (H1) genes differed. The five subclass histone genes were actively expressed in the meristematic tissues of young seedlings. Moreover, H1 genes were expressed in leaves that consist mostly of non-proliferating cells, in which core histone genes showed little expression. Quantitative alterations to the mRNAs of the putative transcription factors HBP-1a (17) and HBP-1b (c38) of wheat histone genes were similar to those of the core histone mRNAs, suggesting that both factors function in the cell cycle-dependent expression of wheat core histone genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029019
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  • 122
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    Transient and stable expression of gusA fusions with rice genes in rice, barley and perennial ryegrass (1993)
    Springer
    Plant molecular biology 23 (1993), S. 643-669 
    Details
    ISSN: 1573-5028
    Keywords: transformation ; promoters ; introns ; gene expression ; Oryza sativa ; Hordeum vulgare ; Lolium perenne
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcriptional and translational fusions were made between the reading frame coding for β-D-glucuronidase and sequences of either a constitutively expressed rice gene (GOS2) involved in initiation of translation or a light-inducible rice gene (GOS5). The transient expression of the fusions was studied via particle bombardment of seedling tissues of rice, perennial ryegrass and barley. Furthermore, the results of transient and stable expression were compared for cell suspensions of four rice varieties, one barley variety and one perennial ryegrass variety. The GOS2-gusA fusions were active in all three monocots studied. Best results were obtained for a construct having both a transcriptional and a translational fusion as well as intron and exon sequences (PORCEHyg). The level of GUS activity was in the range of activities as obtained by the 35S CaMV promoter transcriptionally fused to gusA. The gusA fusion with the light-inducible gene (GOS5) was active in green seedling tissues of all monocots studied. Also a weak expression compared to the GOS2 constructs was found in stably transformed rice callus. The gusA fusions with the mannopine synthase promoters 1′ and 2′ of the TR-DNA were transiently expressed at lower levels in cell suspensions than PORCEHyg. For stably transformed rice callus the expression of the GOS2-gusA fusion often decreased during prolonged subculture. This decrease in GUS activity and the various GUS-staining phenotypes of transgenic calli are explained by the presence of different cell types in the suspensions used and in the calli. It is presumed that the nature of the cells and their relative contribution in the calli change drastically upon further subculture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021522
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    Molecular analysis and spatial expression pattern of a low-temperature-specific barley gene, blt101 (1993)
    Springer
    Plant molecular biology 23 (1993), S. 871-879 
    Details
    ISSN: 1573-5028
    Keywords: cold ; low temperature ; barley ; gene expression ; cDNA ; shoot meristem
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA clone of the previously unreported low-temperature-induced gene blt101 was isolated after a differential screen of a cDNA library prepared from low-temperature (6 °C day/2 °C night) grown barley shoot meristems. Southern blot analysis of barley ditelosomic addition lines was used to assign this single-copy gene to the long arm of chromosome 4. Analysis of steady-state levels of blt101 mRNA showed the induction of this transcript in shoot meristems upon transfer of barley (cv. Igri) plants from control (20 °C/15 °C) to low (6 °C/2 °C) temperature treatment. Further, the high level of this transcript is maintained at low temperatures but is reduced on transfer from low to control temperatures. The gene is not induced by drought or by foliar application of ABA. Analysis of segregating doubled haploid lines shows that there is no specific association of this gene with either spring/winter growth habit or frost hardiness. Examination of the spatial expression pattern revealed ubiquitous expression of blt101 in low-temperature (6 °C/2 °C) grown barley shoot meristems, mature leaves and roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021541
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    Differential, temporal and spatial expression of genes involved in storage oil and oleosin accumulation in developing rapeseed embryos: implications for the role of oleosins and the mechanisms of oil-body formation (1993)
    Springer
    Plant molecular biology 23 (1993), S. 1015-1027 
    Details
    ISSN: 1573-5028
    Keywords: embryos ; gene expression ; oleosin ; rapeseed
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The temporal and spatial expression of oleosin and Δ9-stearoyl-ACP desaturase genes and their products has been examined in developing embryos of rapeseed, Brassica napus L. var. Topas. Expression of oleosin and stearate desaturase genes was measured by in situ hybridisation at five different stages of development ranging from the torpedo stage to a mature-desiccating embryo. The temporal pattern of gene expression varied dramatically between the two classes of gene. Stearate desaturase gene expression was relatively high, even at the torpedo stage, whereas oleosin gene expression was barely detectable at this stage. By the stage of maximum embryo fresh weight, stearate desaturase gene expression had declined considerably while oleosin gene expression was at its height. In contrast to their differential temporal expression, the in situ labelling of both classes of embryo-specific gene showed similar, relatively uniform patterns of spatial expression throughout the embryo sections. Immunogold labelling of ultra-thin sections from radicle tissue with anti-oleosin antibodies showed similar patterns to sections from cotyledon tissue. However, whereas at least three oleosin isoforms were detectable on western blots of homogenates from cotyledons, only one isoform was found in radicles. This suggests that some of the oleosin isoforms may be expressed differentially in the various types of embryo tissue. The differential timing of stearate desaturase and oleosin gene expression was mirrored by similar differences in the timing of the accumulation of their ultimate products, i.e. storage oil and oleosin proteins. Oil-body fractions prepared from young (2.5 mg) embryos contained very little oleosin protein, as examined by SDS-PAGE and western blotting, whereas identically prepared fractions from dry seeds contained over 10% (w/w) oleosin. Dehydration of oil bodies from young embryos resulted in their breakdown and coalescence into large clumps of oil which could not be re-emulsified, even after rehydration. In contrast, the oleosin-rich oil bodies from mature embryos were stable to dehydration and subsequent rehydration. It is suggested that, in developing rapeseed embryos, the accumulation of storage oil and oleosins is not concomitant but that the eventual deposition of oleosins onto the surfaces of storage oil bodies is essential for their stability during seed desiccation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021816
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    Isolation and characterization of pollen-specific maize genes with sequence homology to ragweed allergens and pectate lyases (1993)
    Springer
    Plant molecular biology 23 (1993), S. 1061-1065 
    Details
    ISSN: 1573-5028
    Keywords: allergens ; gene expression ; microsporogenesis ; pectate lyase ; pollen ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA clone (Zm58.1) was isolated by differential screening from a cDNA library made to mature Zea mays pollen, and shown to be pollen-specific by RNA blot analysis. When this partial-length clone was used to probe a genomic library, a similar but distinct pollen-specific genomic clone (68% sequence identity) was isolated (Zm58.2). The putative proteins coded for by these two clones show sequence homology to several flower-expressed gene products from various plant species, including known pollen allergens from short ragweed (Ambrosia artemisiifolia), and to pectate lyases from the plant pathogenic bacteria Erwinia spp. The two genes map to different chromosomes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00021820
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    A novel blue light- and abscisic acid-inducible gene of Arabidopsis thaliana encoding an intrinsic membrane protein (1993)
    Springer
    Plant molecular biology 23 (1993), S. 1187-1198 
    Details
    ISSN: 1573-5028
    Keywords: Arabidopsis thaliana ; blue light ; floral induction ; gene expression ; membrane protein ; DNA sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Continuous irradiation with blue light (400–500 nm) induces flower formation in plantlets of Arabidopsis thaliana (C24) while red light (600–700 nm) is ineffective. This observation started a search for genes that are activated by blue light and initiate the morphogenic programme leading to flower formation. Several genes were identified via their cDNAs. From these clone AthH2, with an open reading frame for a hydrophobic 30.5 kDa polypeptide, was selected for further characterization of the corresponding gene. From a genomic library a DNA fragment of about 6.4 kb was isolated, comprising the coding region as well as 5′-upstream and 3′-downstream flanking segments. The coding region is composed of four exons, which specify a polypeptide of 286 amino acids. Several potential regulatory elements were found between position −670 and −1140 including GA and ABA sequence motifs. The latter could account for the observed induction of the AthH2 gene by ABA. Southern blot analysis of Arabidopsis genomic DNA suggests that the AthH2 gene is encoded by a single-copy gene. Hydropathy plots and secondary structure analysis of the putative polypeptide predict six membrane-spanning domains implicating a function as transmembrane channel protein. It displays significant homology with the proteins TR7a of pea (82%) and RD 28 of A. thaliana (68%).
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00042352
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    Plasmid retention and gene expression in suspended and biofilm cultures of recombinant Escherichia coli DH5α(pMJR1750) (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 41 (1993), S. 211-220 
    Details
    ISSN: 0006-3592
    Keywords: plasmid retention ; gene expression ; biofilm ; β-galactosidase ; segregational instability ; Escherichia coli ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Differences in plasmid retention and expression are studied in both suspended and biofilm cultures of Escherichia coli DH5α(PMJR1750). An alternative mathematical model is proposed which allows the determination of plasmid loss probability in both suspended batch and continuously fed biofilm cultures. In our experiments, the average probability of plasmid loss of E. coli DH5α(pMJR1750) is 0.0022 in batch culture in the absence of antibiotic selection pressure and inducer. Under the induction of 0.17 MM IPTG, the maximum growth rate of plasmid-bearing cells in suspended batch culture dropped from 0.45 h-1 to 0.35 h-1 and the β-galactosidase concentration reached an experimental maximum of 0.32. pg/cell 4 hours after the initiation of induction. At both 0.34 and 0.51 mM IPTG, growth rates in batch cultures decreased to 0.16 h-1, about 36% of that without IPTG, and the β-galactosidase concentration reached an experimental maximum of 0.47 pg/cell 3 hours after induction.In biofilm cultures, both plasmid-bearing and plasmid-free cells in increase with time reaching a plateau after 96 hours n the absence of both the inducer and any antibiotic selection pressure. Average probability of plasmid loss for biofilm-bound E. coli DH5β(pMJR1750) population was 0.017 without antibiotic selection. Once the inducer IPTG was added, the concentration of plasmid-bearing cells in biofilm dropped dramatically while plasmid-free cell numbers maintained unaffected. The β-galactosidase concentration reached a maximum in all biofilm experiments 24 hours after induction; they were 0.08, 0.1, and 0.12 pg/cel under 0.17, 0.34, and 0.51 mM IPTG, respectively. © 1993 John Wiley & Sons, Inc.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260410207
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    Regulation of olfactory neuron gene expression (1993)
    Springer
    Cytotechnology 11 (1993), S. 17-22 
    Details
    ISSN: 1573-0778
    Keywords: olfactory neuroepithelium ; neural development and regeneration ; neuronal phenotype ; gene expression ; OMP ; transgenic mice ; B50/GAP43 ; OLF-1 ; transcription
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00749053
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    Expression of thelacZ gene targeted to the HPRT locus in embryonic stem cells and their derivatives (1993)
    Springer
    Transgenic research 2 (1993), S. 1-13 
    Details
    ISSN: 1573-9368
    Keywords: gene expression ; lacZ ; HPRT ; ES cells ; gene targeting
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenes in mice often exhibit different expression patterns in different transgenic lines. While the basis for this phenomenon is not understood, it is widely believed that the site at which the transgene becomes integrated into the mouse genome is a major factor in determining the pattern of expression. Most transgenic mice have been produced by microinjection of DNA into the male pronucleus, which results in integration of tandem arrays of the transgene at random chromosomal sites. In the experiments described in this report, electroporation of embryonic stem (ES) cells was used to place single copies of alacZ transgene into either random sites or into the HPRT (hypoxanthine phosphoribosyl transferase) locus of the mouse genome. Expression oflacZ was assayed by histochemical staining forEscherichia coli β-galactosidase activity in ES cells and in differentiated derivatives obtained by teratocarcinoma formation. Several of the randomly integrated cell lines expressedlacZ at high levels in a variety of cell types present in the tumours, but most notably in epithelial cells. Targeted cell lines withlacZ in opposite orientation to the direction of HPRT gene transcription also expressed well in epithelial cells, but the targeted cell lines did not express in a wider variety of cell types than some of the nontargeted cell lines. Targeted cell lines transcribinglacZ in the same orientation as HPRT transcription did not express high levels oflacZ in any differentiated cell type. Analysis of transcripts suggested that this orientation effect may have been the result of transcriptional interference perpetrated by the HPRT gene promoter.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01977675
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    Molecular approaches to the study of evolution and phylogeny of the Nemertina (1993)
    Springer
    Hydrobiologia 266 (1993), S. 255-265 
    Details
    ISSN: 1573-5117
    Keywords: gene expression ; nucleotide sequence ; invertebrate
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Molecular biological tools currently available to us are revolutionizing the way in which we can address questions in evolutionary biology. The purpose of this article is to provide an overview of molecular techniques and applications available to biologists who are interested in evolutionary studies but who have little acquaintance with molecular biology. In evolutionary biology, techniques designed to determine degree of nucleic acid similarity are in common use and will be dealt with first. Another approach, namely gene expression studies, has strong implications for evolutionary biology but generally requires substantial familiarity with molecular biological tools. Expression studies provide powerful tools for discerning processes of speciation, as in the selection of genetic variants, as well as discerning lineages, e.g., expression of specific homeobox genes during segment formation. For investigations where either nucleic acid identity or gene expression are the ultimate goal, detailed information, protocols and appropriate controls are beyond the scope of this work but, where possible, recent review articles are cited.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00013373
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    The effect of thyroid hormones and 5-azacytidine on transcription of malic enzyme and 6-phosphogluconate dehydrogenase genes (1993)
    Springer
    Bulletin of experimental biology and medicine 116 (1993), S. 1376-1378 
    Details
    ISSN: 1573-8221
    Keywords: DNA methylase ; gene expression ; thyroid hormones ; 5-azacytidine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00805150
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    Inhibition of induced endochondral bone development in caffeine-treated rats (1993)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 52 (1993), S. 171-182 
    Details
    ISSN: 0730-2312
    Keywords: caffeine ; bone matrix implants ; delayed ossification ; osteoblasts ; gene expression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We have addressed questions raised by the observation in fetal rats of delayed ossification induced by caffeine at maternal doses above 80 mg/kg body weight per day. The effect of caffeine on endochondral bone development and mineralization has been studied in an experimental model system of bone formation which involves implantation of demineralized bone particles (DBP) in subcutaneous pockets of young growing rats. Caffeine's effects on cellular events associated with endochondral ossification were examined directly by quantitating cellular mRNA levels of chondrocyte and osteoblast growth and differentiation markers in DBP implants from caffeine-treated rats harvested at specific stages of development (day 7 through day 15). Oral caffeine administration to rats implanted with DBP resulted in a dose dependent inhibition of the formation of cartilage tissue in the implants. Histologic examination of the implants revealed a decrease in the number of cells which were transformed to chondrocytes compared to control implants. Those cartilaginous areas that did form, however, proceeded through the normal sequelae of calcified cartilage and bone formation. At the 100 mg/kg dose, cellular levels of mRNA for histone, collagen type II, and TGFβ were all reduced by greater than 40% of control implants consistent with the histological findings. Alkaline phosphatase activity in the implants and mRNA levels for proteins reflecting the hypertrophic chondrocyte and bone phenotype, collagen type I and osteocalcin were markedly decreased compared to controls. Lower doses of 50 and 12.5 mg/kg caffeine also resulted in decreased cellular proliferation and transformation to cartilage histologically and reflected by significant inhibition of type II collagen mRNA levels (day 7). The effects of caffeine on gene expression observed in vivo during the period of bone formation (day 11 to day 15) in the DBP model were similar to the inhibited expression of H4, alkaline phosphatase, osteocalcin, and osteopontin found in fetal rat calvarial derived osteoblast cultures following 24 hour exposure of the cultures to 0.4 mM caffeine. Thus the observed delayed mineralization in the fetal skeleton associated with caffeine appears to be related to an inhibition of endochondral bone formation at the early stages of proliferation of undifferentiated mesenchymal cells to cartilage specific cells as well as at later stages of bone formation.
    Additional Material: 9 Ill.
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    URL: http://dx.doi.org/10.1002/jcb.240520209
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    Activities of growth factors in preimplantation embryos (1993)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 53 (1993), S. 280-287 
    Details
    ISSN: 0730-2312
    Keywords: insulin-related factors ; epidermal growth factors ; cytokines ; gene expression ; growth factor receptors ; platelet-derived growth factors ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The development of the mammalian preimplantation embryo in vitro occurs more slowly and less successfully compared to development in the uterus. The fact that it can occur at all in a defined protein-free medium suggests that the process is autonomous. Accumulated evidence indicates that a number of peptide growth factors contribute in an autocrine fashion to preimplantation development. Other growth factors are maternally derived and act in a paracrine manner on the embryo. Some of these factors such as insulin-related factors stimulate growth preferentially, but others such as epidermal growth factor (EGF) play more important roles in differentiation. Several cytokines appear to be implicated in peri-implantation events and in maternal-fetal interactions. At this stage, the data are mostly descriptive. Are all these different growth factors and receptors necessary for early development? Some implications of apparent redundancy of gene expression are discussed and future studies are predicted.
    Additional Material: 3 Tab.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240530403
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    Expression of histone and alkaline phosphatase genes in UMR 106-01 rat osteoblast-like cells exposed to the hoechst dye H33342 (1993)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 51 (1993), S. 175-180 
    Details
    ISSN: 0730-2312
    Keywords: fluorescent dye H33342 ; histone ; gene expression ; cell cycle ; osteoblast ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The fluorescent Dye H33342 (H342) is a bis-benzimidazole used for intravital fluorescent staining. In this report, we found that H342 completely abolished histone 2a mRNA but had no effect on alkaline phosphatase gene expression and protein synthesis in UMR 106-01 rat osteoblast-like cells. The complete loss of histone 2a mRNA occurred after only 20 min of treatment with H342. This effect is unlikely to be a result of inhibition of DNA synthesis, which was only partly suppressed. The mechanism of the action of H342 on histone 2a mRNA is presently unknown. © 1993 Wiley-Liss, Inc.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240510209
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    1α,25-Dihydroxyvitamin D3-induced changes in intracellular pH in osteoblast-like cells modulate gene expression (1993)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 53 (1993), S. 234-239 
    Details
    ISSN: 0730-2312
    Keywords: 1,25-Dihydroxyvitamin D3 osteoblasts ; intracellular pH ; gene expression ; mRNA ; cell calcium ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: 1α,25-Dihydroxyvitamin D3 exerts rapid nongenomic effects on rat osteoblast-like cells independent of the classic nuclear receptor. These effects include changes in phospholipid metabolism and cell calcium. Intracellular calcium itself has been proposed to regulate intracellular pH in osteoblast cell lines. The purpose of this study was to determine the effect of 1α,25-dihydroxyvitamin D3 on intracellular pH, the relationship of changes in calcium to changes in pH, and the role of pH changes in genomic activation. 1α,25-Dihydroxyvitamin D3 increased intracellular pH within 10 min in rat osteoblast-like cells, an effect that was inhibited by removal of extracellular sodium and by the biologically inactive epimer 1β,25-dihydroxyvitamin D3. The hormone increased intracellular calcium in Quin 2 loaded cells in the presence and absence of extracellular sodium. The 1α,25-dihydroxyvitamin D3-induced increments in osteocalcin and osteopontin mRNA levels were abolished in sodium-free medium. The results indicate that 1α,25-dihydroxyvitamin D3-induced increments in cellular calcium precede cell alkalinization and that these changes in intracellular pH may modulate steady-state mRNA levels of genes induced by vitamin D.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240530308
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    Cytokine regulation of metalloproteinase gene expression (1993)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 53 (1993), S. 288-295 
    Details
    ISSN: 0730-2312
    Keywords: metalloproteinases ; cytokines ; gene expression ; extracellular matrix ; growth factors ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Matrix metalloproteinases belong to a family of zinc-dependent enzymes capable of degrading extracellular matrix and basement membrane components. Their expression is greatly modulated by cytokines and growth factors and involves the gene products of the Fos and Jun families of oncogenes. After extra(peri)cellular activation, their activity can be further controlled by specific tissue inhibitors of metalloproteinases. A correct balance between these regulatory mechanisms is necessary to ensure matrix remodeling in normal physiological processes such as embryonic development, but the overexpression of these enzymes may initiate or contribute to pathological situations such as cartilage degradation in rheumatoid arthritis or to tumor progression and metastasis. Delineation of the mechanisms of metalloproteinase and metalloproteinase inhibitors gene expression, understanding of their mode of interactions, and characterization of their patterns of expression in various tissues in normal and pathological states will lead to new therapeutic strategies to counteract the deleterious effects of matrix metalloproteinases in human disease.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240530404
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    Estrogen receptor-negative breast cancer cells transfected with the estrogen receptor exhibit increased RARα gene expression and sensitivity to growth inhibition by retinoic acid (1993)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 53 (1993), S. 394-404 
    Details
    ISSN: 0730-2312
    Keywords: transfection ; CAT assays ; gene expression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: We and others have shown previously that retinoic acid (RA) selectively inhibits the growth of estrogen receptor (ER)-positive human breast carcinoma (HBC) cells and ER-negative cells are refractory to RA inhibition of growth. The ER-negative cells inherently express lower levels of RARα and retinoic acid response element (RARE)-mediated RA-induced CAT activity. In this study we report that when ER-negative MDA-MB-231 cells were transfected with the ER gene they not only expressed higher levels of RARα and RARE-mediated RA-induced CAT gene expression, but their growth was now inhibited by RA. Estrogen enhanced RARα gene expression not only in established ER-positive cell lines but also in ER-transfected MDA-MB-231 cells. The estrogen effect appears to be direct and at the gene transcription level since it did not alter the stability of RARα mRNA and cycloheximide failed to block estrogen-mediated enhancement of RARα gene expression. Our data strongly suggest that ER-mediated enhancement of RARα levels plays an important role in RA inhibition of HBC growth. In addition, we also report here that HBC cells appear to express a unique isoform(s) of RARα which was detected only when the full-length RARα cDNA was used as a probe; the RARα1 and RARα2 specific probes failed to hybridize with the HBC specific RARα message.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240530417
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  • 138
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    Expression of the glucose oxidase gene from Aspergillus niger in Hansenula polymorpha and its use as a reporter gene to isolate regulatory mutations (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 9 (1993), S. 625-635 
    Details
    ISSN: 0749-503X
    Keywords: Aspergillus niger ; gene expression ; glucose oxidase ; Hansenula polymorpha ; MF-α secretion leader sequence ; methylotrophic yeast ; methanol oxidase promoter ; recombinant protein ; regulatory mutants ; reporter gene ; super-secretion mutant ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The glucose oxidase gene (god) from Aspergillus niger was expressed in Hansenula polymorpha using the methanol oxidase promoter and transcription termination region and the MF-α leader sequence from Saccharomyces cerevisiae to direct secretion. The expression cassette was cloned into the S. cerevisiae vector YEp13 and used to transform H. polymorpha strain A16. In the initial transformants plasmid replication was unstable, but was stabilized by a growth regime consisting of alternating cycles of selective and non-selective growth. The stabilized strain was grown to high cell density by fed-batch fermentation. Upon induction of the MOX promoter, glucose oxidase synthesis was initiated. At the end of the fermentation, the culture density was 76 g dry weight/1 and 108 IU/ml (0·5 g/l or 0·65% dry weight) glucose oxidase was found in the culture medium; a further 86 IU/ml (0·43 g/l or 0·56% dry weight) was recovered from the cell lysate. A plate assay was used to monitor glucose oxidase levels in individual colonies. This was then used to isolate mutants which showed abnormal regulation of god expression or which showed an altered pattern of secretion. One mutant, which showed increased production of glucose oxidase, was grown to high cell density by fed-batch fermentation (100·6 g/l) and produced 445 IU/ml (2·25 g/l or 2·2% dry weight) extracellularly and 76 IU/ml (0·38 g/l or 0·4% dry weight) intracellularly. The mutant thus not only increased total production but exported 83% of the total enzyme made compared to 55% in the parent strain.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320090609
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  • 139
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    Lack of concordance between heat shock proteins and the development of tolerance to teratogen-induced neural tube defects (1993)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 14 (1993), S. 137-147 
    Details
    ISSN: 0192-253X
    Keywords: Heat-shock proteins ; teratogenicity, tolerance and cross-tolerance ; neural tube defects ; gene expression ; In situ transcription ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The present study was undertaken to examine the role of heat shock response in the development of tolerance and cross-tolerance in an in vivo murine model of teratogen-induced neural tube defects. The experimental paradigm designed to address this question was to utilize inbred mouse strains that differed in their sensitivity to hyperthermia and valproic acid induced neural tube defects, subjecting the dams to subteratogenic pretreatments with either heat or valproic acid at two different timepoints during development prior to the administration of the teratogenic insult. A statistically significant reduction in the frequency of neural tube defects and/or embryolethality following a pretreatment in dams subsequently exposed to a teratogenic treatment was considered evidence for the induction of tolerance. This was observed in the SWV embryos exposed to the 38°C pretreatment at 8:06 and to embryos exposed to either pretreatment temperature at 8:10 priorto a teratogenic heat shock at 8:12. In the LM/Bc embryos, only the 41°C pretreatment at 8:06 induced thermotolerance. There was no evidence of tolerance induced in either mouse strain using valproic acid. On the other hand, cross-tolerance was clearly demonstrated in this study, with a low temperature (41°C) pretreatment successfully protecting SWV fetuses from a subsequent teratogenic treatment with valproic acid, while valproic acid (200 mg/kg) was effective in reducing the risk of hyperthermia-induced neural tube defects in the LM/Bc fetuses. In all instances, tolerance was induced in the absence of significant induction of hsp synthesis. The lack ofconcordance between hsps and thermotolerance suggests that some other factor(s) is involved in conferring thermotolerance on developing murine embryos. © 1993 Wiley-Liss, Inc.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020140208
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    Rapid induction and clearance of TGFβ1 is an early response to wounding in the mouse embryo (1993)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 14 (1993), S. 225-238 
    Details
    ISSN: 0192-253X
    Keywords: Growth factor ; wound healing ; embryo ; in situ hybridisation ; immunohistochemistry ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The TGFβ family of growth factors has been implicated as playing a significant role in many aspects of embryonic morphogenesis, and also as a mediator of adult tissue repair processes. Unlike the situation in the adult, rissue repair in the embryo does not result in scarring, and it has been suggested that this might be due, in part, to reduced levels of growth factors, particularly TGFβ, at the wound site. We have examined the expression patterns of TGFβ genes following wounding of limb bud lesions in cultured Ell.5 mouse embryos. The timetable of wound closure was investigated by standard light and electron microscopy from the time of wounding until the lesion had re-epithelialised 24 hours later. The expression of transcripts for each of the three TGFβ genes was examined at various time points during the healing process using radioactive in situ hybridisation to tissue sections and wholemount non-radioactive in situ hybridisation to embryo pieces. Within l to 3 hours of wounding, transcripts encoding TGFβl were rapidly induced within the epithelial cells of the wound margin, particularly those cells at the ventral aspect of the wound. By 3 to 6 hours post-wounding, TGFβl transcripts were detectable in the mesenchyme of the wound bed. No TGFβS induction was observed, and possible TGFβ2 induction was largely obscured by endogenous expression associated with pre-cartilage mesenchymal condensation. Immunocytochemical analysis of tissue sections of the wound demonstrated a rapid induction of TGFβl protein within l hour post-wounding, but also a subsequent rapid clearance of the protein from the wound site such that, by 18 hours post-wounding, TGFβl levels had returned to near background. These data are discussed in terms of the molecular mechanisms underlying embryonic wound healing and the significance of the results to an understanding of scarring following adult tissue repair. © 1993Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020140309
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    Activity of a microinjected inducible murine hsp68 gene promoter depends on plasmid configuration and the presence of heat shock elements in mouse dictyate oocytes but not in two-cell embryos (1993)
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 14 (1993), S. 92-102 
    Details
    ISSN: 0192-253X
    Keywords: Heat shock promoter ; mouse preim-plantation embryos ; mouse dictyate oocytes ; gene expression ; microinjection ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: After fertilization in the mouse, the zygotic genome is activated in two-cell embryos by the spontaneous expression, among other genes, of the major inducible heat shock gene, hsp68, in the absence of heat-inducibility of heat shock genes. To obtain information on this phenomenon, we have probed one- and two-cell embryo's ability to express microinjected reporter DNA constructs, containing the Escherichia coli lacZ gene driven by promoters from early SV40 genes, the human ß-actin gene, and the normal or HSE-deleted mouse hsp68 gene. Activity of these promoters was also tested in mouse granulosa cells and dictyate oocytes, as a function of circular/linear construct configuration and occurrence of heat shock. The hsp68 promoter was heat-inducible in both granulosa cells and oocytes. Its heat activation required the presence of HSEs and, in the oocytes, of construct linear configuration. In the embryos however, this promoter was expressed in dependently of the presence of HSEs and of construct configuration, and its activity was not affected by heat shock. When constructs with early SV40 and ß-actin promoters were injected into one-cell embryos, they appeared to be inactivated with the first embryonic cleavage, in agreement with previous observations [Wiekowski et ai., 1992]. By contrast, both normal and HSE-deleted hsp68 promoters maintained their activity through the first cleavage, providing the first evidence of a gene escaping such transcriptional repression. Present results confirm previous findings on hsp68 expression during early mouse development, and suggest that this activation is mediated by a factor(s) other than HSF. © 1993Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/dvg.1020140203
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    Expression of hornet genes encoding venom allergen antigen 5 in insects (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 22 (1993), S. 303-313 
    Details
    ISSN: 0739-4462
    Keywords: baculovirus ; Autographa californica nuclear polyhedrosis virus ; insect toxin ; protein production ; gene expression ; hornet venom ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Antigen 5, also known as Dol m V, is a major allergen found in the venom of the baldfaced hornet, Dolichovespula maculata. We have inserted the f10 and f17 cDNAs, which encode hornet antigen 5 (HA5) forms 2 (Dol m; VB) and 3 (Dol m VA), respectively, into the genome of the baculovirus, AcMNPV, to produce the recombinant baculovirus gene expression vectors, vEV-HA5f10 and vEV-HA5f17. Insect cells infected with either vEV-HA5f10 or vEV-HA5f17 produce and secrete a novel protein with an electrophoretic mobility which is similar if not identical to authentic mature Dol m V. The gene products also react specifically with a polyclonal antiserum raised to Dol m VB as expected. Dol m V gene products were not acutely toxic when injected into insect larvae. However, infection of fifth instar larvae with vEV-HA5f17 resulted in premature melanization of the larvae and lower weight gain than infection with control virus. Thus, the Dol m V gene product has a subtle, possibly cytotoxic or biochemical effect on insects. The expression systems may prove useful in further structural and functional characterization of these proteins. © 1993 Wiley-Liss, Inc.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/arch.940220302
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    Insecticidal potential of the insect parvovirus GmDNV (1993)
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 22 (1993), S. 345-356 
    Details
    ISSN: 0739-4462
    Keywords: densovirus ; larvacidal activity ; gene expression ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The insecticidal potential of Galleria mellonella densovirus (GmDNV) in third, fourth, and fifth instar larvae of the host, the greater wax moth, as a step toward the construction of a molecular vector for the introduction and expression of foreign genes in the larvae of these insects was studied. Third instar larvae are most susceptible to GmDNV. Viral RNA synthesis is more rapid in this stage and slowest in the fifth instar. Infection of prepupae by intradermic injection or by horzontal spread inhibited pupation. GmDNV DNA is also infectious when introduced as a calcium phosphate precipitate. The two putative viral promoters were shown to be capable of driving the expression of the reporter gene chloramphenicol acetyltransferase (CAT) in DNA-injected larvae. © 1993 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/arch.940220304
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  • 144
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    Expression of an islet regenerating (reg) gene in isolated rat islets: effects of nutrient and non-nutrient growth factors (1992)
    Springer
    Diabetologia 35 (1992), S. 238-242 
    Details
    ISSN: 1432-0428
    Keywords: Islets ; growth factors ; replication ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The expression of a novel regenerating (reg) gene has been reported previously in the regenerating islets of a surgical model of diabetes in rats. We exposed collagenase-isolated rat islets for three days to nutrient and non-nutrient growth factors in minimally supplemented RPMI medium (2.7 mmol/l glucose, 2% fetal calf serum), and investigated the relationship between reg gene expression and islet cell replication. RNA was prepared from half of the islets by homogenisation in guanidinium isothiocyanate followed by phenol/chloroform extraction. Northern/dot blot analyses were used to semi-quantify reg mRNA. Islet cell replication was estimated by culturing the remaining islets in radiolabelled thymidine to determine de novo DNA synthesis. Thymidine uptake was stimulated by the following factors: 11 mmol/l glucose (50% increase); 10% amino acids (126% increase); 10% fetal calf serum (39% increase); 100 ng/ml insulin (45% increase); 250 ng/ml growth hormone (65% increase); 1.5 nmol/l aldosterone (29% increase); 2U/ml platelet derived growth factor (116% increase). The results are expressed as a percentage of the thymidine incorporated into control islets cultured in minimal RPMI (1118±100 (SD) cpm/μg protein, n=15). Increased islet cell replication was paralleled in each case by a clear rise in reg mRNA expression compared to controls. Furthermore, the rank order for reg gene expression was the same as that for thymidine uptake (r=0.90). The present findings suggest a clear association between reg gene expression and islet cell replication in vitro, and are the first to demonstrate reg gene expression in response to individual growth factors.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00400923
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    Overexpression and purification of enzymatically active recombinant integrase protein of rous sarcoma virus (1992)
    Springer
    Virus genes 6 (1992), S. 301-306 
    Details
    ISSN: 1572-994X
    Keywords: molecular cloning ; gene expression ; integrase protein ; enzyme purification ; Rous sarcoma virus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The carboxy-terminal domain of polymerase gene of Rous sarcoma virus was cloned into an expression vector under the control oflac regulatory elements, resulting in the plasmid pMF1413. Upon isopropyl-β-D-thiogalactopyranoside induction, viral integration (IN) protein was expressed in large quantity inEscherichia coli. The expressed recombinant protein was prepurified by successive washing of the bacterial pellet with 0.1 M NaCl and detergents. Further purification was performed in high yield by standard chromatography methods. The purified enzyme revealed selective DNA cleaving activity on supercoiled plasmid with the LTR-LTR junction fragment. The reaction was metal ion dependent, with a preference for Mn2+ over Mg2+, and showed substrate specificity at 1 mM MnCl2.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01702568
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    Comparative sequence analysis and expression of bovine PrP gene in mouse L-929 cells (1992)
    Springer
    Virus genes 6 (1992), S. 343-356 
    Details
    ISSN: 1572-994X
    Keywords: Bovine PrP ; cDNA ; nucleotide sequence ; gene expression ; transfection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract A cDNA clone encoding bovine scrapie-associated fibril protein, PrP, from a bovine brain cDNA library and six amplified genomic DNA clones of bovine PrP were characterized. These clones possessed specific characteristics observed in other animal PrP genes. However, the bovine PrP was divided into two types by the number of repeats. One possessed four octapeptide repetitive sequences, like other animal PrP genes, and consisted of 256 amino acids; the other had five such repetitive sequences and 264 amino acids. The amino acid sequence of the former bovine PrP agreed with that of sheep PrP up to the 165th amino acid from the N-terminus. Bovine PrP cDNA introduced into mouse L-929 cells were stably expressed. The expression level of recombinant bovine PrP in the cells judged by immunofluorescence was higher than that of authentic mouse PrP. The recombinant PrP comigrated with authentic bovine PrP in SDS-polyacrylamide gel electrophoresis, suggesting that the recombinant product was fully glycosylated in L-929 cells. Distinct bundles of the intermediate filaments were frequently seen at the perinuclear region of the cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01703083
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    Transient expression assay in a baculovirus system using firefly luciferase gene as a reporter (1992)
    Springer
    Virus genes 6 (1992), S. 379-386 
    Details
    ISSN: 1572-994X
    Keywords: recombinant DNA ; gene expression ; polyhedrin gene promoter ; expression vector ; DNA sequencing
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Transient gene expression assays were developed to assess the function of the regulatory sequences of baculovirusesBombyx mori nuclear polyhedrosis virus (BmNPV) andAutographa californica nuclear polyhedrosis virus (AcNPV) in insect cells ofBombyx mori andSpodoptera frugiperda, respectively. DNA sequences encoding luciferase (luc) of the fireflyPhotinus pyralis was successfully employed in the expression assay as a reporter gene. Recombinant plasmids were constructed containing the luc gene under control of baculovirus-specific or heterologous promoters. Cotransfection ofBombyx mori andSpodoptera frugiperda cells with recombinant plasmids carrying virus-specific promoter sequences and BmNPV and AcNPV DNA, respectively, gave rise to efficient synthesis of luciferase (Luc), while heterologous promoters induced a low level of luc expression. We found that flanking sequences of the AcNPV DNA in the transfer plasmid contained an unknown promoter conferring an efficient luc expression. The activity of this promoter was modulated by the polh promoter sequences. The assay allows one to conduct highly sensitive monitoring of the transient expression of foreign genes from the transfecting plasmids prior to construction of recombinant viruses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01703086
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  • 148
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    Molecular events at nematode-induced feeding sites (1992)
    Springer
    European journal of plant pathology 98 (1992), S. 175-181 
    Details
    ISSN: 1573-8469
    Keywords: Nematodes ; Globodera ; plant pathogen ; infection ; monoclonal antibodies ; PCR ; cDNA libraries ; gene expression ; modified plant cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Current control of nematodes is inadequate and this justifies work towards the design of novel bases for plant defence. Our approach for cyst nematodes began by improving understanding of critical events in the establishment of these biotrophic pathogens. The first step involved development of an experimental system for achieving synchronous infection and establishment of cyst-nematodes in roots. Monoclonal antibodies have been raised against these nematodes, their specificity defined and those of particular interest used to define events in the establishment of the animals within plants. A similar approach has been explored for host responses using antibodies raised to plant tissue containing feeding sites. Changes in translatable mRNA populations at the feeding site have been described.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01974484
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    Molecular cloning, nucleotide sequence and expression of a cDNA encoding an intracellular protein tyrosine phosphatase, PTPase-2, from mouse testis and T-cells (1992)
    Springer
    Molecular and cellular biochemistry 118 (1992), S. 91-98 
    Details
    ISSN: 1573-4919
    Keywords: mouse ; protein tyrosine phosphatase ; cDNA cloning ; nucleotide sequence ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The PTP-2 cDNA encoding an intracellular protein tyrosine phosphatase (PTPase-2) was isolated and sequenced from mouse testis and T-cell cDNA libraries. This PTP-2 cDNA was found to be homologous to human PTP-TC and rat PTP-S, and contained 1,551 nucleotides, including 1,146 nucleotides encoding 382 amino acids as well as 5′ (61 nucleotides) and 3′ (344 nucleotides) non-coding regions. Northern blot analysis indicated that PTP-2 mRNA of 1.9 Kb was most abundant in testis and kidney, although it was also present in spleen, muscle, liver, heart and brain.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00249698
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  • 150
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    Active cell death in hormone-dependent tissues (1992)
    Springer
    Cancer and metastasis reviews 11 (1992), S. 197-220 
    Details
    ISSN: 1573-7233
    Keywords: apotosis ; prostate ; breast ; hormones ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Active cell death (ACD) in hormone-dependent tissues such as the prostate and mammary gland is readily induced by hormone ablation and by treatment with anti-androgens or anti-estrogens, calcium channel agonists and TGFβ. These agents induce a variety of genes within the hormone-dependent epithelial cells including TRPM-2, transglutaminase, poly(ADP-ribose) polymerase, Hsp27 and several other unidentified genes. Not all epithelial cells in the glands are equally sensitive to the induction of ACD. In the prostate, the secretory epithelial cells that are sensitive to hormone ablation are localized in the distal region of the prostatic ducts, and are in direct contact with the neighboring stroma. In contrast, the epithelial cells in the proximal regions of the ducts are more resistant to hormone ablation, probably because the permissive effects of the stroma are attenuated by the presence of the basal epithelial cells, which are intercalated between the epithelium and stroma. The underlying biology of ACD in prostate and mamary glands, and its relevance to hormone resistance, is discussed in this review.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00048064
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    Genetic approaches to the study of mitochondrial biogenesis in yeast (1992)
    Springer
    Antonie van Leeuwenhoek 62 (1992), S. 131-153 
    Details
    ISSN: 1572-9699
    Keywords: mitochondrial DNA ; mutational analysis ; nucleo-mitochondrial interactions ; gene expression ; membrane assembly ; respiratory deficiency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In contrast to most other organisms, the yeastSaccharomyces cerevisiae can survive without functional mitochondria. This ability has been exploited in genetic approaches to the study of mitochondrial biogenesis. In the last two decades, mitochondrial genetics have made major contributions to the identification of genes on the mitochondrial genome, the mapping of these genes and the establishment of structure-function relationships in the products they encode. In parallel, more than 200 complementation groups, corresponding to as many nuclear genes necessary for mitochondrial function or biogenesis have been described. Many of the latter are required for post-transcriptional events in mitochondrial gene expression, including the processing of mitochondrial pre-RNAs, the translation of mitochondrial mRNAs, or the assembly of mitochondrial translation products into the membrane. The aim of this review is to describe the genetic approaches used to unravel the intricacies of mitochondrial biogenesis and to summarize recent insights gained from their application.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00584467
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    The environment, microbes and bioremediation: microbial activities modulated by the environment (1992)
    Springer
    Biodegradation 3 (1992), S. 125-135 
    Details
    ISSN: 1572-9729
    Keywords: natural evolution ; directed evolution ; biodegradation ; environmental pollutants ; environmental signal transduction ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Microorganisms in nature are largely responsible for the biodegradation and removal of toxic and non-toxic chemicals. Many organisms are also known to have specific ecological niches for proliferation and colonization. The nature of the environment dictates to a large extent the biodegradability of synthetic compounds by modulating the evolutionary processes in microorganisms for new degradative genes. Similarly, environmental factors often determine the extent of microbial gene expression by activating or repressing specific gene or sets of genes through a sensory signal transduction process. Understanding how the environment modulates microbial activity is critical for successful bioremediative applications.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00129078
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    Biotechnology in the degradation and utilization of lignocellulose (1992)
    Springer
    Biodegradation 3 (1992), S. 219-238 
    Details
    ISSN: 1572-9729
    Keywords: cellulase ; gene expression ; lignin ; Phanerochaete chrysosporium ; Streptomyces cyaneus ; xylanase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Energy, Environment Protection, Nuclear Power Engineering , Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Lignocellulose is the predominant renewable resource. It uses include fuel, as the feedstock for the pulp and paper industry, and for animal nutrition. It also constitutes a large proportion of agricultural and urban waste. Biotechnology has roles in its efficient production and utilisation. The types of lignin substrates available for study of lignin biodegradation are described. The white rot fungus Phanerochaete chrysosporium is the archetypal system for the study of lignocellulose degradation, since it mineralises lignin and degrades both cellulose and hemicellulose. The salient features of the P. chrysosporium system are described. The lignin peroxidases are a family of proteins, and it is shown that expression of their genes is differential. P. chrysosporium is heterokaryotic with two gene equivalents that have abundant RFLPs. A set of basidiospore-derived strains with genetic compositions defined by such RFLPs provided the potential basis for a strain improvement programme for lignin degradation. However, analysis of this system using radiolabelled synthetic lignin (DHP) as the substrate confirmed previous evidence that both the substrate and the fungal cultures displayed much variation, so that it was difficult to quantify performance for this property. The cellobiohydrolase I enzymes are also coded for by a family of genes, and evidence is also presented for allelic variants, for differential expression and for differential splicing. In contrast, the cellobiohydrolase II function is encoded at a unique genetic locus. Approaches to an homologous integrative transformation system are discussed. Some actinomycete bacteria represent an alternative system for lignin solubilisation in which strains differ in their spectra of activities on lignocellulose substrates. The xylanase system of Streptomyces cyaneus is shown to include three enzymes, two of which are inducible by xylan. A novel assay method was developed and used to demonstrate that the third is constitutive and also non-repressible by glucose. It is proposed that this acts as a sensor for xylans in the environment that can yield breakdown products that are taken up and can then act as inducers of the other two enzymes. The studies on microbial lignocellulose degradation from different laboratories have allowed the formulation of specific biotechnological goals, and some of the problems and opportunities in this area are identified.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00129085
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  • 154
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    Peroxisome biogenesis inSaccharomyces cerevisiae (1992)
    Springer
    Antonie van Leeuwenhoek 62 (1992), S. 63-78 
    Details
    ISSN: 1572-9699
    Keywords: biogenesis ; gene expression ; mutants ; peroxisomes ; yeast
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The observation that peroxisomes ofSaccharomyces cerevisiae can be induced by oleic acid has opened the possibility to investigate the biogenesis of these organelles in a biochemically and genetically well characterized organism. Only few enzymes have been identified as peroxisomal proteins inSaccharomyces cerevisiae so far; the three enzymes involved in β-oxidation of fatty acids, enzymes of the glyoxylate cycle, catalase A and the PAS3 gene product have been unequivocally assigned to the peroxisomal compartment. However, more proteins are expected to be constituents of the peroxisomes inSaccharomyces cerevisiae. Mutagenesis ofSaccharomyces cerevisiae cells gave rise to mutants unable to use oleic acid as sole carbon source. These mutants could be divided in two groups: those with defects in structural genes of β-oxidation enzymes (fox-mutants) and those with defects in peroxisomal assembly (pas-mutants). All fox-mutants possess morphologically normal peroxisomes and can be assigned to one of three complementation groups (FOX1, 2, 3). All three FOX genes have been cloned and characterized. The pas-mutants isolated are distributed among 13 complementation groups and represent 3 different classes: peroxisomes are either morphologically not detectable (type I) or present but non-proliferating (type II). Mislocalization concerns all peroxisomal proteins in cells of these two classes. The third class of mutants contains peroxisomes normal in size and number, however, distinct peroxisomal matrix proteins are mislocalized (type III). Five additional complementation groups were found in the laboratory of H.F. Tabak. Not all PAS genes have been cloned and characterized so far, and only for few of them the function could be deduced from sequence comparisons. Proliferation of microbodies is repressed by glucose, derepressed by non-fermentable carbon sources and fully induced by oleic acid. The regulation of four genes encoding peroxisomal proteins (PAS1, CTA1, FOX2, FOX3) occurs on the transcriptional level and reflects the morphological observations: repression by glucose and induction by oleic acid. Moreover, trans-acting factors like ADR1, SNF1 and SNF4, all involved in derepression of various cellular processes, have been demonstrated to affect transcriptional regulation of genes encoding peroxisomal proteins. The peroxisomal import machinery seems to be conserved between different organisms as indicated by import of heterologous proteins into microbodies of different host cells. In addition, many peroxisomal proteins contain C-terminal targeting signals. However, more than one import route into peroxisomes does exist. Dissection of the import mechanism in a genetically well suited organism likeSaccharomyces cerevisiae together with further characterization and functional assignment of the PAS gene products will provide insight into the biogenesis of peroxisomes. Moreover, these studies will lead to a good model system for elucidation of the mechanisms underlying human peroxisomal disorders.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00584463
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  • 155
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    Phosphoribulokinase from ice plant: Transcription, transcripts and protein expression during environmental stress (1992)
    Springer
    Photosynthesis research 31 (1992), S. 127-138 
    Details
    ISSN: 1573-5079
    Keywords: environmental stress ; Mesembryanthemum crystallinum ; phosphoribulokinase ; gene expression ; protein expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The expression of PRK (phosphoribulokinase, E.C.2.7.1.19) in ice plant (Mesembryanthemum crystallinum) during development and under environmental stress was studied. cDNA clones were isolated and full-length cDNAs were characterized. Ice plant PRK is contained in a 1520 nucleotide transcript including a 126 nucleotide leader sequence, a 175 nucleotide 3′-end and a 20–30 nucleotide polyA+-stretch. The coding region, 397 codons, specifies a protein of Mr 44 064. The mature sequence is preceded by a transit peptide of approximately 46 amino acids. The mature portion of ice plant PRK is 86.4% identical to that of spinach and, e.g., 16.2% identical to PRK from Xanthomonas flavus. Under salt stress or cold adaptation conditions, the amount of mRNA declined by a factor of approximately three within days, followed by an increase to approximately pre-stress levels. The fluctuation in mRNA amount is not reflected on the level of transcription of the gene, suggesting post-transcriptional control, nor is PRK protein amount affected significantly over the short stress period. The recovery of transcript levels for photosynthesis-related proteins after stress appears to be a general response to environmental stresses that affect water status in ice plant. We suggest that the photosynthetic machinery in this facultative halophyte is effectively buffered from damage caused by such environmental stress.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028789
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  • 156
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    Respiratory metabolism and gene expression during seed germination (1992)
    Springer
    Plant growth regulation 11 (1992), S. 211-224 
    Details
    ISSN: 1573-5087
    Keywords: Germination ; energy metabolism ; gene expression ; regulation ; respiration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Oxygen uptake and carbon dioxide release rapidly increase in seeds during imbibition. The oxygen uptake is associated with oxidative phosphorylation through cytochrome oxidase. During the early stage of germination substrate level phosphorylation may also contribute to ATP production. All indications suggest that this route of ATP production is insignificant during aerobic germination. However, during oxygen stress, substrate level phosphorylation does significantly contribute to ATP production in some species. Carbohydrate oxidation plays a significant role in the germination process. Up to two thirds of the carbon from carbohydrate breakdown enters the tricarboxylic acid cycle through the phosphoenolpyruvate carboxylase reaction. This anapleurotic input into the Krebs cycle most probably reflects the high demand on intermediates from the cycle for biosynthesis. The extent to which other substrates are utilized for respiration is uncertain. Information regarding the levels of key metabolites and enzymes, as well as their cellular distribution is limited. The involvement of gene expression in the regulation of respiratory metabolism is poorly characterised. Several genes which have been cloned are only expressed during germination. With the exception of the early methionine labeled polypeptide, little is known about the function of these genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00024560
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  • 157
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    Effect of deletions 5′ to the translation initiation sequence on the expression of an mRNA in animal cells (1992)
    Springer
    Molecular biology reports 16 (1992), S. 277-284 
    Details
    ISSN: 1573-4978
    Keywords: translational initiation ; 18S rRNA ; mRNA secondary structure ; gene expression ; initiation mutants ; β-galactosidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To learn if an mRNA·18S rRNA interaction or a special secondary structure in the mRNA start region is essential for translation in eukaryotic cells, we constructed recombinant plasmids with the SV40 early promoter 5′ to part of the Escherichia coli tuf B-lacZ gene. Deletion of bases potentially complementary to the 18S rRNA highly increased the transient β-galactosidase expressed in transfected CHO cells. Deletion of bases that fostered formation of potential hairpins with the mRNA 5′-terminus or altered the structure of the coding region reduced β-galactosidase activity suggesting that these features of the mRNA secondary structure may be essential for initiation of translation. Computer aided analysis of the potential structure of 290 mRNAs suggests these are conserved features of the initiation region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419668
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  • 158
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    The early genetic response to light in the green unicellular alga Chlamydomonas eugametos grown under light/dark cycles involves genes that represent direct responses to light and photosynthesis (1992)
    Springer
    Plant molecular biology 18 (1992), S. 429-445 
    Details
    ISSN: 1573-5028
    Keywords: Chlamydomonas eugametos ; chlorophyll a/b-binding proteins ; circadian oscillator ; gene expression ; light-regulated genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In the green unicellular alga Chlamydomonas eugametos, cellular division is readily synchronized by light/dark cycles. Under these conditions, light initiates photosynthetic growth in daughter cells and begins the G1 phase. Genes whose expression is regulated upon illumination are likely to be important mechanisms controlling cell proliferation. To identify some of those genes, two cDNA libraries were prepared with poly(A)+ extracted from cells either stimulated with light for 1 h or held in darkness (quiescent cells) during the same period. To restrict our analysis to those genes that are part of the primary response, cells were incubated in presence of cycloheximide. Differential screening of approximately 40 000 clones in each library revealed 44 clones which hybridize preferentially with a [32P] cDNA probe derived from RNA of light-stimulated cells and 15 clones which react selectively with a [32P] cDNA probe synthesized from poly(A)+ RNA of quiescent cells. Cross-hybridization of these clones identified 4 independent sequences in the light-induced (LI) collection and 2 in the uninduced (LR) library. Four of these cDNAs correspond to mRNAs that are positively or negatively regulated upon activation of photosynthesis. One clone represents a mRNA that accumulates transitorily at both transitions. Finally, LI818 cDNA identifies a new chlorophyll a/b-binding (cab) gene family whose mRNA accumulation is controlled by light and a circadian oscillator. The endogenous timing system controls LI818 mRNA accumulation so that it precedes the onset of illumination by a few hours. On the other hand, light affects LI818 mRNA levels independently of active photosynthesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040659
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  • 159
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    Salt-inducible betaine aldehyde dehydrogenase from sugar beet: cDNA cloning and expression (1992)
    Springer
    Plant molecular biology 18 (1992), S. 1-11 
    Details
    ISSN: 1573-5028
    Keywords: betaine aldehyde dehydrogenase ; gene expression ; glycine betaine ; osmotic stress ; salt tolerance ; sugar beet
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Members of the Chenopodiaceae, such as sugar beet and spinach, accumulate glycine betaine in response to salinity or drought stress. The last enzyme in the glycine betaine biosynthetic pathway is betaine aldehyde dehydrogenase (BADH). In sugar beet the activity of BADH was found to increase two- to four-fold in both leaves and roots as the NaCl level in the irrigation solution was raised from 0 to 500 mM. This increase in BADH activity was paralleled by an increase in level of translatable BADH mRNA. Several cDNAs encoding BADH were cloned from a λgt10 libary representing poly(A)+ RNA from salinized leaves of sugar beet plants, by hybridization with a spinach BADH cDNA. Three nearly full-length cDNA clones were confirmed to encode BADH by their nucleotide and deduced amino acid sequence identity to spinach BADH; these clones showed minor nucleotide sequence differences consistent with their being of two different BADH alleles. The clones averaged 1.7 kb and contained an open reading frame predicting a polypeptide of 500 amino acids with 83% identity to spinach BADH. RNA gel blot analysis of total RNA showed that salinization to 500 mM NaCl increased BADH mRNA levels four-fold in leaves and three-fold in the taproot. DNA gel blot analyses indicated the presence of at least two copies of BADH in the haploid sugar beet genome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00018451
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  • 160
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    Developmental and environmental concurrent expression of sunflower dry-seed-stored low-molecular-weight heat-shock protein and Lea mRNAs (1992)
    Springer
    Plant molecular biology 19 (1992), S. 781-792 
    Details
    ISSN: 1573-5028
    Keywords: sunflower ; gene expression ; zygotic embryogenesis ; Lea proteins ; heat-shock proteins ; abscisic acid ; osmotic stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have cloned and sequenced three different cDNAs from sunflower seed-stored mRNA. Sequence similarities and response to heat-shock identified one of the cDNAs as a low-molecular-weight heat-shock protein (lmw-HSP). The other two clones showed significant sequence similarity to the cotton and carrot late-embryogenesis-abundant (Lea) proteins D-113 and Emb-1, respectively. The three cDNAs showed similar expression patterns during zygotic embryo development, as well as in vegetative tissues of 3-day-old seedlings in response to stress. Maximal accumulation of all three mRNAs was detected in dry seeds and during embryo mid-maturation stage, in the absence of exogenous stress. In seedlings, mRNAs accumulated to lower levels in response to osmotic stress and exogenous abscisic acid (ABA) treatments. A differential time course of response to osmotic stress was observed: lmw-HSP mRNA accumulation was induced earlier than that of Lea mRNAs. The coordinate accumulation of Lea and lmw-HSP transcripts during embryo development and in response to stress and ABA suggests the existence of common regulatory elements for Lea and lmw-HSP genes, and supports the notion that HSPs might have alternative functions in the plant cell.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027074
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  • 161
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    Molecular cloning and expression of the gene encoding ADP-glucose pyrophosphorylase from the cyanobacteriumAnabaena sp. strain PCC 7120 (1992)
    Springer
    Plant molecular biology 20 (1992), S. 37-47 
    Details
    ISSN: 1573-5028
    Keywords: ADP-glucose pyrophosphorylase ; Anabaena 7120 ; Escherichia coli ; gene cloning ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previous studies have indicated that ADP-glucose pyrophosphorylase (ADPGlc PPase) from the cyanobacteriumAnabaena sp. strain PCC 7120 is more similar to higher-plant than to enteric bacterial enzymes in antigenicity and allosteric properties. In this paper, we report the isolation of theAnabaena ADPGlc PPase gene and its expression inEscherichia coli. The gene we isolated from a genomic library utilizes GTG as the start codon and codes for a protein of 48347 Da which is in agreement with the molecular mass determined by SDS-PAGE for theAnabaena enzyme. The deduced amino acid sequence is 63, 54, and 33% identical to the rice endosperm small subunit, maize endosperm large subunit, and theE. coli sequences, respectively. Southern analysis indicated that there is only one copy of this gene in theAnabaena genome. The cloned gene encodes an active ADPGlc PPase when expressed in anE. coli mutant strain AC70R1-504 which lacks endogenous activity of the enzyme. The recombinant enzyme is activated and inhibited primarily by 3-phosphoglycerate and Pi, respectively, as is the nativeAnabaena ADPGlc PPase. Immunological and other biochemical studies further confirmed the recombinant enzyme to be theAnabaena enzyme.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029147
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  • 162
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    Nucleotide sequence and temporal regulation of a seed-specificBrassica napus cDNA encoding a stearoyl-acyl carrier protein (ACP) desaturase (1992)
    Springer
    Plant molecular biology 20 (1992), S. 151-155 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; desaturation ; oil synthesis ; embryogenesis ; stearoyl-acyl carrier protein desaturase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The nucleotide sequence is reported for a cDNA containing the entire coding region of a stearoyl-ACP desaturase (EC 1.14.99.6) fromBrassica napus L. cv. Jet neuf. The cDNA was obtained from a library constructed from poly(A)+ RNA purified from embryo tissue. The derived amino acid sequence demonstrates substantial similarity with those from other plant Δ9-desaturases. Comparative RNA-dot blot analyses using the Δ9-desaturase cDNA and a rapessed oleosin cDNA as probes showed that although both these transcripts were seed-specific, they exhibited distinct patterns of temporal regulation. The desaturase message was induced by 25 days after anthesis (DAA), peaking at 45 DAA but decreasing considerably thereafter. In contrast, the oleosin transcript did not increase until 45–50 DAA, reaching a peak much later at about 70 DAA.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00029157
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  • 163
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    Differential accumulation of mRNAs encoding extracellular and intracellular PR proteins in tomato induced by virulent and avirulent races of Cladosporium fulvum (1992)
    Springer
    Plant molecular biology 20 (1992), S. 513-527 
    Details
    ISSN: 1573-5028
    Keywords: β-1,3-glucanase ; gene expression ; pathogenesis-related proteins ; plant-fungus interaction ; protein P14
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tomato leaves infected by the fungal pathogen Cladosporium fulvum contain several types of intracellular and extracellular pathogenesis-related (PR) proteins. Previously, we reported the purification and serological characterization of five extracellular PR proteins: P2, P4, P6, a chitinase and a β-1,3-glucanase [22, 23]. Here we describe the purification of a basic intracellular 33 kDa β-1,3-glucanase and the isolation and characterization of cDNA clones encoding the two extracellular P14 isomers P4 and P6, the extracellular acidic β-1,3-glucanase and a basic 35 kDa β-1,3-glucanase, different from the purified 33 kDa protein. Southern blot analysis demonstrated that tomato PR proteins are not encoded by large gene families, as is the case in tobacco. The number of genes corresponding to each protein was estimated to vary between one and three. A northern blot analysis indicated that the mRNAs for the extracellular PR proteins (P4, P6 and acidic β-1,3-glucanase) accumulate to similar levels in compatible and incompatible tomato-C. fulvum interactions, although the maximum level of expression is reached much faster in the incompatible interaction. On the other hand, the mRNA for the basic 35 kDa β-1,3-glucanase is induced rapidly to high levels in both interactions, but declines in time to background levels only in the incompatible interaction. The relevance of this difference in relation to plant defence is discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040610
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    Expression and sequence analysis of cDNAs induced during the early stages of tuberisation in different organs of the potato plant (Solanum tuberosum L.) (1992)
    Springer
    Plant molecular biology 20 (1992), S. 641-651 
    Details
    ISSN: 1573-5028
    Keywords: S-adenosylmethionine decarboxylase ; differential screening ; gene expression ; Solanum tuberosum ; tuberisation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract cDNA clones of two genes (TUB8 and TUB13) which show a 25–30-fold increase in transcript in the stolon tip during the early stages of tuberisation, have been isolated by differential screening. These genes are also expressed in leaves, stems and roots and the expression pattern in these organs changes on tuberisation. Southern analysis shows homologous sequences in the non-tuberising wild type potato species Solanum brevidens and in Lycopersicon esculentum (tomato). Sequence analysis reveals a high degree of similarity between the TUB13 cDNA, and a human S-adenosylmethionine decarboxylase gene. The predicted TUB8 peptide sequence shows several repeats of alanine, glutamate and proline which suggests a structural role for the encoded protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00046449
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  • 165
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    Structure and expression of a sugarcane gene encoding a housekeeping phosphoenolpyruvate carboxylase (1992)
    Springer
    Plant molecular biology 20 (1992), S. 663-671 
    Details
    ISSN: 1573-5028
    Keywords: PEP carboxylase ; housekeeping gene ; gene expression ; gene structure ; light-mediated activation ; Saccharum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A gene (SCPEPCD1) encoding phosphoenolpyruvate carboxylase (PEPC) was isolated from the C-4 monocot sugarcane (Saccharum hybrid var. H32-8560). SCPEPCD1 is ca. 6800 bp long, with 10 exons. The entire gene sequence from −1561 to 262 bp downstream of the putative poly(A) addition signal is reported. A low-level, essentially constitutive pattern of expression, amino acid sequence similarities to other ‘housekeeping’ PEPC enzymes, and the absence of DNA sequence elements conserved in the upstream region of maize and sorghum C-4-specific PEPC genes indicate that SCPEPCD1 encodes a housekeeping PEPC. Despite this, a motif proposed to act as a phosphorylation site in light-mediated activation of photosynthetic PEPC enzymes [10] is present in the SCPEPCD1 protein; evidence is presented for the presence of this site in other housekeeping PEPC proteins.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00046451
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  • 166
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    Molecular cloning of an alanine aminotransferase from NAD-malic enzyme type C4 plant Panicum miliaceum (1992)
    Springer
    Plant molecular biology 20 (1992), S. 705-713 
    Details
    ISSN: 1573-5028
    Keywords: alanine aminotransferase ; C4 photosynthesis ; gene expression ; nucleotide sequencing ; Panicum miliaceum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have determined the nucleotide sequence of a cDNA encoding AlaAT-2, which is believed to function in the C4-pathway of Panicum miliaceum. An open reading frame (1446 bp) encodes a protein of 482 amino acid residues. The deduced amino acid sequence of AlaAT-2 shows 44.2 and 44.8% homology with the amino acid sequences of AlaATs from rat and human livers, respectively. Northern blot analysis showed that the gene encoding AlaAT-2 in mesophyll and bundle sheath cells was the same and transcribed similarly in the cells. The level of translatable mRNA for AlaAT-2 increased dramatically during greening.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00046455
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  • 167
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    Dynamical behavior of psb gene transcripts in greening wheat seedlings. I. Time course of accumulation of the psbA through psbN gene transcripts during light-induced greening (1992)
    Springer
    Plant molecular biology 20 (1992), S. 695-704 
    Details
    ISSN: 1573-5028
    Keywords: chloroplast ; gene expression ; photosystem 2 ; transcription ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The time course of the accumulation of the transcripts from 13 psb genes encoding a major part of the proteins composing photosystem II during light-induced greening of dark-grown wheat seedlings was examined focusing on early stages of plastid development (0.5 h through 72 h). The 13 genes can be divided into three groups. (1) The psbA gene is transcribed as a single transcript of 1.3 kb in the dark-grown seedlings, but its level increases 5- to 7-fold in response to light due to selective increase in RNA stability as well as in transcription activity. (2) The psbE-F-L-J operon, psbM and psbN genes are transcribed as a single transcript of 1.1 kb, two transcripts of 0.5 and 0.7 kb and a single transcript of 0.3 kb, respectively, in the dark-grown seedlings. The levels of accumulation of every transcript remain unchanged or rather decrease during plastid development under illumination. (3) The psbK-I-D-C gene cluster and psbB-H operon exhibit fairly complicated northern hybridization patterns during the greening process. When a psbC or psbD gene probe was used for northern hybridization, five transcripts differing in length were detected in the etioplasts from 5-day old dark-grown seedlings. After 2 h illumination, two new transcripts of different length appeared. Light induction of new transcripts was also observed in the psbB-H operon.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00046454
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    Effect of salt stress on plant gene expression: A review (1992)
    Springer
    Plant and soil 146 (1992), S. 145-151 
    Details
    ISSN: 1573-5036
    Keywords: barley ; gene expression ; ice plant ; rice ; salt stress ; tobacco ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract Soil salinity is an important agricultural problem, particularly since the majority of crop plants have low salt tolerance. The identification of genes whose expression enables plants to adapt to or tolerate salt stress is essential for breeding programs, but little is known about the genetic mechanisms for salt tolerance. Recent research demonstrates that salt stress modulates the levels of a number of gene products. Although the detection of gene products that respons specifically to salt stress is a significant finding, they must be identified, functions assigned, and their relation to salt tolerance determined. This article focuses on a few of the salt-responsive proteins and mRNAs that have been discovered and the methods employed to identify and characterize them.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00012007
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  • 169
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    Accumulation of wound-inducible ACC synthase transcript in tomato fruit is inhibited by salicylic acid and polyamines (1992)
    Springer
    Plant molecular biology 18 (1992), S. 477-487 
    Details
    ISSN: 1573-5028
    Keywords: Tomato ; gene expression ; wounding ; ethylene ; glycine-rich protein ; rRNA ; polyamines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Regulation of wound-inducible 1-aminocyclopropane-1-carboxylic acid (ACC) synthase expression was studied in tomato fruit (Lycopersicon esculentum cv. Pik-Red). A 70 base oligonucleotide probe homologous to published ACC synthase cDNA sequences was successfully used to identify and analyze regulation of a wound-inducible transcript. The 1.8 kb ACC synthase transcript increased upon wounding the fruit as well as during fruit ripening. Salicylic acid, an inhibitor of wound-responsive genes in tomato, inhibited the wound-induced accumulation of the ACC synthase transcript. Further, polyamines (putrescine, spermidine and spermine) that have anti-senescence properties and have been shown to inhibit the development of ACC synthase activity, inhibited the accumulation of the wound-inducible ACC synthase transcript. The inhibition by spermine was greater than that caused by putrescine or spermidine. The transcript level of a wound-repressible glycine-rich protein gene and that of the constitutively expressed rRNA were not affected as markedly by either salicylic acid or polyamines. These data suggest that salicylic acid and polyamines may specifically regulate ethylene biosynthesis at the level of ACC synthase transcript accumulation.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040664
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  • 170
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    Effect of two consensus sequences preceding the translation initiator codon on gene expression in plant protoplasts (1992)
    Springer
    Plant molecular biology 18 (1992), S. 815-818 
    Details
    ISSN: 1573-5028
    Keywords: expression cassette ; gene expression ; protoplasts ; translation initiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Expression cassettes containing a duplicated cauliflower mosaic virus (CaMV) 35S promoter fused to a polylinker preceded by the CCACCATGG and AACAATGG sequences were constructed. These two sequences correspond to the consensus sequences around the translation start codons in vertebrates and plants respectively. Translational fusions were made with the β-glucuronidase-coding sequence and transient expression was recorded in tobacco mesophyll protoplasts. Approximately three times more GUS activity was found in protoplasts incubated with the constructs harbouring translational fusions as compared to a control harbouring a transcriptional fusion. No significant difference was observed between GUS activities obtained with the two consensus sequences.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00020027
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  • 171
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    Structure of a rice β-glucanase gene regulated by ethylene, cytokinin, wounding, salicylic acid and fungal elicitors (1992)
    Springer
    Plant molecular biology 18 (1992), S. 33-45 
    Details
    ISSN: 1573-5028
    Keywords: glucanase ; gene expression ; pathogenesis response ; stress response ; plant growth regulators ; gene family
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A rice β-glucanase gene was sequenced and its expression analyzed at the level of mRNA accumulation. This gene (Gns1) is expressed at relatively low levels in germinating seeds, shoots, leaves, panicles and callus, but it is expressed at higher levels in roots. Expression in the roots appears to be constitutive. Shoots expressGns1 at much higher levels when treated with ethylene, cytokinin, salicylic acid, and fungal elicitors derived from the pathogenSclerotium oryzae or from the non-pathogenSaccharomyces cereviseae. Shoots also expressGns1 at higher levels in response to wounding. Expression in the shoots is not significantly affected by auxin, gibberellic acid or abscisic acid. The β-glucanase shows 82% amino acid similarity to the barley 1,3;1,4-β-D-glucanases, and from hybridization studies it is the β-glucanase gene in the rice genome closest to the barley 1,3;1,4-β-glucanase EI gene. The mature peptide has a calculated molecular mass of 32 kDa. The gene has a large 3145 bp intron in the codon for the 25th amino acid of the signal peptide. The gene exhibits a very strong codon bias of 99% G+C in the third position of the codon in the mature peptide coding region, but only 61% G+C in the signal peptide region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00018454
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  • 172
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    Characterization and expression of U1snRNA genes from potato (1992)
    Springer
    Plant molecular biology 19 (1992), S. 959-971 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; gene variants ; pre-mRNA splicing ; pseudogenes ; U1 small nuclear RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract U1 small nuclear RNAs (U1snRNAs) occur in the nucleus of plants and animals where, complexed with several proteins in the form of U1 small nuclear ribonucleoprotein particles (U1snRNPs), they play an important role in precursor messenger RNA (pre-mRNA) splicing. Ten potato U1snRNA genes have been isolated on two genomic clones illustrating the clustering of this multigene family on the potato genome. Based on both the sequence of their coding regions and upstream regulatory elements, seven of the genes are potentially functional. The other three genes were pseudogenes with defective promoter or coding region sequences. Analysis of expression of individual cloned U1snRNA genes in transfected tobacco protoplasts was impossible due to the similarity of U1snRNA sequences in tobacco. However, by marking the coding regions with oligonucleotides or constructing chimaeric genes consisting of a potato U1snRNA promoter region and maize U5snRNA coding region, three of the U1 promoter regions were shown to be transcriptionally active.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040528
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  • 173
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    Characterization of the U3 and U6 snRNA genes from wheat: U3 snRNA genes in monocot plants are transcribed by RNa polymerase III (1992)
    Springer
    Plant molecular biology 19 (1992), S. 973-983 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; promoter specificity ; snRNA gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have demonstrated recently that the genes encoding the U3 small nuclear RNA (snRNA) in dicot plants are transcribed by RNA polymerase III (pol III), and not RNA polymerase II (pol II) as in all other organisms studied to date. The U3 gene was the first example of a gene transcribed by different polymerases in different organisms. Based on phylogenetic arguments we proposed that a polymerase specificity change of the U3 snRNA gene promoter occurred during plant evolution. To map such an event we are examining the U3 gene polymerase specificity in other plant species. We report here the characterization of a U3 gene from wheat, a monocot plant. This gene contains the conserved promoter elements, USE and TATA, in a pol III-specific spacing seen also in a wheat U6 snRNA gene characterized in this report. Both the U3 and the U6 genes possess typical pol III termination signals but lack the cis element, responsible for 3′-end formation, found in all plant pol II-specific snRNA genes. In addition, expression of the U3 gene in transfected maize protoplasts is less sensitive to α-amanitin than a pol II-transcribed U2 gene. Based on these data we conclude that the wheat U3 gene is transcribed by pol III. This observation suggests that the postulated RNA polymerase specificity switch of the U3 gene took place prior to the divergence of angiosperm plants into monocots and dicots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040529
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  • 174
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    Molecular analysis of a cruciferin storage protein gene family of Brassica napus (1992)
    Springer
    Plant molecular biology 19 (1992), S. 1049-1055 
    Details
    ISSN: 1573-5028
    Keywords: Brassica napus ; rapeseed ; gene expression ; nucleotide sequence ; storage proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated a five-member gene subfamily which encodes cruciferin, a legumin-like 12S storage protein of Brassica napus L., and have analyzed the structure and expression of the family members in developing embryos. Sequence analysis has shown that the coding regions of all five genes are highly similar, with the two most divergent members of the family retaining 89% sequence identity. The analysis of this cruciferin gene family's expression indicates that the developmental pattern of expression of each gene is similar, and the steady-state mRNA levels of each gene are approximately equivalent to each other at all developmental stages.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040536
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  • 175
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    Differential gene expression during germination and after the induction of adventitious bud formation in Norway spruce embryos (1992)
    Springer
    Plant molecular biology 18 (1992), S. 713-724 
    Details
    ISSN: 1573-5028
    Keywords: adventitious buds ; cDNA cloning ; cytokinin ; gene expression ; germination ; Norway spruce
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A pulse treatment of embryos of Norway spruce with cytokinin suppresses germinative development and induces the coordinate formation of adventitious buds from subepidermal cell layers. To analyse the patterns of gene expression associated with germination and the alterations induced by the bud induction treatment, we have isolated cDNA clones corresponding to genes that are differentially expressed in cytokinin-treated and untreatedin vitro germinating embryos. One category of 14 clones hybridized to transcripts that were abundant specifically during germination. The expression of 8 of these genes was reduced by the bud induction treatment. Four clones, including one identified as a histone H2A gene, recognized transcripts that showed an increased abundance in bud-induced versusin vitro germinating embryos. A second category of 13 clones hybridized to transcripts that increased in abundance during post-germinative development of the seedling. Among these a subset of 8 clones, including an α-tubulin clone, corresponds to genes suppressed by the bud induction treatment, whereas 5 clones, including a gene with sequence similarity to polyubiquitin, were unaffected by the treatment. One clone hybridized to a message abundant in the seed, during early germination as well as in the vegetative bud, and showed 60% partial sequence identity to a barley (1→3)-β-glucanase gene. Genes expressed exclusively in bud-induced orin vitro germinating embryos were not found. The results show that a major difference in gene expression between treated and untreated embryos is related to the shift from extensive cell proliferation to elongation and differentiation that occurs at the transition from germination to post-germinative development, and which is suppressed in the bud-induced embryos.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00020013
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  • 176
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    A probable lipid transfer protein gene is induced by NaCl in stems of tomato plants (1992)
    Springer
    Plant molecular biology 18 (1992), S. 749-757 
    Details
    ISSN: 1573-5028
    Keywords: salt stress ; stem-specific expression ; lipid transfer protein ; cDNA sequence ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A full-length tomato cDNA clone, TSW12, which is developmentally and environmentally regulated, has been isolated and characterized. TSW12 mRNA is accumulated during tomato seed germination and its level increases after NaCl treatment or heat shock. In mature plants, TSW12 mRNA is only detected upon treatment with NaCl, mannitol or ABA and its expression mainly occurs in stems. The nucleotide sequence of TSW12 includes an open reading frame coding for a basic protein of 114 amino acids; the first 23 amino acids exhibit the sequence characteristic of a signal peptide. The high similarity between the TSW12-deduced amino acid sequence and reported lipid transfer proteins suggests that TSW12 encodes a lipid transfer protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00020016
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  • 177
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    Cytoplasmic ribosomal protein S15a from Brassica napus: Molecular cloning and developmental expression in mitotically active tissues (1992)
    Springer
    Plant molecular biology 18 (1992), S. 909-919 
    Details
    ISSN: 1573-5028
    Keywords: ribosomal protein S15a ; cDNA clones ; rapessed ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated two cDNA clones which appear to encode the 40S ribosomal subunit protein S15a from Brassica napus (oilseed rape). The open reading frame in both clones contains 390 bases, encoding a deduced polypeptide sequence of 130 amino acids (100% homology between clones) with 76% sequence identity to the N-terminal 37 amino acids of the rat ribosomal protein S15a and 80% identity to the S24 polypeptide of yeast. Both the yeast and rapeseed proteins have a net positive charge of +9 and the rapeseed S15a protein has a molecular mass of 14778 Da compared to 14762 Da for the yeast protein. The rapeseed ribosomal protein S15a is encoded by a small multi-gene family with at least two actively transcribed members. A single transcript of ca. 1.0 kb, corresponding to ribosomal protein S15a, is abundant in actively dividing tissues such as apical meristem, flower buds and young leaves and less abundant in mature stem and fully expanded leaves.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019205
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  • 178
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    Expression of 17 rye (Secale cereale L.) traits in a range of durum wheat (Triticum durum Desf.) and bread wheat (T. aestivum L.) genetic backgrounds (1992)
    Springer
    Euphytica 60 (1992), S. 37-44 
    Details
    ISSN: 1573-5060
    Keywords: Triticum aestivum ; Triticum durum ; bread wheat ; durum wheat Secale cereale ; rye ; gene expression ; alien introduction
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Expression of 17 rye traits in 24 bread wheat x rye and 8 durum wheat x rye crosses was studied, using a self-compatible, homozygous, dwarf rye. Rye showed epistasis for hairiness on the peduncle in all the crosses of Triticum aestivum and T. durum wheats with rye. Dark greenness of leaves of rye was expressed in all the durum wheat x rye and in some of the bread wheat x rye crosses. Similarly, absence of auricle pubescence, a rye trait, was expressed in most of the durum wheat x rye crosses but not in the bread wheat x rye crosses, indicating the presence of inhibitors for these traits frequently on the D genome and rarely on the A and/or B genome of wheat. Most of the wide hybrids resembled rye fully or partially for intense waxy bloom on the leaf-sheath and for the absence of basal underdeveloped spikelets. Similarly, most of the amphihaploids resembled rye for the anthocyanin in the coleoptile, stem and node. The presence of some inhibitors on A and/or B genome of wheat was indicated in some of the wheat genotypes for the expression of rye traits viz. intense waxy bloom, anthocyanin in node and absence of basal underdeveloped spikelets. Enhancement in the level of expression of the intensity and length of bristles on the mid-rib of the glume of the hybrids might be due to wheat-rye interaction. Less number of florets/spikelet as in rye showed variable expression in different wheat backgrounds. Some other rye traits like absence of auricles, terminal spikelet and glume-awn were not expressed in the wheat background. The expression of some of the rye genes might have been influenced by their interaction with Triticum cytoplasm and/or the environment.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00022256
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  • 179
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    Isolation and characterization of a cDNA that encodes ECP31, an embryogenic-cell protein from carrot (1992)
    Springer
    Plant molecular biology 19 (1992), S. 239-249 
    Details
    ISSN: 1573-5028
    Keywords: ABA ; Daucus carota ; ECP31 ; gene expression ; LEA clone ; somatic embryogenesis
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A full-length cDNA for ECP31, an embryogenic cell protein from carrot (Daucus carota L.) with a M r of 31000 (Kiyosue T, Satoh S, Kamada H, Harada H (1991) Plant Physiol 95: 1077–1083), was isolated from a cDNA library prepared from embryogenic cells using PCR-amplified DNA as a probe. The genomic Southern blot analysis revealed that there are two or three genes for ECP31 in the carrot genome. The transcripts of ECP31 accumulated in the peripheral regions of clusters of embryogenic cells and disappeared in the course of somatic embryogenesis that was induced by transfer of the embryogenic cells to auxin-free media. The cDNA encodes a polypeptide of 256 amino acids, and the calculated molecular weight of this polypeptide is 26 111. The deduced amino acid sequence shows a high degree (62.2%) of similarity to that of a protein that is abundant during late embryogenesis of cotton (LEA D34; Baker JC, Steele C, Dure III (1988) Plant Mol Biol 11: 227–291). The mRNAs for ECP31 started to accumulate in zygotic embryos at a late stage of embryogenesis but were undetectable in mature embryos within 24 h after imbibition of seeds. In dry fruits (seeds), the transcripts were detected only in zygotic embryos by in situ hybridization. The level of ECP31 transcripts increased after treatment with abscisic acid (ABA) in torpedo-shaped somatic embryos but not in seven-day-old seedlings. These results suggest that both embryo-specific factor(s) and ABA are involved in the expression of the gene for ECP31.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027345
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  • 180
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    Nucleotide sequence of a tobacco cDNA encoding plastidic glutamine synthetase and light inducibility, organ specificity and diurnal rhythmicity in the expression of the corresponding genes of tobacco and tomato (1992)
    Springer
    Plant molecular biology 19 (1992), S. 367-379 
    Details
    ISSN: 1573-5028
    Keywords: cDNA sequence ; gene expression ; glutamine synthetase ; phytochrome ; Solanaceae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A full-length cDNA encoding glutamine synthetase (GS) was cloned from a λgt10 library of tobacco leaf RNA, and the nucleotide sequence was determined. An open reading frame accounting for a primary translation product consisting of 432 amino acids has been localized on the cDNA. The calculated molecular mass of the encoded protein is 47.2 kDa. The predicted amino acid sequence of this precursor shows higher homology to GS-2 protein sequences from other species than to a leaf GS-1 polypeptide sequence, indicating that the cDNA isolated encodes the chloroplastic isoform (GS-2) of tobacco GS. The presence of C-and N-terminal extensions which are characteristic of GS-2 proteins supports this conclusion. Genomic Southern blot analysis indicated that GS-2 is encoded by a single gene in the diploid genomes of both tomato and Nicotiana sylvestris, while two GS-2 genes are very likely present in the amphidiploid tobacco genome. Western blot analysis indicated that in etiolated and in green tomato cotyledons GS-2 subunits are represented by polypeptides of similar size, while in green tomato leaves an additional GS-2 polypeptide of higher apparent molecular weight is detectable. In contrast, tobacco GS-2 is composed of subunits of identical size in all organs examined. GS-2 transcripts and GS-2 proteins could be detected at high levels in the leaves of both tobacco or tomato. Lower amounts of GS-2 mRNA were detected in stems, corolla, and roots of tomato, but not in non-green organs of tobacco. The GS-2 transcript abundance exhibited a diurnal fluctuation in tomato leaves but not in tobacco leaves. White or red light stimulated the accumulation of GS-2 transcripts and GS-2 protein in etiolated tomato cotyledons. Far-red light cancelled this stimulation. The red light response of the GS-2 gene was reduced in etiolated seedlings of the phytochrome-deficient aurea mutant of tomato. These results indicate a phytochrome-mediated light stimulation of GS-2 gene expression during greening in tomato.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023384
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  • 181
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    Structure of the pine (Pinus thunbergii) chlorophyll a/b-binding protein gene expressed in the absence of light (1992)
    Springer
    Plant molecular biology 19 (1992), S. 405-410 
    Details
    ISSN: 1573-5028
    Keywords: cab gene ; chlorophyll a/b-binding protein ; gymnosperm ; gene expression ; pine (Pinus thunbergii)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A gene for chlorophyll a/b-binding protein (cab) of pine (Pinus thunbergii) was isolated and sequenced. The gene (cab-6) contains an intron at a position equivalent to the type II cab genes of angiosperms. Transcript mapping analyses show that the amount of the mRNA in the dark is about half of that in the light. The cab-6 gene is expressed in dark-grown seedlings at a very high level, differing from angiosperm cab genes which are induced by light. The cab-6 gene typifies the coniferous plant cab genes in light-independent gene expression.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00023388
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  • 182
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    The heat shock response of pollen and other tissues of maize (1992)
    Springer
    Plant molecular biology 19 (1992), S. 623-630 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; heat shock ; intron ; maize ; pollen ; RNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract While a heat shock treatment of 40 °C or 45 °C induced the vegetative tissues of maize to produce the typical heat shock proteins (HSPs), germinating maize pollen exposed to the same temperatures did not synthesize these characteristic HSPs. Comparison of RNA accumulation in shoot and tassel tissue showed that mRNAs for HSP70 and HSP18 increased several-fold, reaching high levels within 1 or 2 hours. At the higher temperature of 45 °C these vegetative tissues were blocked in removal of an intron from the HSP70 mRNA precursor, which accumulated to a high level in tassel tissue. In germinating pollen exposed to heat shock, mRNAs for these HSPs were induced but accumulated only to low levels. The stressed pollen maintained high levels of RNA for α-tubulin, a representative normal transcript. It is likely that the defective heat shock response of maize pollen is due to inefficient induction of heat shock gene transcription.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00026788
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  • 183
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    Pea dehydrins: identification, characterisation and expression (1992)
    Springer
    Plant molecular biology 19 (1992), S. 1031-1044 
    Details
    ISSN: 1573-5028
    Keywords: ABA ; dehydrin ; gene expression ; pea (Pisum sativum L.) ; seed development ; water stress
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An antiserum raised against dehydrin from maize (Zea mays) recognised several polypeptides in extracts of pea (Pisum sativum) cotyledons. A cDNA expression library was prepared from mRNA of developing cotyledons, screened with the antiserum and positive clones were purified and characterised. The nucleotide sequence of one such clone, pPsB12, contained an open reading frame which would encode a polypeptide with regions of significant amino acid sequence similarity to dehydrins from other plant species. The deduced amino acid sequence of the pea dehydrin encoded by B12 is 197 amino acids in length, has a high glycine content (25.9%), lacks tryptophan and is highly hydrophilic. The polypeptide has an estimated molecular mass of 20.4 kDa and pI=6.4. An in vitro synthesised product from the clone comigrates with one of the in vivo proteins recognised by the antiserum. A comparison of the pea dehydrin sequence with sequences from other species revealed conserved amino acid regions: an N-terminal DEYGNP and a lysine-rich block (KIKEKLPG), both of which are present in two copies. Unexpectedly, pea dehydrin lacks a stretch of serine residues which is conserved in other dehydrins. B12 mRNA and dehydrin proteins accumulated in dehydration-stressed seedlings, associated with elevated levels of endogenous abscisic acid (ABA). Applied ABA induced expression of dehydrins in unstressed seedlings. Dehydrin expression was rapidly reversed when seedlings were removed from the stress or from treatment with ABA and placed in water. During pea cotyledon development, dehydrin mRNA and proteins accumulated in mid to late embryogenesis. Dehydrin proteins were some of the most actively synthesised at about the time of maximum fresh weight and represent about 2% of protein in mature cotyledons.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040534
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    Temporal and spatial regulation of a novel gene in barley embryos (1992)
    Springer
    Plant molecular biology 20 (1992), S. 255-266 
    Details
    ISSN: 1573-5028
    Keywords: barley (Hordeum vulgare) ; zygotic embryogenesis ; plant development ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The temporal and spatial pattern of expression of a novel barley gene is described. The gene has been identified through the differential screening of a cDNA library constructed to poly(A)+ RNA of zygotic embryos. Transcripts corresponding to the cDNA, pZE40, become abundant in the non-axial tissues of the developing embryo within 8–10 days after anthesis, when steady-state levels are high in the scutellum, coleoptile and coleorhiza, with the exception of the scutellar epithelium. This expression pattern is maintained throughout maturation of the embryo until levels eventually decline as the grain desiccates. On germination, there is a transient re-appearance of mRNA to pZE40, with accumulation specifically restricted to the scutellum of the seedling. In situ hybridization has enabled the detection of transcripts elsewhere in the barley plant, in highly localized groups of cells. The timing and cell specificity of expression suggests the gene product is involved in the synthesis and/or transport of metabolites.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014493
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    Isolation and characterization of a tobacco gene with homology to pectate lyase which is specifically expressed during microsporogenesis (1992)
    Springer
    Plant molecular biology 20 (1992), S. 493-502 
    Details
    ISSN: 1573-5028
    Keywords: gene expression ; microsporogenesis ; Nicotiana tabacum ; pectate lyase ; pollen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A genomic clone has been isolated which contains an open reading frame of 1191 bp interrupted by two small introns. The ORF has been sequenced and the transcriptional start determined. The predicted amino acid sequence shows homology to the deduced amino acid sequences of two pollen-specific pectate lyase genes identified in tomato. The genomic clone was isolated using a partial cDNA clone, TP10, which had been isolated from a Nicotiana tabacum pollen cDNA library by means of differential screening. TP10 has been fully sequenced and contains an open reading frame of 792 bp which shows 96% homology to the ORF in the genomic clone. The transcript corresponding to TP10 is maximally expressed late in pollen development, and has not been detected in vegetative tissues.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040608
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  • 186
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    cDNA cloning of a tetraubiquitin gene, and expression of ubiquitin-containing transcripts, in aleurone layers of Avena fatua (1992)
    Springer
    Plant molecular biology 20 (1992), S. 753-758 
    Details
    ISSN: 1573-5028
    Keywords: aleurone ; Avena fatua ; cDNA nucleotide sequence ; gene expression ; gibberellin ; polyubiquitin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A λgt11 cDNA library, constructed from poly(A)+ mRNA isolated from Avena fatua aleurone layers incubated with 1 μM gibberellin A1 (GA1) for 4 days, was screened with an anti-idiotypic antiserum raised against the GA-specific monoclonal antibody MAC 182. One positive clone was isolated, sequenced and shown to encode a tetraubiquitin based on the deduced amino acid sequence. This polyubiquitin cDNA exhibited a high degree of homology to a cloned wheat hexaubiquitin in its 3′-non-coding region. Analysis of total RNA isolated from A. fatua aleurone layers, treated without or with a range of concentrations of GA1 from 10-11 to 10-6 M, by northern blotting using the cDNA probe revealed 8 different ubiquitin-containing transcript classes all of which are constitutively expressed in aleurone and are regulated by GA1.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00046461
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  • 187
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    Ubiquitin genes are differentially regulated in protoplast-derived cultures of Nicotiana sylvestris and in response to various stresses (1992)
    Springer
    Plant molecular biology 20 (1992), S. 897-910 
    Details
    ISSN: 1573-5028
    Keywords: Cell division ; gene expression ; Nicotiana sylvestris ; protoplast ; stress ; ubiquitin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Four ubiquitin mRNA size classes were found to be differentially regulated in mesophyll protoplast-derived cultures of Nicotiana sylvestris. Three mRNA families of 1.9, 1.6 and 1.35 kb were expressed as soon as protoplasts were isolated. The 1.9 and 1.6 kb size classes were transiently expressed during the first hours of culture, whereas the level of expression of the 1.35 kb size class was maintained as long as cells kept dividing. A 0.7 kb mRNA size class started to be expressed just before the first divisions were observed. cDNAs corresponding to each of these families were isolated from a 6-h-old protoplast cDNA library and characterized. The 1.9, 1.6 and 1.35 kb mRNAs thus encode 7- or more, 6- and 5- mers, respectively, of ubiquitin whereas the 0.7 kb mRNAs encode a monomer of ubiquitin fused to a carboxyl extension protein of 52 amino acids. The expression of ubiquitin genes was studied, using probes specific for each of these transcript families, during protoplast culture and, for comparison, after various stresses including heat shock, HgCl2 treatment, a viral infection giving rise to a hypersensitive reaction, and an Agrobacterium tumefaciens infection which resulted in tumour formation. The 1.9 and 1.6 kb mRNA size classes were found to be stress-regulated, the 0.7 kb mRNA size class developmentally regulated and the 1.35 kb size class both stress- and developmentally regulated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027161
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  • 188
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    Translation controls the expression level of a chimaeric reporter gene (1992)
    Springer
    Plant molecular biology 20 (1992), S. 921-938 
    Details
    ISSN: 1573-5028
    Keywords: β-D-glucuronidase ; mannopine synthase promoter ; Agrobacterium ; gene expression ; initiation of translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transcriptional and translational fusions between the reading frame of the β-D-glucuronidase gene (gusA) and the 2′ as well as the 1′ promoter of mannopine synthase (mas), a TR locus of Agrobacterium tumefaciens, were made. The expression of these constructs was studied in the transgenic F1 offspring of independent tobacco transformants at the protein level by assaying for GUS activity and western blot analysis of the GUS protein and at the steady-state mRNA level. In leaves, stems and roots no correlation was found between steady-state levels of GUS mRNA and enzyme activity. In older tissues significantly higher GUS activities were found. This is explained by the stable character of the GUS protein together with an accumulation of protein upon ageing. Three to ten times higher GUS activities were found for in vitro grown plants than for greenhouse-grown plants of the same offspring, despite similar levels of GUS mRNA. Roots from in vitro grown plants display three to ten times higher GUS activities than stems and leaves. In transgenic plants grown in vitro, containing a translational fusion with two AUGs in phase, the initiation of translation in leaf material occurred at both AUGs. Initiation of translation at the first AUG, however, was ten times more frequent. In contrast, initiation in roots from in vitro grown plants occurred exclusively at the second AUG.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00027163
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  • 189
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    Electronic Resource
    A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome (1992)
    Springer
    Plant molecular biology 20 (1992), S. 1203-1207 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium ; binary vector ; CaMV 35S ; gene expression ; β-glucuronidase ; Nicotiana plumbaginifolia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A versatile gene expression cartridge and binary vector system was constructed for use in Agrobacterium-mediated plant transformation. The expression cartridge of the primary cloning vector, pART7, comprises of cauliflower mosaic virus Cabb B-JI isolate 35S promoter, a multiple cloning site and the transcriptional termination region of the octopine synthase gene. The entire cartridge can be removed from pART7 as a Not I fragment and introduced directly into the binary vector, pART27, recombinants being selected by blue/white screening for β-galactosidase. pART27 carries the RK2 minimal replicon for maintenance in Agrobacterium, the ColE1 origin of replication for high-copy maintenance in Escherichia coli and the Tn7 spectinomycin/streptomycin resistance gene as a bacterial selectable marker. The organisational structure of the T-DNA of pART27 has been constructed taking into account the right to left border, 5′ to 3′ model of T-DNA transfer. The T-DNA carries the chimaeric kanamycin resistance gene (nopaline synthase promoter-neomycin phosphotransferase-nopaline synthase terminator) distal to the right border relative to the lacZ′ region. Utilisation of these vectors in Agrobacterium-mediated transformation of tobacco demonstrated efficient T-DNA transfer to the plant genome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028910
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  • 190
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    Ascorbic acid upregulates myelin gene expression in C6 glioma cells (1992)
    Springer
    Metabolic brain disease 7 (1992), S. 157-164 
    Details
    ISSN: 1573-7365
    Keywords: ascorbic acid ; proteolipid protein (PLP) ; myelin associated glycoprotein (MAG) ; gene expression ; C6 glioma cells
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effect of ascorbic acid (AA) on rat glioma C6 cells was studied. At physiological AA concentrations of 0.1 and 1 mM, no morphological and no proliferative alterations in the C6 cultures were detectable. Although the total RNA content per cell was not affected by the AA-treatment, AA upregulated the expression of myelin-specific genes, i.e. proteolipid protein (PLP) and myelin-associated glycoprotein (MAG) genes as assessed by northern blot analysis. The steady-state level of the specific mRNAs increased transiently in the AA-treated cells. Three days after AA administration the message level reached a maximum of 10-and 2-fold over control for the PLP and MAG genes, respectively. The upregulation of the genes was directly related to AA concentration. The present data indicate a possible involvement of AA in the regulation of myelin gene activity in the CNS.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01000161
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  • 191
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    Increased calmodulin affects cell morphology and mRNA levels of cytoskeletal protein genes (1992)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 21 (1992), S. 45-57 
    Details
    ISSN: 0886-1544
    Keywords: cell shape ; gene expression ; pleiotropic effects ; cell cycle ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have previously described stable mouse C127 cell lines in which a CaM mini-gene has been expressed in a bovine papilloma virus-based expression vector (Rasmussen and Means: EMBO J. 6:3961-3968. 1987). Elevation of CaM to levels five-fold higher than in control cells caused an acceleration in cell cycle progression by reducing the length of the G1 period. When these cell lines were originally isolated it was observed that cells in which CaM levels were increased had a flattened morphology. In this study we have examined the localization of actin, vimentin, and tubulin in these cells as compared to the BPV-transformed control cell line in order to determine if changes in shape were accompanied by differences in the cytoskeletal organization. Cell-cycle-dependent changes in the levels of mRNAs for histone H4, glyceraldehyde-3-phosphate dehydrogenase, β-actin, vimentin, and β-tubulin have also been examined. Our results indicate that increased CaM causes differences in the organization of microfilaments, intermediate filaments, and microtubules and that these changes are accompanied by selective differences in the cell-cycle-dependent expression of some mRNAs. Elevated CaM was also correlated with a reduced stability of β-tubulin mRNA. These studies indicate that CaM has pleiotropic effects on cell function and suggest that stable cell lines with altered CaM levels may provide a useful model system for understanding the moiecular basis of CaM-dependent regulation of cellular processes.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970210106
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  • 192
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    Comparative analysis of the β transducin family with identification of several new members including PWP1, a nonessential gene of Saccharomyces cerevisiae that is divergently transcribed from NMT1 (1992)
    New York, NY : Wiley-Blackwell
    Proteins: Structure, Function, and Genetics 13 (1992), S. 41-56 
    Details
    ISSN: 0887-3585
    Keywords: fungal genes ; molecular cloning ; molecular sequence data ; gene expression ; database searching ; sequence alignment ; repetitive sequences ; signal transduction ; protein N-myristoylation ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: While investigating the expression of the Saccharomyces cerevisiae myristoyl-CoA:protein N-myristoyltransferase gene (NMT: E.C. 2.3.1.97) by Northern blot analysis, we observed another RNA transcript whose expression resembled that of NMT1 during meiosis and was derived from a gene located 〈1 kb immediately upstream of NMT1. This new gene, designated PWP1 (for periodic tryptophan protein), is divergently transcribed from NMT1 and encodes a 576-residue protein. Null mutants of PWP1 are viable, but their-growth is severely retarded and steady-state levels of several cellular proteins (including at least two proteins that label with exogenous [3H]myristic acid) are drastically reduced. New methods for database searching and assessing the statistical significance of sequence similarities identic PWP1 as a member of the β-transducin protein superfamily. Two other previously unrecognized β-transducin-like proteins (S. cerevisiae MAKI1 and D. discoideum AAC3) were also identified, and an unexpectedly high degree of sequence homology was found between a Chlamydomonas β-like polypeptide and the C12.3 gene of chickens. A systematic and quantitative comparative analysis resulted in classifying all β-transducin-like sequences into II nonorthologous families. Based on specific sequence attributes, however, not all β-transducin-like sequences are expected to be functionally similar, and quantitative criteria for inferring functional analogies are discussed. Possible roles of repetitive tryptophan residues in proteins are also considered. Published 1992 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/prot.340130105
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  • 193
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    An adjustable expression system for controlling growth rate, plasmid maintenance, and culture dynamics (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Biotechnology and Bioengineering 40 (1992), S. 1027-1038 
    Details
    ISSN: 0006-3592
    Keywords: continuous fermentation ; pili ; plasmid segregation ; gene expression ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Recombinant bacterial cells express various levels of model product proteins if the genes of interest are regulated by controllable promoters. The level of gene expression influences the growth-rate differential between plasmid-bearing and plasmid-free cells, and thereby affects the culture dynamics of a plasmid-containing cell population. An expression system has been designed in which host Escherichia coli cells contain the pil operon controlled by a tac promoter; these cells are transformed with plasmids that contain the repressor gene, lacl, for the tac promoter, in combination with an expression system for a model protein, chloramphenicol acetyl tranferase (CAT). Experimental and theoretical results show that plasmid-bearing cells can be maintained as dominant in continuous cultures without selective pressure when 12% or less of the cells' total protein is the model product protein, CAT. This is because the segment cells produce pili greatly in excess of normal wild-type levels, and thus have more of a metabolic burden than do the plasmid-bearng cells that overproduce CAT. However, when the level of the plasmid-directed CAT expression is increased above 12% of the cells' total protein, the growth rate of the plasmid-bearing cells decreases to a value lower than that of the segregant cells. Therefore, plasmid-containing cells lose their selective advantage at this expression level, and cannot be maintained as the dominant cell type in a continuous culture unless antibiotic or other positive selection methods are used. By controlling the growth rate differential of this bacterial host/plasmid system, a variety of interesting competitive culture dynamics is investigated. All experimental measurements for continuous cultures are in very good agreement with theory using kinetic parameters determined from independent batch experiments. © 1992 John Wiley & Sons, Inc.
    Additional Material: 14 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bit.260400906
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  • 194
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    Stable production of an analog of human tissue plasminogen activator from culturedDrosophila cells (1992)
    Springer
    Cytotechnology 10 (1992), S. 157-167 
    Details
    ISSN: 1573-0778
    Keywords: Drosophila cell culture ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract We have studied the expression of an analog of human tissue plasminogen activator, FK2P, inDrosophila Schneider 2 cells. A number of promoters were tested, including theDrosophila metallothionein promoter (MTd), baculovirus immediate early promoter (IE),Drosophila copia promoter, mouse metallothionein promoter, cytomegalovirus immediate early promoter with or without intron, SV40 immediate early promoter, and human elongation factor 1α promoter. Two of these promoters drove significant expression of FK2P. The MTd promoter is tightly regulated and upon induction with copper or cadmium expression of FK2P increases as much as 180-fold, accumulating in the culture medium to about 7 μg FK2P/106 cells/day as determined by ELISA. The IE promoter can direct the constitutive expression to yield about 0.4 μg FK2P/106 cells/day. The production of FK2P in these cell lines remains at about the same level after repeated passages, even in the absence of selective pressure. The FK2P accumulated in the culture medium is fully active in an assay using a chromogenic substrate for serine proteases. Western immunoblot analysis shows that the product remains predominately as single-chain molecules in serum-free medium, while in serum-containing medium two-chain material occurs as expected due to the presence of plasmin in serum. Judged from the size in Western immunoblots, the FK2P produced is glycosylated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00570892
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  • 195
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    Retinoids and a retinoic acid receptor differentially modulate thymosinβ 10 gene expression in transfected neuroblastoma cells (1992)
    Springer
    Cellular and molecular neurobiology 12 (1992), S. 45-58 
    Details
    ISSN: 1573-6830
    Keywords: retinoid ; thymosin ; neuroblastoma ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. Investigations have demonstrated that the gene encoding thymosinβ 10 (a 43-amino acid member of a family of related proteins originally described in the rat immune system) is a target for morphogenic retinoids in both human and rat neuroblastoma cells. 2. Structure-activity studies revealed that the stimulatory actions of retinoids upon the thymosinβ 10 gene reflect the differing affinities of retinoid analogues for a retinoic acid receptor. 3. To examine further the possibility that the trophic actions of retinoic acid upon expression of the thymosinβ 10 gene involved retinoid receptors, neuroblastoma cells were transiently transfected with an expression vector encoding the nuclear retinoic acid receptor (α) protein. 4. Northern blot and slot-blot analyses revealed that neuronal cells overexpressing RARα-mRNA exhibited an enhanced sensitivity to exogenous and endogeneous retinoic acid in terms of thymosinβ 10 mRNA. Although the RAR-α gene was expressed (at low levels) a priori in these neuroblastoma cells, retinoic acid (2 × 10−7 M for 3 days) slightly stimulated RAR-α-mRNA accumulation. 5. Collectively, these findings indicate the the retinoic acid receptor (α) is regulated by retinoid acid and that the developmentally regulated, retinoidresponsive thymosinβ 10 gene is a target for this nuclear transcription factor in cells derived from the neural crest.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00711638
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  • 196
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    Differential upregulation of PLP and MAG genes in C6 glioma cells by N2A neuroblastoma conditioned medium (1992)
    Springer
    Metabolic brain disease 7 (1992), S. 147-156 
    Details
    ISSN: 1573-7365
    Keywords: myelin-associated glycoprotein (MAG) ; proteolipid protein (PLP) ; gene expression ; C6 glioma ; N2A neuroblastoma
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The effect of factors released from N2A neuroblastoma cells on the expression of myelin protein genes in glioma C6 cells, i.e., proteolipid protein (PLP) and myelin-associated glycoprotein (MAG), was studied. Both cells lines were propagated in serum-free DMEM-F10 (1:1) medium. The addition of 50% N2A conditioned medium (N2ACM) stimulated the proliferation of C6 cells by approximately 4.5 fold as compared to control cells. The N2ACM-treated cells formed aggregates indicating increased cell-cell affinity. The exposure of C6 cells to N2ACM transiently stimulated the expression of both the MAG-specific and the PLP-specific messages up to eight and four fold over the control values, respectively. The maximal upregulation of the PLP gene occurred two days after N2ACM administration and preceded that of the MAG gene by two days. The effect of N2ACM was dose-dependent in the range of 12.5 to 50 %. The secretion of N2A paracrine factors that stimulated the myelin gene expression was also time-dependent. The optimal conditioning time for the release of the PLP gene-stimulating activity was one day, while the maximal MAG gene-stimulating activity was found in the medium conditioned for 3 days. This cellular system may provide a convenient model for studies on trophic neuronal-glial interaction. Furthermore, the results indicate a difference in the regulatory mechanisms between the PLP and the MAG genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01000160
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  • 197
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    Modulation of gene expression as a biomarker in colon (1992)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 50 (1992), S. 151-157 
    Details
    ISSN: 0730-2312
    Keywords: calcium ; chemoprevention ; colorectal cancer ; fiber ; gene expression ; genetic risk ; intermediate biomarker ; mitochondria ; short-chain fatty acids ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Computer-driven scanning and image processing methodology has demonstrated that genetic inheritance of risk for colorectal cancer in familial polyposis (FAP) and hereditary non-polyposis colorectal cancer (HNPCC) families is associated with highly pleiotropic effects on patterns of gene expression in the flat colonic mucosa. The mitochondrial (mt) gene encoding subunit 3 of cytochrome oxidase (COXIII) is one of a panel of cloned sequences which characterize genetic risk. Expression of COXIII decreased in progression of, and risk for, colonic tumors in vivo. Further, metabolizable, unbranched, short-chain fatty acids (SCFAs) elevated expression of mtCOXIII, as well as mtCOXI, in HT29 cells and also elevated mtCOX enzymatic activity. However, expression of nuclear encoded COX subunits were unaffected. These changes may be related to documented alterations in mitochondria structure and function in transformed colonic epithelial cells.SCFAs produced when colonic microflora causes fermentation of fiber are the principle energy source for normal colonic epithelial cells; SCFAs also induce a more differentiated phenotype both in vitro and in vivo. Therefore, a mechanistic link may exist between molecular events in inherited risk and a dietary factor (fiber) which may modulate such risk.In a preliminary intervention trial in collaboration with M. Lipkin, high risk HNPCC patients received daily supplements of 1500 mg CaCO3 per day, which may be protective for development of colorectal tumors. Elevations in COXIII expression were seen in 7 of 12 patients within the first 7 months, followed by complex changes in expression of this sequence. © 1992 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcb.240501126
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  • 198
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    Stage-specific protein and mRNA synthesis during morphogenesis of the polyembryonic parasitoid Copidosoma floridanum (Ashmead) (Hymenoptera: Encyrtidae) (1992)
    New York, NY [u.a.] : Wiley-Blackwell
    Archives of Insect Biochemistry and Physiology 19 (1992), S. 81-92 
    Details
    ISSN: 0739-4462
    Keywords: development ; embryogenesis ; gene expression ; Trichoplusia ni ; Lepidoptera ; Noctuidae ; Chemistry ; Food Science, Agricultural, Medicinal and Pharmaceutical Chemistry
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Changes in protein and mRNA synthesis during embryonic morphogenesis of the polyembryonic parasitoid Copidosoma floridanum were characterized by 1- and 2-dimensional gel electrophoresis and quantified by scanning densitometry. Analysis of protein synthesis at different developmental stages indicated that embryonic molecular changes occurred at 36 h of the host's fourth stadium. These changes included decreased expression of 51 and 104 kD (D) proteins and increased expression of 22, 22.5, and 24 kD (D) proteins. Similarly, analysis of in vitro translation products indicated increased transcription of mRNA encoding 16 and 49 kD proteins in embryos dissected at 36 h of the host's fourth stadium. The stage-specific changes in transcription and translation corresponded to the blastula stage of embryonic development and arose several hours before the initiation of embryonic morphogenic events associated with larval pattern formation. Application of the JH analogue methoprene at times that block morphogenesis but not blastula formation did not block the stage-specific synthesis of any proteins. In contrast, neck ligation of hosts at times that block blastula formation and morphogenesis inhibited the synthesis of the 24 kD protein that was normally expressed at the blastula stage.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/arch.940190202
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  • 199
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    Transgenic regulation in laboratory animals (1991)
    Springer
    Cellular and molecular life sciences 47 (1991), S. 866-877 
    Details
    ISSN: 1420-9071
    Keywords: Transgenic mice ; microinjection ; recombinant DNA ; gene expression ; transcription factors ; chromatin ; homologous recombination ; episomal maintenance ; embryonic stem cells ; germ line ; position-effect ; mosaicism ; globin genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract This chapter is an attempt to summarize some commonly accepted and some more subjective opinions about the regulation of transgene expression in laboratory animals. After a short historical introduction, I present some general notions regarding gene structure/function. The spotlight shifts then to the description of the most popular techniques for gene transfer, including the targeted gene replacement. The different approaches are briefly discussed in terms of intrinsic advantages and limitations regarding gene expression patterns. Furthermore, the role of enhancers, promoters and othercis-acting elements such as silencers and dominant control regions as well as their involvement in the chromatin on-off state are discussed on the basis of a specific example studied in our laboratory. The review concludes by presenting recent results and the new perspectives opening in the field of ‘surrogate’ (also called ‘reversed’) genetics. Some problems which remain to be solved both at the technical as well as at the social-ethical level are also briefly presented.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01929876
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  • 200
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    Production of pharmaceutical proteins in milk (1991)
    Springer
    Cellular and molecular life sciences 47 (1991), S. 905-912 
    Details
    ISSN: 1420-9071
    Keywords: Gene transfer ; gene modification ; gene expression ; livestock ; transgenic animal ; pharmaceutical proteins ; milk composition
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract There is every reason to expect that it will be possible within the next few years to begin to use farm animals to produce large quantities of some of the human proteins that are needed for the treatment of disease. Revolutionary new opportunities for the production of novel proteins in milk have been created by the development of methods for gene transfer. Exploitation of these opportunities depends upon selection and cloning of milk protein genes and identification of the sequences that govern tissue specific hormonally induced expression in the mammary gland. Studies with three genes, ovine β-lactoglobulin, rat β-casein and whey acidic protein of rat and mouse, suggest that they may all meet this requirement. Fragments of the ovine β-lactoglobulin, murine whey acidic protein and rabbit β-casein genes have directed production of novel proteins in the milk of transgenic mice, sheep, rabbits and pigs. The proteins were biologically active and usually co-migrated with authentic proteins. In early experiments, protein concentration was low, but our recent observations suggest that fusion genes containing genomic clones direct production of concentrations of protein that are suitable for commercial exploitation. In the longer term, two approaches may offer the potential of more reliable expression. Control elements capable of directing expression that is independent of site of insertion of the gene, but dependent on the number of copies of the gene, have been identified for a small number of genes. The availability of such elements for the milk protein genes would increase the reliability of gene expression considerably. Alternatively, targeted mutation of genes may allow the insertion of coding sequences within an existing gene so avoiding position effects.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01929881
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