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  • 2020-2023
  • 1985-1989  (114)
  • 1955-1959  (8)
  • 1950-1954
  • gene expression
  • protoplasts
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Cellular and molecular life sciences 45 (1989), S. 526-535 
    ISSN: 1420-9071
    Keywords: Interferon ; cytokines ; interleukins ; gene expression ; transcription ; autoimmune ; disease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Specific interferon genes are transcribed at low levels in the spleen, liver, and peripheral blood leukocytes of normal individuals in the apparent absence of virus infection while other interferon genes remain unexpressed in the same tissues. In contrast, the genes of cytokines such as IL-1, IL-6 and TNF are expressed at relatively high levels in the organs of normal individuals. The level of expression of the IL-1, IL-6 and TNF genes is markedly reduced in the livers of patients with autoimmune liver disease compared to the level of expression in the liver of normal individuals, whereas the expression of interferon genes is similar in both normal and diseased liver, suggesting that a defect in the expression of specific cytokines is associated with severe liver disease.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-0832
    Keywords: dermatophyte ; Microsporum gypseum ; Novozyme ; protoplasts ; regeneration ; viability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract Factors affecting high yields, regeneration frequencies, and viability of protoplasts from clonal cultures of Microsporum gypseum were investigated. Maximum yields of protoplasts were obtained after 6 hrs digestion of 2–4 days old mycelium with Novozyme 234 using CaCl2 (0.4 M) as an osmotic stabilizer and glycine + HCl (pH 4.5) as the buffer system. Mercaptoethanol + dithiothreitol (0.01 M) proved to be the best pretreatment of mycelium prior to digestion with enzyme. A regeneration frequency of 94.4% was obtained using the top agar method with complete medium (pH 6.5) containing 0.5% agar and 0.4 M CaCl2 as an osmoticum. Colonies from regenerated protoplasts on medium containing CaCl2 were pigmented and completely powdery with high sporulation. Protoplast viability was studied in osmotic stabilizer supplemented with glucose or glutamine. After 24 hrs, glucose (2%) and glutamine (2%) enhanced protoplast viability by 22% and 23%, respectively. Protein synthesis, as measured by 3H-lysine uptake, matched the viability profile determined by fluorescence microscopy.
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Methods in cell science 12 (1989), S. 127-133 
    ISSN: 1573-0603
    Keywords: polyethylene glycol ; transformation ; protoplasts ; Nicotiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Polyethylene glycol can be used to induce DNA uptake into plant protoplasts. Procedures for isolation, culture and transformation ofN. tabacum protoplasts are described and can be adapted for other dicot and monocot species. Criteria for proof of transformation are discussed.
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell reports 8 (1989), S. 313-316 
    ISSN: 1432-203X
    Keywords: Zea mays L. ; supersweet corn ; protoplasts ; haploids ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Plants were regenerated from maize (Zea mays L.) protoplasts isolated from embryogenic cell suspensions. The donor maize suspension cultures were established from friable callus initiated from microspores of a commercial supersweet hybrid (sh2sh2). The frequency of cell colony formation was higher when protoplasts were cultured on feeder layers of maize cells as compared with a liquid thin layer method. It was demonstrated that haploid and dihaploid soil-grown plants can be regenerated from maize protoplasts isolated from haploid cell cultures.
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  • 5
    ISSN: 1432-203X
    Keywords: sunflower ; protoplasts ; direct gene transfer
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Helianthus annuus protoplasts were transformed with the plasmid pCaMVNEO (Frommet al. 1986) conferring kanamycin resistance to plant. Transformed calli were selected with a frequency of 4 calli for 106 treated protoplasts. DNA was extracted from kanamycin resistant calli. Analysis of this DNA shows the presence of the NPTII gene.
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  • 6
    ISSN: 1573-4927
    Keywords: mouse ; salivary protein ; lacrimal protein ; gene expression ; Spt locus ; multigene family
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The murine submandibular gland (SMG) produces a novel class of highly acidic salivary proteins encoded by one or more highly abundant mRNA transcripts. In inbred mice, these transcripts are encoded by members of a multigene family comprising approximately 8–12 homologues. Most, and probably all, of these homologues are clustered at a new locus near belted (bt) on chromosome 15, which we designateSpt (salivary protein). Although physically closely linked,Spt genes differ in their patterns of expression both in strains of mice and in their tissues. One gene,Spt-1, is expressed at high levels in the SMG of all inbred strains examined. This gene is also expressed at significant levels in the lacrimal gland. A second gene,Spt-2, appears to be present as a single copy in some strains and as two copies in others. This gene is expressed at high levels only in the SMG of those strains carrying two copies, andSpt-2 mRNA is not detectable in the SMG of strains carrying only one copy. In contrast toSpt-1, theSpt-2 gene is not expressed at detectable levels in the lacrimal gland.
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  • 7
    ISSN: 1573-4927
    Keywords: mouse ; salivary protein ; lacrimal protein ; gene expression ; Spt locus ; multigene family
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The murine submandibular gland (SMG) produces a novel class of highly acidic salivary proteins encoded by one or more highly abundant mRNA transcripts. In inbred mice, these transcripts are encoded by members of a multigene family comprising approximately 8–12 homologues. Most, and probably all, of these homologues are clustered at a new locus near belted (bt) on chromosome 15, which we designateSpt (salivary protein). Although physically closely linked,Spt genes differ in their patterns of expression both in strains of mice and in their tissues. One gene,Spt-1, is expressed at high levels in the SMG of all inbred strains examined. This gene is also expressed at significant levels in the lacrimal gland. A second gene,Spt-2, appears to be present as a single copy in some strains and as two copies in others. This gene is expressed at high levels only in the SMG of those strains carrying two copies, andSpt-2 mRNA is not detectable in the SMG of strains carrying only one copy. In contrast toSpt-1, theSpt-2 gene is not expressed at detectable levels in the lacrimal gland.
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  • 8
    ISSN: 1573-4919
    Keywords: complement ; factor B ; gene expression ; interferon-ψ ; interleukin-1
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract The serine protease complement factor B (Bf), an acute phase plasma protein, is a component of the alternative pathway of complement activation. Previous studies revealed that several cytokines including IFN-γ and IL-1 are involved in mediating acute phase Bf expression. To determine the molecular details of Bf expression we isolated, sequenced and characterized the 5′ flanking regions of the human and murine Bf genes. In both species the Bf transcriptional start site in liver was located 〈400 by 3′ to the polyadenylation site of the upstream C2 gene. This upstream intergenic region contained 〉65% nucleotide homology between species. Within this region, an IRS and three heat shock consensus elements were found in the murine sequence in an identical position to that of the human. To examine the functional details of Bf expression, a series of mouse and human Bf promoter - chloramphenicol acetyltransferase (CAT) chimeric gene constructs were transfected into mouse L or human HepG2 cells. Analysis of expression of these fusion gene constructs revealed that 1) cis-acting DNA sequences identified, at least in part, in the 3′ untranslated region of the C2 gene (within the 400 by upstream of the Bf cap site) mediate responsiveness to IL-1 and IFN-γ, 2) the responsiveness to each mediator appears to be conferred by separate upstream regions similar in position and homologous in man and mouse, and 3) the IL-1 responsive region in both species appears to have the characteristics of an enhancer element. The results of this analysis suggest a selective pressure to conserve the intergenic sequence between C2 and Bf genes and that further studies of these sequences will be useful in elucidating mechanisms controlling the acute phase response.
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  • 9
    ISSN: 1573-5028
    Keywords: differential cDNA screening ; gene expression ; hybrid-select in vitro translation ; nuclear run-off transcription ; RNA blot hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cDNA library from cultured parsley (Petroselinum crispum) cells was differentially screened using labeled run-off transcripts derived from nucleic of elicitor-treated and untreated cells. This resulted in the isolation of 18 independent cDNA families representing putative defense-related genes. All genes are rapidly and transiently activated after elicitor application, but the time courses of transcriptional activity exhibit considerable variations, indicating differences in the mechanisms of gene regulation.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Bioscience reports 9 (1989), S. 157-188 
    ISSN: 1573-4935
    Keywords: matrix hypothesis ; morphogenesis ; protodynamics ; growth control ; DNA ; genome organisation ; gene expression ; gene regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A theoretical concept is proposed, in order to explain some enigmatic aspects of cellular and molecular biology of eukaryotic organisms. Among these are the C-value paradox of DNA redundancy, the correlation of DNA content and cell size, the disruption of genes at DNA level, the “Chromosome field” data of Lima de Faria (Hereditas 93∶1, 1980), the “quantal mitosis” proposition of Holtzeret al. (Curr. Top. Dev. Biol. 7∶229 1972), the inheritance of morphological patterns, the relations of DNA and chromosome organisation to cellular structure and function, the molecular basis of speciation, etc. The basic proposition of the “Unified Matrix Hypothesis” is that the nuclear DNA has a direct morphogenic function, in addition to its coding function in protein synthesis. This additional genetic information is thought to be largely contained in the non-protein coding transcribed DNA, and in the untranscribed part of the genome.
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  • 11
    ISSN: 1573-5028
    Keywords: acyl carrier protein ; fatty acid synthesis ; gene expression ; gene fusion ; protein A ; spinach
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A synthetic gene encoding spinach acyl carrier protein I (ACP-I) was fused to a gene encoding the Fc-binding portion of staphylococcal protein A. This gene fusion, under the control of the λPR promoter, was expressed at high levels in Escherichia coli producing a 42 kDa fusion protein. This fusion protein was phosphopantethenylated in E. coli. In vitro the ACP portion of the fusion protein was able to participate in acyl ACP synthetase reactions, plant malonyl-CoA:ACP transacylase (MCT) reactions, and plant fatty acid synthetase (FAS) reactions. Inhibitory effects of high ACP concentrations on in vitro plant FAS were observed with the unfused ACP-1 but not with the fusion protein. As with unfused ACP-I, the fusion protein was a poor substrate for E. coli FAS reactions. When injected into rabbits, the fusion protein was also able to generate antiserum to spinach ACP-I.
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  • 12
    ISSN: 1573-5028
    Keywords: cDNA sequence ; gene expression ; glutamine synthetase ; legume-Rhizobium symbiosis ; nitrogen metabolism ; Phaseolus vulgaris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We report the sequence of an essentially full-length glutamine synthetase (GS) cDNA clone (pcGS-γ1) isolated from a root nodule library ofPhaseolus vulgaris L. The polypeptide encoded by this cDNA has been producedin vitro by transcription/translation and shown to co-migrate on two-dimensional gels with the previously identified major cytosolic GS polypeptide (γ) of nodules. Two previously identified GS cDNA clones, pR-2 and pR-1 (see Gebhardtet al., EMBO J 5: 1429–1435, 1986) have similarly been shown to encode the α and β cytosolic GS polypeptides respectively. An RNase protection technique has been used to analyse specifically and quantitatively the abundance of mRNA related to these three GS cDNAs and to the cDNA (pcGS-δ1) encoding the chloroplast-located GS, during nodulation. Differences in the abundances of these mRNAs at different times suggest that they are not coordinately regulated. Moreover, using this technique mRNA specifically related to pcGS-γ1 was found at high levels in nodules but not in roots or leaves. Surprisingly the expression of this gene is not nodule-specific as previously suggested, as its mRNA was also detected, but at lower levels, in stems, petioles and in green cotyledons. By comparison, mRNA related to a leghaemoglobin gene was detected only in nodules. Comparisons of the relative abundances of the pcGS-γ1 mRNA and the γ polypeptide in different organs and at different stages during nodulation, suggest that the appearance of the γ polypeptide is largely under transcriptional control.
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  • 13
    ISSN: 1573-5028
    Keywords: aleurone ; barley ; protoplasts ; transient expression ; wheat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Methods have been developed for the isolation of aleurone protoplasts from developing caryopses of Hordeum vulgare and Triticum aestivum in order to study transient expression of introduced genes. Chimaeric gene constructs were introduced into aleurone protoplasts by polyethylene glycol (PEG). Transient expression directed by the 35S promoter from cauliflower mosaic virus (CaMV) of the reporter gene encoding chloramphenicol acetyl transferase (CAT) was detected in aleurone protoplasts from developing barley and wheat grains. Using a similar construct, CAT activity increased when the alcohol dehydrogenase intron 1 fragment from maize was ligated between the 35S promoter and the CAT coding region. The demonstration of transient expression in protoplasts from developing aleurone layers indicates that they may be useful for investigating tissue and developmental control of genes coding for cereal seed proteins.
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  • 14
    ISSN: 1573-5028
    Keywords: antisense RNA ; β-glucuronidase ; tobacco ; transgenic plants ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Antisense RNA was used to specifically inhibit the expression of a GUS gene introduced in a transgenic plant. A tobacco transformant containing a single intact copy of the GUS gene and showing relatively high constitutive levels of GUS activity (GUS+) was re-transformed with an Agrobacterium Ti-derived binary vector containing an antisense version of this reporter gene. The sense and antisense GUS genes were each under the regulation of the CaMV 35S promoter. Re-transformed plants contained 1–5 copies of the antisense construct and all showed a greater than 90% reduction in GUS activity relative to the original GUS+ plant. This reduction in GUS activity correlated closely with the levels of GUS enzyme and steady state GUS mRNA observed in these plants. The relatively low levels of sense and antisense GUS transcripts found in the re-transformed plants may indicate a rapid degradation of the RNA:RNA duplex in the cell.
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  • 15
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 13 (1989), S. 503-511 
    ISSN: 1573-5028
    Keywords: electroporation ; patatin genes, potato ; protoplasts ; transient gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Electroporation was used to evaluate parameters important in transient gene expression in potato protoplasts. The protoplasts were from leaves of wild potato Solanum brevidens, and from leaves, tubers and suspension cells of cultivated Solanum tuberosum cv. Désirée. Reporter enzyme activity, chloramphenicol acetyl transferase (CAT) under the control of the cauliflower mosaic virus (CaMV) 35S promoter, depended on the field strength and the pulse duration used for electroporation. Using field pulses of 85 ms duration, the optimum field strengths for maximum CAT activity were: S. brevidens mesophyll protoplasts −250 V/cm; Désirée mesophyll protoplasts −225 V/cm; Désirée suspension culture protoplasts −225 V/cm; and Désirée tuber protoplasts −150 V/cm. The optimum field strengths correlated inversely with the size of the protoplasts electroporated; this is consistent with biophysical theory. In time courses, maximum CAT activity (in Désirée mesophyll protoplasts) occurred 36–48 h after electroporation. Examination at optimised conditions of a chimaeric gene consisting of a class II patatin promoter linked to the β-glucuronidase (gus) gene, showed expression (at DNA concentrations between 0–10 pmol/ml) comparable to the CaMV 35S promoter in both tuber and mesophyll protoplasts. At higher DNA concentrations (20–30 pmol/ml) the patatin promoter directed 4–5 times higher levels of gus expression. Implications and potential contributions towards studying gene expression, in particular of homologous genes in potato, are discussed.
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  • 16
    ISSN: 1573-5028
    Keywords: deformation factor ; gene expression ; pea ; Rhizobium leguminosarum ; root hairs
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The mRNA population in pea root hairs was characterized by means of in vitro translation of total root hair RNA followed by 2-dimensional gel electrophoresis of the translation products. Root hairs contain several mRNAs not detectable in total RNA preparations from roots. Most of these root hair-specific mRNAs occur in elongating root hairs at higher levels than in mature root hairs. The expression of some genes in pea root hairs is typically affected by inoculation with Rhizobium leguminosarum. One gene, encoding RH-42, is specifically induced while the expression of another gene, encoding RH-44, is markedly enhanced. Using R. leguminosarum mutants it was shown that the nodC gene is required for the induction and enhancement of expression of the RH-42 and RH-44 genes, respectively, while the Rhizobium chromosomal gene pss1, involved in exopolysaccharide synthesis, is not essential. After induction of the nod genes with apigenin the bacteria excrete into the culture medium a factor that causes root hair deformation. This deformation factor stimulates the expression of the RH-44 gene but does not induce the expression of the gene encoding RH-42.
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  • 17
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 12 (1989), S. 295-299 
    ISSN: 1573-5028
    Keywords: gene expression ; multiple transcripts ; phytochrome ; Pisum sativum ; red/far-red reversible effect
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The reversible effect of red and far-red light on the level of two RNA transcripts of the single-copy phytochrome gene in pea was investigated using the primer extension assay. In dark-grown seedlings, a brief irradiation with red light markedly reduced the level of one of the phytochrome transcripts, RNA1. This red light effect was reversed by subsequent irradiation with far-red light. The other phytochrome transcript, RNA2, was only slightly influenced by light. In light-grown seedlings, a brief irradiation with far-red light increased the content of both RNA1 and RNA2 when the seedlings were transferred from light to darkness. This far-red light effect was reversible by subsequent red light irradiation.
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  • 18
    ISSN: 1573-5028
    Keywords: field inversion gel electrophoresis ; DNA isolation ; protoplasts ; nuclear DNA ; chloroplast DNA ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Schwartz and Cantor technique for releasing and fractionating megabase-sized DNA from agarose-embedded cells is beginning to bridge the gap in resoluation between classical genetics and current molecular DNA techniques, particularly in mammalian systems. As yet no conditions have been described for preparing plant DNA that is of sufficient length to allow similar long-range restriction mapping experiments in plant systems. In this report, we describe the application of the Schwartz and Cantor technique for preparing high molecular weight DNA from embedded tomato leaf protoplasts, as well as conditions for generating and fractionating large restriction fragments, by field inversion gel electrophoresis (FIGE). The bulk of DNA released from lysed protoplasts was at least 2 Mb in size and amenable to restriction digestion as shown by hybridizing Southern blots with, among others, a probe for the Adh-2 gene of tomato. Restriction fragments as large as 700 kb were detected. Chloroplast DNA is isolated intact, amenable to restriction analysis and, in its native form, not mobile in FIGE.
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  • 19
    ISSN: 1573-5028
    Keywords: C4 photosynthesis ; gene structure ; gene expression ; genetic variation ; silent substitution ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have determined the structure of the maize (Zea mays L. subsp.mays line B73) nuclear gene encoding the phosphoenolpyruvate (PEP) carboxylase isozyme involved in C4 photosynthesis. The gene is 5.3 kb long and has ten exons that range in size from 85 to 999 bp. The nine introns vary from 97 to 872 bp. The sequence of 663 bp of 5′-flanking and 205 bp of 3′-flanking DNA is reported along with the entire gene sequence. Several short repetitive sequences were found in the 5′-flanking DNA that have characteristics similar to elements important in the light regulation of pea genes encoding the small subunit of ribulose 1,5-bisphosphate carboxylase. In addition, some 5′-flanking sequence similarities were found in a comparison with other light-regulated genes from maize and wheat. The level of DNA sequence variation among different PEP carboxylase alleles is similar to the allelic variation observed for several other maize nuclear genes. The data suggest modern maize variaties have retained much of the genetic variation present in their ancestral forms. Finally, accumulation of transcripts encoding the PEP carboxylase isozyme involved in C4 photosynthesis is quite high in several structures besides leaves, including inner leaf sheaths, tassels and husks. This indicates that expression of this gene is not leaf-specific and may not necessarily be coupled to the development of Kranz anatomy.
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  • 20
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 12 (1989), S. 673-682 
    ISSN: 1573-5028
    Keywords: amylase inhibitor ; barley ; cDNA sequence ; gene expression ; hormonal regulation ; protease inhibitor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have cloned and sequenced a full-length cDNA from barley (Hordeum vulgare L.) seeds encoding the bifunctional α-amylase/subtilisin inhibitor (BASI). The nucleotide sequence predicts an open reading frame coding for a protein of 203 amino acids. The first 22 amino acids exhibit the sequence characteristic of a signal peptide, as found in several other plant protease inhibitors. Northern blot hybridization experiments indicate that BASI mRNA accumulation is strictly tissue-specific and is developmentally programmed. BASI mRNA transcripts were only identified in 1) developing starchy endosperm tissue from 14 days after flowering and 2) aleurone tissue of germinating seeds. In this latter tissue, BASI mRNA accumulation is enhanced by abscisic acid and abolished by gibberellic acid. Expression of BASI mRNA was also studied in the lys 3a high-lysine barley mutants Risø No. 1508 and Piggy. These high-lysine barleys show 2–4-fold higher levels as well as prolonged accumulation of BASI mRNA compared to the normal motherline Bomi. This correlates with the increased deposition of BASI protein in lys 3a barley mutants. Genomic blot analysis of barley DNA suggests that there are one or two BASI structural genes per haploid genome. Possible roles of BASI as part of a defence mechanism against precocious germination and pathogens are discussed.
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  • 21
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 13 (1989), S. 151-161 
    ISSN: 1573-5028
    Keywords: β-glucuronidase (gusA) gene ; maize ; protoplasts ; stable co-transformation ; transformation ; Zea mays L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An efficient co-transformation protocol using polyethylene glycol was developed for Zea mays L. (cv. A188 × BMS) protoplasts isolated from suspension culture cells. Co-transformation was accomplished by using plasmid constructions containing β-glucuronidase (gusA) or neomycin phosphotransferase (neo) gene coding sequences; both were under control of the CaMV 35S promoter. Protoplast culture and transformation conditions were optimized to assure efficient recovery of transformed cells. The overall efficiency of transformation was 1 × 10−4 (calculated per viable protoplast plated). Among kanamycin-resistant lines, 50% showed a high level of GUS activity (above one unit). Southern blot hybridization confirmed the presence of numerous gusA and neo coding sequences in the maize genome. In two analyzed lines, integrated sequences appeared to be organized in tandem head-to-tail repeats. Results also indicated that the integrated sequences were partially methylated.
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  • 22
    ISSN: 1573-5028
    Keywords: cadmium ; Datura innoxia ; gene expression ; heat shock ; tolerance
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of Cd on gene expression in suspension cultures of twoDatura innoxia cell lines with differing Cd tolerance was studied.In vivo labeling experiments using [3H] leucine showed that Cd induced the synthesis of a similar range of proteins in both cell lines at a concentration which will kill the sensitive but not the tolerant cells. Corresponding changes in levels of translatable mRNA were also observed. The induction of the synthesis of proteins by Cd was transient since Cd-tolerant cells growing continuously in 250 μM CdCl2 contained a similar set ofin vitro translation products to cells growing in the absence of Cd. Although Cd had a similar effect on gene expression in both cell lines, Cd-tolerant cells possess two abundant mRNAs which are constitutively produced. These mRNAs encode proteins of low molecular weight (about 11 kDa) and are either absent or present at a low level in Cd-sensitive cells. The functions of these proteins are not known but they may be involved in the tolerance mechanism. Two-dimensional gel electrophoresis ofin vitro translation products showed that many of the Cd-induced proteins are also induced by heat shock. A 42°C heat shock resulted in agreater range and more intense induction of translatable mRNAs than 4 h exposure to 250 μM CdCl2. However a subset of mRNAs were induced specifically by Cd while other mRNAs were heat shock-specific. There was no difference in the ability of the two cell lines to tolerate heat shock. This was also reflected by the same pattern of major proteins induced by heat shock in the two cell lines.
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  • 23
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 12 (1989), S. 667-672 
    ISSN: 1573-5028
    Keywords: ferredoxin ; gene expression ; heterocyst ; Anabaena 7120 ; Escherichia coli
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The genes for ferredoxin from heterocysts (fdx H) and vegetative cells (pet F) of Anabaena sp. strain 7120 were subcloned into plasmid pUC 18/19. Both genes were expressed in Escherichia coli at high levels (≈10% of total protein). Pet F could be expressed from its own promoter. The ferredoxins were correctly assembled to the holoprotein. Heterocyst ferredoxin was purified from E. coli extracts on a large scale. Its biochemical and biophysical properties were identical to those of the authentic ferredoxin, isolated from Anabaena heterocysts.
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  • 24
    ISSN: 1573-5028
    Keywords: cDNA ; gene expression ; hydroxypyruvate reductase ; light regulation ; peroxisomal enzyme
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A full-length cDNA encoding NADH-dependent hydroxypyruvate reductase (HPR), a photorespiratory enzyme localized in leaf peroxisomes, was isolated from a λgt11 cDNA library made by reverse transcription of poly(A)+ RNA from cucumber cotyledons. In vitro transcription and translation of this clone yielded a major polypeptide which was identical in size, 43 kDA, to the product of in vitro translation of cotyledonary poly(A)+ RNA and subsequent immunoprecipitation with HPR antiserum. Escherichia coli cultures transformed with a plasmid construct containing the cDNA insert were induced to express HPR enzyme activity. RNA blot analysis showed that HPR transcript levels rise significantly in the first eight days of light-grown seedling development. This closely resembles the pattern seen for HPR-specific translatable mRNA. DNA blot analysis indicated that a single HPR gene is likely present per haploid genome. Nucleotide sequence analysis revealed an open reading frame of 1146 bases which encodes a polypeptide with a calculated molecular weight of 41.7 kDa. The derived amino acid sequence from this open reading frame is 26% identical and 50% similar to the amino acid sequence of the E. coli enzyme phosphoglycerate dehydrogenase, which catalyzes a similar reaction and functions in a related pathway. Statistical analyses show that this similarity is significant (z〉10). The derived amino acid sequence for HPR also contains the characteristics of an NAD-binding domain.
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  • 25
    Electronic Resource
    Electronic Resource
    Springer
    Bulletin of experimental biology and medicine 108 (1989), S. 999-1001 
    ISSN: 1573-8221
    Keywords: brain ; gene expression ; mRNA ; transplantable hepatomas
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
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  • 26
    Electronic Resource
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    Springer
    Cellular and molecular neurobiology 9 (1989), S. 281-294 
    ISSN: 1573-6830
    Keywords: cholecystokinin ; mammillary region ; development ; gene expression ; hybridization histochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary 1. The ontogeny and adult distribution of neurons containing cholecystokinin (CCK) mRNA in the premammillary and mammillary nuclei and supramammillary region of the rat brain were studied using hybridization histochemistry. 2. The earliest detection of CCK mRNA in the mammillary region was on E14, followed by a marked increase in transcript levels during the next 4 days, a time during which neurons in this region still divide. During the first 2 weeks of life, few changes in the levels of CCK transcripts were seen, and an adult-like pattern of expression was seen on the twenty-first day of life. 3. Low levels of transcripts were present in numerous neurons located in all divisions of the medial nucleus and in the posterior nucleus known to project ipsilaterally to the anteroventral and anteromedial thalamic nuclei. In contrast, none of the neurons in the lateral nucleus (projecting bilaterally to the anterodorsal thalamic nucleus) had detectable transcripts. 4. Many neurons in the supramammillary nucleus had low, moderate, or high levels of transcripts. Some nearby nuclei (such as the dorsal premammillary nucleus) had smaller numbers of neurons with low levels of CCK mRNA, whereas others (such as the ventral premammillary nucleus) had none.
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  • 27
    ISSN: 1573-5060
    Keywords: Lycopersicon ; tomato ; haploids ; chromosomal instability ; chloroplast number ; callus culture ; protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary The effect of the tissue culture system, the genotype and the ploidy level of the plant material used as explant source on the stability of the ploidy level of plants regenerated fromcell and tissue cultures of tomato was investigated. In addition the use of the chloroplast number in guard cells as a measure for ploidy level was evaluated. Haploids of tomato were very instable, which instability was observed already in somatic root tip and leaf cells. The number of regenerated plants that retained the original ploidy level differed significantly between the tested haploids. The plants that were regenerated from leaf explants of diploids were predominantly diploid in contrast to the plants regenerated from established callus cultures and protoplast where the majority was tetraploid.
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  • 28
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    Plant cell, tissue and organ culture 19 (1989), S. 103-111 
    ISSN: 1573-5044
    Keywords: amino acids ; ammonium ; growth factors ; light ; mannitol ; nitrate ; organogenesis ; polyamines ; potato ; protoplasts ; Solanum tuberosum ; sucrose ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The influence of several media components and environmental factors on shoot formation in protoplast-derived calli of Solanum tuberosum (a Rosamunda cross) were studied. Low sucrose concentration (3–15 mM) was beneficial for optimal shoot induction. Several concentrations of NO3 - and NH4 + were suitable for shoot induction as long as the concentration of NO3 - was about twice the concentration of NH4 + or higher. No stimulatory effect of glutamine, proline, putrescine, spermidine, spermine or adenine sulphate at 0.5 and 2 mM were found. White light promoted shoot induction compared with red and blue light or darkness. The intensity of light was shown to be a critical factor for good shoot induction. Lower light intensity (30 μE m-2 s-1) resulted in doubling of the number of calli producing shoots compared with higher (110 μE m-2 s-1) light intensity. A temperature of 20°C promoted shoot regeneration compared to 25°C. Based on these results improved conditions for regeneration of S. tuberosum are suggested, and shown to enhance shoot formation in five other genotypes tested.
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  • 29
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    Plant cell, tissue and organ culture 19 (1989), S. 113-127 
    ISSN: 1573-5044
    Keywords: somatic embryogenesis ; plant regeneration ; protoplasts ; Trifolium pratense ; red clover ; protoclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts are useful for subcellular studies, in vitro selection, somatic hybridization and transformation. Whole plant regeneration from protoplasts is a prerequisite to producing altered crop plants using these methods. Whole plant regeneration was achieved from leaf- and suspension culture-derived protoplasts of T. pratense. Regeneration was most dependent upon identifying genotypes with genetic capacity to regenerate. Additional factors that were used to select genotypes, but which proved to be less important, were a high rate of cell growth in culture and a high plating efficiency of protoplasts. One genotype was identified which had a regeneration response equivalent to that of T. rubens and which regenerated from both leaf- and suspension culture-derived protoplasts.
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  • 30
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    Methods in cell science 12 (1989), S. 139-144 
    ISSN: 1573-0603
    Keywords: microinjection ; genetic transformation ; protoplasts ; microspores ; Brassica napus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This paper describes a general method suitable for the microinjection ofBrassica napus protoplasts, unicellular microspores, and multicellular microspores. By incorporating components taken from other methods, manual operations frequently involved in the microinjection of plant cells have been simplified and microinjection rates increased. The embedding of cells in agarose provides a simple alternative to the variety of sophisticated immobilization strategies devised for different plant cell types thereby reducing the manipulations often involved in the culture of microinjected cells. Use of an automatic microinjector eliminated the operation of fine control systems on manual injectors; however, precision in sample delivery was reduced. Analyses indicate that transformed tissues can be recovered from microinjected protoplasts and microspores at high frequencies.
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  • 31
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    Methods in cell science 12 (1989), S. 157-161 
    ISSN: 1573-0603
    Keywords: protoplasts ; fusion ; somatic ; hybridization ; cybridization ; asymmetric hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary This report describes several techniques for plant gene transfer by protoplast fusion. These detailed procedures are adaptable to a wide variety of plant species for the transfer of either the entire or partial genomes of nuclear, mitochondrial, and chloroplast origins between species. Detailed procedures should allow researchers to modify and adapt these techniques to suit their own requirements. Effort has been made to describe the protoplast fusion techniques used in the authors' laboratory as comprehensively as possible while citing the many alternatives and modifications that other workers have successfully employed.
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  • 32
    ISSN: 1572-994X
    Keywords: BAL31 nuclease ; CAT assay ; gene expression ; HTLV-I ; R region ; 5′ untranslated sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract We previously demonstrated the activity of the R fragment in the long terminal repeat of human T-cell leukemia virus type I for elevation of the level of gene expression. In this study, the fragment was deleted with BAL31 nuclease to determine its functional domain. Series of the shortened R fragments were linked to the simian virus 40 promoter unit, which regulated expression of a reporter gene. Examination with the R fragments deleted from the 5′ and 3′ ends showed that borders of the functional domain were mapped within nucleotide positions 458 to 473 for the 5′ end and nucleotide positions 559 to 594 for the 3′ end, respectively. Thus we conclude that a 136-base-pair fragment corresponding to the second half of the R region was sufficient to allow elevation of the level of gene expression.
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  • 33
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    Plant cell, tissue and organ culture 19 (1989), S. 213-224 
    ISSN: 1573-5044
    Keywords: Brassica napus ; B. oleracea ; rapid-cycling brassica populations ; protoplasts ; regeneration ; maltose
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts were isolated from aseptic shoot cultures of commercial cultivars ofBrassica napus, B. oleracea andB. campestris, and from the six ‘rapid-cycling brassica species’. Of the rapid-cycling species, onlyB. napus responded well to the culture conditions used; 2% of protoplasts formed calli and up to 5% of calli regenerated shoots. Regeneration was also achieved from commercial cultivars ofB. napus andB. oleracea. For these two species the plating density, time of dilution with fresh medium and the composition of the shoot-inducing medium were all found to have an important influence on the efficiency of plant regeneration. Both responded better to maltose than to sucrose-based media. Under the optimum conditionsB. napus showed a plating efficiency of 7.8% and shooting efficiency of 17%; forB. oleracea the figures were 2% and 56%, respectively.
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  • 34
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    Plant cell, tissue and organ culture 17 (1989), S. 91-100 
    ISSN: 1573-5044
    Keywords: amino acids ; Brassica oleracea ; light ; nitrate ; organogenesis ; polyamines ; protoplasts ; regeneration ; sucrose
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The effect of media components and environmental factors on growth and organogenesis of protoplast-derived calli of curly kale and cabbage were tested. Optimal growth (fresh weight increase of calli, shoots and roots) was found at 60 mM sucrose. Lower sucrose concentrations (3–30 mM) were favourable for shoot formation. Nitrate concentrations from 23 to 100 mM in combination with 8 or 21 mM ammonium were optimal for shoot formation. However, growth was reduced by high (100 mM) nitrate concentration. The effects of various organic nitrogen compounds at 0.5 and 2 mM were tested. Glutamine did not influence shoot formation and barely growth. Proline at 0.5 mM stimulated growth of cabbage calli but decreased growth of curly kale calli, and at 2 mM, proline also inhibited shoot production. Adenine sulphate decreased growth of cabbage calli at 0.5 mM, and at 2 mM shoot production was also reduced. Spermidine and spermine inhibited both growth and differentiation. Putrescine resulted in about 50% higher fresh weights, and also increased the number of calli producing shoots by about 35%. More calli produced shoots in white light than in blue or red light or in darkness. The length of the photoperiod or intensity of light was not critical for shoot production.
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  • 35
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    Journal of Cellular Biochemistry 41 (1989), S. 179-188 
    ISSN: 0730-2312
    Keywords: protein kinase C ; TPA ; cell transformation ; 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 used a previously described retroviral expression vector pMV7-PKCβ1 to develop derivatives of two rat liver epithelial cell lines, K16 and K22, that stably express about tenfold-higher PKC activity than control cells. Despite these high levels of PKC, these cells did not exhibit gross morphologic changes, anchorage-independent growth, or tumorigenicity. K16PKC-4 and K22PKC-2, two lines with the highest PKC enzyme activity, were studied further in terms of several responses to the phorbol ester tumor promoter TPA. When treated with 100 ng/ml of TPA, the control K16MV7 and K22MV7 cells displayed a slight change in morphology, whereas the K16PKC-4 and K22PKC-2 cells displayed a marked change in morphology. Northern blot analyses demonstrated that TPA induced increased levels of fos, myc, phorbin, and ODC RNAs in control K16MV7 and K22MV7 cells, with maximum induction occurring at about 0.5, 1, 8, and 8 h, respectively. In K16PKC-4 and K22PKC-2 cells, TPA induction of phorbin and ODC RNAs was markedly enhanced, but this was not the case for myc and fos RNAs. In addition, the levels of myc RNA were constitutively higher in both K16PKC-4 and K22PKC-2 cells than in the control cells. Taken together, these results provide direct evidence that PKC plays a critical role in modulating the expression of myc, phorbin, and ODC RNAs. On the other hand, overexpression of PKCβ1 is not itself sufficient to cause cell transformation.
    Additional Material: 3 Ill.
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  • 36
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    Cellular and molecular life sciences 44 (1988), S. 910-919 
    ISSN: 1420-9071
    Keywords: Heart embryology ; developmental biology ; differentiation ; morphogenesis ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The subject of heart development has attracted the interest of many embryologists over the last two centuries. As a result, the main morphologic features of the developmental anatomy of the heart are already well established. Although there are still some controversial points, and there is probably much descriptive work yet to be done, emphasis is currently being placed on developmental mechanisms rather than simply on descriptive facts. The availability of new techniques and the overall advances in biological research are placing heart embryology in a new perspective. Today, we do not simply ask whether one or another embryonic structure arises further right or further left; instead, we are studying how cells, tissues, and their microenvironment interrelate at the several levels of biological organization (from the gene upwards) so as to give rise to a mature organ with a distinct shape and well-established functions. This paper attempts to review some of the basic aspects of the developmental anatomy of the heart. Descriptive embryology is used here as a tool. Emphasis is placed on developmental mechanisms, and on the present knowledge of how these mechanisms are related to the structural development of the heart.
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  • 37
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    Cellular and molecular life sciences 44 (1988), S. 930-936 
    ISSN: 1420-9071
    Keywords: Cardiac development ; embryogenesis ; gene expression ; complementary DNA ; molecular methodologies ; myocardial contractility ; myosin ; atrial natriuretic factor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Cardiac development involves a complex integration of subcellular processes into multicellular and, finally, whole organ effects. Until recently it has been difficult to investigate the genetic control of this organ level differentiation of the heart. The proliferation of molecular biology methodologies has provided mechanisms to directly investigate the control of these processes. This article focuses on molecular lines of research on two key areas in cardiac development: the regulation of expression of sarcomeric contractile and regulatory proteins, and atrial natriuretic factor. Molecular approaches are described which have allowed investigators to begin to determine the tissue and stage-specific expression of genes, to locate those genes in the genome, determine their sequences, and to directly investigate the mechanisms controlling their expression.
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  • 38
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    Cellular and molecular life sciences 44 (1988), S. 348-351 
    ISSN: 1420-9071
    Keywords: L. lactis ; protoplasts ; protoplast regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary More than 99% ofL. lactis cells have been converted to protoplasts upon digestion of cell walls with mutanolysin (N-(acetyl)muramidase). Functional protoplasts were obtained even with the lowest level of the enzyme that was used (0.1 U·ml−1 of the cell suspension) and after incubation at 37°C for 2 min. The regeneration of the polymerized cell wall appears to be induced by a cell homogenate of the same organism.
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  • 39
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    Cellular and molecular life sciences 44 (1988), S. 52-55 
    ISSN: 1420-9071
    Keywords: Human sperm ; human protamine ; protamine 1 ; protamine 2 ; protamine 3 ; infertility ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Protamines were extracted from the sperm of fertile and infertile human males and the relative proportion of protamines 1, 2, and 3 were determined by scanning microdensitometry following electrophoresis of total protamine in polyacrylamide gels. The proportion of the three protamines was found to be similar in sperm obtained from different normal males. The distribution of protamines in sperm obtained from a select group of infertile males producing an elevated level of large sperm heads, in contrast, was different from that of the fertile males.
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  • 40
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    Molecular and cellular biochemistry 84 (1988), S. 3-15 
    ISSN: 1573-4919
    Keywords: prostate ; androgens ; gene expression ; in situ hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary After castration, the rat dorsolateral prostate M-40 mRNA initially decreased then rebounded to precastrated levels. The cellular site of M-40 expression and its renewed expression after castration was defined by in situ hybridization histochemistry. In situ hybridization with either a 32P-labeled or biotin-labeled M-40 cDNA probe demonstrated that M-40 mRNA levels were higher in the lateral than dorsal prostate. A second androgen regulated gene, RWB, also was highly expressed in the lateral prostate. The biotinylated cDNA probes provided microscopic resolution of the expressing cells, revealing two distinct morphologies of lateral epithelium which expressed both the M-40 and RWB mRNA. These morphologies appeared in ducts which contained either epithelial cell sheets that were highly convoluted or thinner epithelial cells with a minimal degree of convolution. The RWB mRNA decreased in both cell populations in response to androgen withdrawal. The decline and reappearance of M-40 mRNA also appeared in both epithelial cell types. These data demonstrated that after castration the M-40 mRNA initially decreased as expected for an androgen sensitive gene and then progressed to a fully inducible state. The mechanism of this progression remains to be elucidated.
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  • 41
    ISSN: 1573-4927
    Keywords: cat hemoglobins ; polymorphism ; DNA ; restriction endonuclease ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract The molecular basis for the genetic control of variable proportions of the two cat hemoglobins Hb A(α2β 2 A ) and Hb B (α2β 2 B ) was investigated. Ratios of Hb A/Hb B vary between 50/50 and 90/10 among members of the mongrel cat population, with clusters around 50, 35, and 10% Hb B. Genomic DNA from cats of 50/50, 70/30, and 90/10 phenotypes were cut by restriction endonucleasesHindIII,EcoRI,BamHI,Bg1II,and Pst1 and hybridized to a fragment of the human β-globin gene. The results of the Southern blots suggested a pattern of homozygote, heterozygote, homozygote for the respective cat phenotypes, 50/50, 70/30, and 90/10. Therefore, the cat hemoglobin polymorphism seems to result from the possible combinations of an allelic gene pair.
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  • 42
    ISSN: 1573-5028
    Keywords: barley ; chymotrypsin inhibitor ; gene expression ; endosperm
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Full-length cDNA clones for barley chymotrypsin inhibitor 1 (CI-1) have been isolated from an endosperm-specific cDNA library. Hybridization and nucleotide sequence analyses indicate that these cDNAs represent two distinct types of CI-1 mRNA which we have called CI-1A and CI-1B. Both mRNAs encode polypeptides of 83 residues (M r=8790 and 8960) which differ at eleven positions. The full-length cDNA sequences do not predict N-terminal signal peptide extensions indicating that CI-1 is synthesized in the mature form in contrast to the homologous proteinase inhibitors of tomato and potato. Northern hybridization experiments show that the CI-1 genes are under strict developmental and organ-specific control. CI-1 transcripts were first detected in the developing barley endosperm between 12 and 14 days after anthesis but no CI-1-related sequences were detected in the RNA preparations from shoots, leaves or roots. The expression of CI-1 was also studied in the high-lysine barley mutants Hiproly, Risø 56 and Risø 1508. Approximately 15-fold (Hiproly) and 4-fold (Risø 56 and 1508) higher levels of CI-1 mRNA were detected in the mutant endosperms compared to normal barley. These results correlate well with the increased deposition of CI-1 in the high-lysine lines and show that the differential expression is controlled mainly at the level of transcription or stability of the mRNA. Using Southern-blots of barley DNA we estimate that there are three copies of CI-1 per haploid genome in both normal and mutant barley lines.
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  • 43
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    Plant molecular biology 11 (1988), S. 641-649 
    ISSN: 1573-5028
    Keywords: codon usage ; gene expression ; histone H3 gene ; Medicago sativa ; somatic embryos
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A histone H3 gene was isolated from a dicotyledonous plant, alfalfa (Medicago sativa). The sequence analysis of this gene revealed no obvious GC preference in its codon usage. Apart from containing most of the typical consensus sequences found in both animal and plant histone genes, the alfalfa H3 gene exhibits distinct structural features such as (1) the unusual location of two GATCC motifs in its 5′ flanking sequence, (2) the existence of a CGCGGATC on the nonsense strand at position −232, (3) the existence of a long palindromic structure, and (4) several polyadenylation signal-like sequences in the 3′ flanking region. There are about 160 copies of histone H3 gene in alfalfa tetraploid genome. Using the alfalfa H3 gene as a probe to study the pattern of histone H3 transcripts in the alfalfa, we found that the H3 RNAs are undetectable in leaves, more in stems than in roots, and highest in somatic embryos. Moreover, the RNA products of H3 genes in all alfalfa tissues tested show unusually long nontranslated region compared to those of animal histone genes. An additional high molecular weight species of H3 transcript was detected only in somatic embryos.
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  • 44
    ISSN: 1573-5028
    Keywords: Agrobacterium ; gene expression ; legumin (Pisum) ; Nicotiana ; seed storage protein ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A 3.4-kilobase genomic DNA fragment from Pisum sativum L. containing the LegA gene, which encodes a major legumin storage protein, was transferred to Nicotiana plumbaginifolia using an Agrobacterium tumefaciens strain containing the Bin 19 binary vector system. Northern hybridisation analysis of legA-transformed plants demonstrated that legumin-specific RNA was present in developing seeds but not in developing leaves. Legumin protein was immunologically detected in the mature seeds of legA-transformed plants, and was present as the correct-size protein composed of disulphide-bonded polypeptides. It is concluded that the transferred pea genomic fragment contains all the information necessary for seed-specific expression of the legA gene, and for correct processing of the primary transcript and the precursor legumin protein.
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  • 45
    ISSN: 1573-5028
    Keywords: potato ; patatin ; proteinase inhibitor ; cloning ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Tuberization in potato is a complex developmental process involving the expression of a specific set of genes leading to the synthesis of tuber proteins. We here report the cloning and analysis of mRNAs encoding tuber proteins. From a potato tuber cDNA library four different recombinants were isolated which hybridized predominantly with tuber mRNAs. Northern blot hybridization experiments showed that three of them, pPATB2, p303 and p340, can be regarded as tuber-specific while the fourth, p322, hybridizes to tuber and stem mRNA. Hybrid-selected in vitro translation and nucleotide sequence analysis indicate that pPATB2 and p303 represent patatin and the proteinase inhibitor II mRNA respectively. Recombinant p322 represents an mRNA encoding a polypeptide having homology with the soybean Bowman-Birk proteinase inhibitor while p340 represents an mRNA encoding a polypeptide showing homology with the winged bean Kunitz trypsin inhibitor. In total, these four polypeptides constitute approximately 50% of the soluble tuber protein. Using Southern blot analysis of potato DNA we estimate that these mRNAs are encoded by small multigene families.
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  • 46
    ISSN: 1573-5028
    Keywords: cDNA sequence ; chloroplast protein import ; gene expression ; glutamine synthetase ; nitrogen metabolism ; Phaseolus vulgaris L.
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Work using a full-length cDNA clone has revealed that the plastid-located glutamine synthetase (GS) of Phaseolus vulgaris is encoded by a single nuclear gene. Nucleotide sequencing has shown that this cDNA is more closely related to a cDNA encoding the plastidic GS of Pisum sativum than to cDNAs encoding three different cytosolic GS subunits of P. vulgaris. The plastid GS subunits are initially synthesized as higher M r (47000) precursors containing an N-terminal presequence of about 50 amino acids which is structurally similar to the presequences of other nuclear-encoded chloroplast proteins. The precursor has been synthesized in vitro and is imported by isolated pea chloroplasts and processed to two polypeptides of the same size as native P. vulgaris chloroplast GS subunits (M r 42000). Experiments with fusion proteins show that the N-terminal 68 amino acids of this precursor allow the cytosolic GS subunit β also to be imported and processed by isolated chloroplasts. Polyadenylated mRNA specifically related to the plastidic GS gene is most highly abundant in chloroplast-containing organs (leaves and stems) but is also detectable in roots and nodules.
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  • 47
    ISSN: 1573-5028
    Keywords: glyphosate ; herbicide ; 5-enolpyruvylshikimic acid 3-phosphate (EPSP) synthase ; enzyme overproduction ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell cultures of Corydalis sempervirens adapted to growth in the presence of 5 mM glyphosate [N-(phosphonomethyl)glycine] display a 30- to 40-fold increase in the cellular content of 5-enolpyruvylshikimic acid 3-phosphate (EPSP) synthase, the target enzyme of the herbicide. Translatable mRNA activity as well as transcript levels for EPSP synthase were increased 8-to 12-fold in the adapted (glyphosate-tolerant) as compared to the non-adapted (glyphosate-sensitive) cultures. Northern blot analysis revealed a single 1.8 kb transcript after hybridization with an oligonucleotide probe deduced from the N-terminal amino acid sequence of the enzyme. No significant differences in the relative abundance of EPSP synthase-specific DNA sequences could be detected, however, in Southern and dot blot analyses of restricted DNA isolated from the two cultures. We conclude that the overproduction of EPSP synthase in glyphosate-tolerant C. sempervirens cells is not based on the amplification of the corresponding gene.
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  • 48
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    Plant molecular biology 11 (1988), S. 495-506 
    ISSN: 1573-5028
    Keywords: abscisic acid ; barley aleurone layers ; cDNA cloning ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Abscisic acid (ABA) inhibits the gibberellic acid induced synthesis of α-amylase in barley aleurone layers, yet ABA itself induces more than a dozen polypeptides (Lin & Ho, Plant Physiol 82: 289–297, 1986). As part of our effort to elucidate the molecular action of ABA in barley aleurone layers, we have isolated and characterized an ABA-induced cDNA clone, pHV A1. This cDNA clone hybridizes to an RNA species of approximately 1.1 kb from ABA-treated barley aleurone layers. The level of this mRNA is tripled within 40 minutes after ABA treatment, reaches a peak at 8–12 h, and is present up to 48 h. The induction of this mRNA responds to concentrations of ABA as low as 10-9 M, but higher ABA concentrations induce higher expression of this mRNA. The products of hybrid-select translation and in vitro transcription/translation with pHV A1 comigrate on SDS gel as a 27 kDa polypeptide. However, the sequence of pHV A1 indicates that it has an open reading frame encoding a 22 kDa protein. This size discrepancy is probably due to the high content of the basic amino acid, lysine. This notion has been confirmed by two-dimensional gel electrophoresis showing that this polypeptide is one of the most basic proteins in ABA-treated barley aleurone layers. The deduced amino acid sequence of pHV A1 contains nine imperfect repeats 11 amino acids long which share homology with cotton Lea 7 protein (Baker, Steele & Dure, Plant Mol Biol, in press). The identity and function of the encoded product of pHV A1 is under investigation.
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  • 49
    ISSN: 1573-0778
    Keywords: CG suppression ; DNA methylation ; gene expression ; MHC ; mutation hot spots
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract DNA methylation of human class II genes of the Major Histocompatibility Complex (MHC) was correlated with gene expression and methyl-related CG → TG and CG → CA changes. It was found that cytosine methylation of the CG dinucleotides of MHC class II genes should be involved in generating a fraction of nucleotide polymorphism, rather than in controlling transcription.
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  • 50
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    Photosynthesis research 18 (1988), S. 7-31 
    ISSN: 1573-5079
    Keywords: Nicotiana ; chloroplast genome ; photosynthesis ; NADH dehydrogenase ; protein structure ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In order to understand more fully chloroplast genetic systems, we have determined the complete nucleotide sequence (155, 844 bp) of tobacco (Nicotiana tabacum var. Bright Yellow 4) chloroplast DNA. It contains two copies of an identical 25,339 bp inverted repeat, which are separated by 86, 684 bp and 18,482 bp single-copy regions. The genes for 4 different rRNAs, 30 different tRNAs, 44 different proteins and 9 other predicted protein-coding genes have been located. Fifteen different genes contain introns. Twenty-two genes for components of the photosynthetic apparatus have so far been identified. Most of the genes (except the gene for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase) code for thylakoid membrane proteins. Twenty of them are located in the large single-copy region and one gene for a 9-kd polypeptide of photosystem I is located in the small single-copy region. The gene for the 32-kd protein of photosystem II as well as the gene for the large subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase have strong promoters and are transcribed monocistronically while the other genes are transcribed polycistronically. We have found that the predicted amino acid sequences of six DNA sequences resemble those of components of the respiratory-chain NADH dehydrogenase from human mitochondria. As these six sequences are highly transcribed in tobacco chloroplasts, they are probably genes for components of a chloroplast NADH dehydrogenase. These observations suggest the existence of a respiratory-chain in the chloroplast of higher plants.
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  • 51
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    Photosynthesis research 19 (1988), S. 63-71 
    ISSN: 1573-5079
    Keywords: CO2 fixation ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract From studies conducted in both our laboratory and by Gibson, Tabita and colleagues, as well as drawing on the recent studies with Alcaligenes eutrophus, we describe two genetic regions which have been identified on the chromosome of Rhodobacter sphaeroides which code for a number of enzymes involved in CO2 fixation. One region was found to contain the genes coding for fructose 1,6-bisphosphatase (fbpA), phosphoribulokinase (prkA), a 37 kDa polypeptide (cfxA), and form I ribulose 1,5-bisphosphate carboxylase/oxygenase (rbcL, S). These genes appear to be expressed in the same transcriptional direction and are tandomly arranged. A second, apparently unlinked region of the chromosome contains a duplicate (with respect to functionality of gene products) but not identical set of these same four genes. Although the gene order in both regions is apparently identical, there is approximately 4 kb of DNA separating the 3′-end of prkB and the beginning of cfxB. The specific genetic organizations and proposed roles of these two genetic regions are discussed.
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  • 52
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    Photosynthesis research 19 (1988), S. 129-152 
    ISSN: 1573-5079
    Keywords: chloroplast biogenesis ; gene expression ; ribosomes ; transcription ; translation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Isolated chloroplasts show substantial rates of protein synthesis when illuminated. This ‘in organello’ protein synthesis system has been advantageously utilised to elucidate the coding capacity of chloroplast and the regulation of chloroplast genes. The system is also being used recently to transcribe and translate homologous and heterologous templates. In this mini-review, we attempt to critically ecaluate the available literature and present the current and the prospective lines of research.
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  • 53
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    Photosynthesis research 18 (1988), S. 205-222 
    ISSN: 1573-5079
    Keywords: ATP synthase ; chloroplast ; evolution ; gene expression ; operon
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three CF1 and three CF0 subunits of the chloroplast H+-ATP synthase are encoded on the chloroplast genome. The chloroplast atp genes are organized as two operons in plants but not in the green alga, Chlamydomonas reinhardtii. The atpBE or β operon shows a relatively simple organisation and transcription pattern, while the atpIHFA or α operon is transcribed into a large variety of mRNAs. The atp genes are related to those of cyanobacteria and, more distantly, to those of non-photosynthetic bacteria such as E. coli, suggesting a common origin of most F1F0 ATP synthase subunits. Both the chloroplast and cyanobacterial ATP synthases have four F0 subunits, not three as in the E. coli complex. The proton pore of the CF0 is proposed to be formed by the interaction of subunits III and IV.
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  • 54
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    Photosynthesis research 17 (1988), S. 145-157 
    ISSN: 1573-5079
    Keywords: Carbon fixation ; enzyme assembly ; gene expression ; recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The synthesis in Escherichia coli of both the large and small subunits of cereal ribulose bisphosphate carboxylase/oxygenase has been obtained using expression plasmids and bacteriophages. The level and order of synthesis of the large and small subunits were regulated using different promoters, resulting in different subunit pool sizes and ratios that could be controlled in attempts to optimize the conditions for assembly. Neither assembly nor enzyme activity were observed for the higher plant enzyme. In contrast, cyanobacterial large and small subunits can assemble to give an active holoenzyme in Escherichia coli. By the use of deletion plasmids, followed by infection with appropriate phages, it can be demonstrated that the small subunit is essential for catalysis. However, the small subunit is not required for the assembly of a large subunit octomer core in the case of the Synechococcus enzyme; self-assembly of the octomer will occur in an rbcS deletion strain. The cyanobacterial small subunits can be replaced by wheat small subunits to give an active enzyme in Escherichia coli. The hybrid cyanobacterial large/wheat small subunit enzyme has only about 10% of the level of activity of the wild-type enzyme, reflecting the incomplete saturation of the small subunit binding sites on the large subunit octomer, and possibly a mismatch in the subunit interactions of those small subunits that do bind, giving rise to a lower rate of turnover at the active sites.
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  • 55
    ISSN: 1573-5044
    Keywords: Brassica napus ; protoplasts ; Ficoll ; agarose
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A method is described for regenerating callus from mesophyll protoplasts of a winter variety of Brassica napus. The method combines the use of Ficoll in an initial liquid medium, enhancing early protoplast division and cell colony formation, with a transfer to an agarose system after 10 days culture to give rapid microcalli formation. Further transfers resulted in callus regeneration and the initiation of organogenesis.
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  • 56
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    Plant cell, tissue and organ culture 14 (1988), S. 15-24 
    ISSN: 1573-5044
    Keywords: Brassica ; genetics of regeneration ; protoplasts ; totipotency
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Factors affecting the division of cells derived from leaf and cotyledon protoplasts from Brassica oleracea L. var. italica (‘Green Comet’ hybrid broccoli) were examined to optimize conditions for plant regeneration and to determine whether there was a genetic basis for improved regeneration from protoplasts derived from plants previously regenerated from tissue cultures [15]. When leaf protoplasts from different plants grown from hybrid seed were isolated and cultured simultaneously, division efficiencies of 1–95% were obtained. Cells from some plants showed high division efficiencies in consecutive experiments while cells from other plants had consistently low division rates. More plants from hybrid seed gave high division efficiencies when cotyledon protoplasts were used. However, cotyledon or leaf protoplasts from selfed progeny of regenerated plants produced more vigorous calli and more shoots than protoplasts from hybrid seed. These results suggest that there may be a genetic component to the increased totipotency of Brassica oleracea protoplasts.
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  • 57
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    Journal of bioenergetics and biomembranes 20 (1988), S. 19-39 
    ISSN: 1573-6881
    Keywords: E. coli unc operon ; H+-ATPase ; subunit stoichiometry ; gene expression ; codon usage ; translational initiation ; Shine-Dalgarno sequence ; recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Physics
    Notes: Abstract Theunc (or atp) operon ofEscherichia coli comprises eight genes encoding the known subunits of the proton-translocating ATP synthase (H+-ATPase) plus a ninth gene (uncI) of unknown function. The subunit stoichiometry of the H+-ATPase (α 3β3γ1δ1ε1a1b2c10–15) requires that the respectiveunc genes be expressed at different rates. This review discusses the experimental methods applied to determining how differential synthesis is achieved, and evaluates the results obtained. It has been found that the primary level of control is translational initiation. The translational efficiencies of theunc genes are determined by primary and secondary mRNA structures within their respective translational initiation regions. The respective rates of translation are matched to the subunit requirements of H+-ATPase assembly. Finally, points of uncertainty remain and experimental strategies which will be important in future work are discussed.
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  • 58
    ISSN: 1573-7241
    Keywords: Atrial natriuretic peptide ; cardiac myocytes ; gene expression ; hybridization
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Primary cultures of neonatal cardiocytes expreses the gene for atrial natriuretic peptide (ANP). In general the levels of expression follow the rank order: atrial 〉 ventricular ≫ nonmyocardial cells. Following the initial dispersion cardlocytes require 48 to 72 hours before ANP seeretion and ANP mRNA accumulation approach a new steadystate level. In situ hybridization analysis indicates that ANP gene expression is concentrated in a subpopulation of cardiocytes in both the atrial and the ventricular cell cultures. These findings suggest that these primary cultures may be of value in defining the faciors governing the expression of the ANP gene in the cardiac cell.
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  • 59
    ISSN: 1573-5079
    Keywords: Crassulacean acid metabolism ; gene expression ; Mesembryanthemum crystallinum ; mRNA levels ; soil salinity
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mesembryanthemum crystallinum responds to high salinity in the soil by shifting the mode of carbon assimilation from the C3 mode to Crassulacean acid metabolism (CAM). Several enzymes of carbon metabolism have increased apparent activities in the CAM mode, including phosphoenolpyruvate carboxylase (PEPcase) and pyruvate orthophosphate dikinase (PPDK). We have identified cDNA clones for PEPcase and PPDK by immunological screening of a cDNA library constructed in the protein expression vector lambda gt11. The clones were characterized by immunoblotting and RNA blotting techniques. RNA blotting showed that during CAM induction the steady-state level of mRNAs for both PEP case and PPDK increased.
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  • 60
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    Photosynthesis research 19 (1988), S. 39-61 
    ISSN: 1573-5079
    Keywords: Bacteriochlorophyll-protein complex ; gene expression ; light-harvesting complex ; reaction center ; Rhodospirillaceae ; transcriptional control ; translational control
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The puf operon of the purple nonsulfur photosynthetic bacterium, Rhodobacter sphaeroides, contains structural gene information for at least two functionally distinct bacteriochlorophyll-protein complexes (light harvesting and reaction center) which are present in a fixed ratio within the photosynthetic intracytoplasmic membrane. Two proximal genes (pufBA) specify subunits of a long wavelength absorbing (i.e., 875 nm) light harvesting complex which are present in the photosynthetic membrane in ≃15 fold excess relative to the reaction center subunits which are encoded by the pufLM genes. This review summarizes recent studies aimed at determining how expression of the R. sphaeroides puf operon region relates to the ratio of individual bacteriochlorophyll-protein complexes found within the photosynthetic membrane. These experiments indicate that puf operon expression may be regulated at the transcriptional, post-transcriptional, translation and post-translational levels. In addition, this review discusses the possible role(s) of newly identified loci upstream of pufB which may be involved in regulating either synthesis or assembly of individual bacteriochrlorophyll-protein complexes as well as the pufX gene, the most distal genetic element within the puf operon whose function is still unknown.
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    Journal of Cellular Biochemistry 37 (1988), S. 243-253 
    ISSN: 0730-2312
    Keywords: hepatocyte differentiation ; gene expression ; hormonal control ; glucocorticoids ; insulin ; cyclic AMP ; tyrosine aminotransferase ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Cloned cDNAs were used in hybridization analyses to assess hormonal responsiveness of two similarly regulated genes in livers of late-term fetal rats. Transcription of the tyrosine aminotransferase gene and of gene 33 (Lee et al.: J Biol Chem 260:16433-16438, 1985) is enhanced by glucocorticoids and by each of the usually antagonistic hormonal agents, insulin and cAMP, in adult liver, and that of both genes is developmentally activated at or just prior to birth. The mRNA of gene 33 was found to be significantly increased by each of the hormonal regulators in livers of fetuses treated in utero. Expression of the nearly silent aminotransferase gene in fetal liver was appreciably increased by cAMP but was refractory to control by either glucocorticoids or insulin; capacity of this gene to respond to insulin was not realized until several days postpartum. The data indicate specificity in the developmental acquisition of the capacity of individual genes to respond to hormonal regulators.
    Additional Material: 9 Ill.
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  • 62
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    Journal of Cellular Biochemistry 38 (1988), S. 205-212 
    ISSN: 0730-2312
    Keywords: cytokines ; gene expression ; retroviral promoters ; antitumoral activities ; phenotypical reversion ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The potential mechanisms of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha action on tumor cells have been investigated in a model of mouse fibroblasts transformed by distinct retroviral vectors carrying the v-mos, c-myc, and v-Ha-ras oncogene, respectively. Treatment with both cytokines not only caused growth inhibition of v-mos- and c-myc-transformants, but also a reversion of transformation-induced suppression of major histocompatibility complex (MHC) class I antigen expression in all transformed cell lines. The phenotypical reversion of transformants was preceded by a selective modulation of LTR-controlled oncogene expression. TNF-alpha primarily affected stability of oncogene-specific RNAs without influencing the activity of retroviral promoters. In contrast, IFN-gamma was effective at the transcriptional level, apparently due to inhibition of LTR activity as revealed from reduced CAT activity in IFN-gamma-treated LTR-CAT transformants. This IFN-gamma-mediated down-regulation of retroviral promoter activity seemed to be selective for Moloney-virus-derived promoters, since the activity of other viral and cellular promoters was not suppressed by IFN-gamma.
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  • 63
    ISSN: 0730-2312
    Keywords: ADP-ribosyl transferase ; cell proliferation ; gene expression ; protein kinase C ; c-myc expression ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Human α or β interferons inhibit the proliferation of Daudi Burkitt lymphoma cells and induce the differentiation of these cells towards a mature plasma cell phenotype. Similar responses are seen when Daudi cells are treated with the phorbol ester, TPA. Both interferons and TPA down-regulate expression of the c-myc oncogene in these cells. Although TPA can mimic the effect of interferon on cell differentiation, it does not induce 2′5′ oligoadenylate synthetase or the interferon-sensitive mRNAs, 6-16 or 9-27. Thus chronic stimulation of protein kinase C by TPA cannot mimic all of the effects of interferon treatment on gene expression. Inhibition of ADP-ribosyl transferase activity by 3-methoxybenzamide impairs interferon- or TPA-induced differentiation of Daudi cells. This agent induces a higher level of c-myc mRNA in the cells and stimulates the incorporation of [3H]thymidine into DNA; although these effects are partially counteracted by interferon or TPA treatment, the elevated expression of the c-myc gene may be sufficient to prevent terminal differentiation and allow cell proliferation to continue.
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    Developmental Genetics 9 (1988), S. 13-22 
    ISSN: 0192-253X
    Keywords: ontogeny ; lethality ; gene expression ; mRNA ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cadmium is a toxic metal that induces the expression of metallothionein genes in many tissues and that binds avidly to metallothionein, a soluble transition metal binding protein. The present study examined the temporal pattern and magnitude of accumulation of metallothionein mRNA in liver of C57BL/6J mice of various ages treated with cadmium. In adult female mice, accumulation was dependent on the dosage level of cadmium and related to the concentration of this metal in liver. The accumulation of metallothionein mRNA in liver depended on age at exposure to cadmium. Intraperitoneal administration of 2 mg of cadmium per kg provoked small increases (two- to threefold) in levels of metallothionein mRNA in livers of 7- and 14-day-old mice. In contrast, cadmium treatment of 28- and 56-day-old mice resulted in 12- to 19-fold increases in levels of metallothionein mRNA in liver with maximum increases occurring 3 to 4 hr after treatment. Because similar patterns for the accumulation of cadmium of liver were found in 7-, 28-, and 56-day-old mice, observed age-dependent differences in induction of metallothionein mRNA in liver were probably not due to differences in the accumulation of cadmium in this organ. Taken together, these data suggest that tissue-specific factors controlling the expression of metallothionein genes may account for developmental variation in the inducibility of these genes by cadmium. Ontogenic variation in accumulation of metallothionein mRNA after cadmium treatment may be a factor in developmental variation in the acute lethality of cadmium in C57BL/6J mice.
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  • 65
    ISSN: 0192-253X
    Keywords: Daucus carota ; auxin ; gene expression ; mutant ; filtration-enrichment procedure ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Cell cultures of the carrot Daucus carota are a useful experimental system for studying the genetic regulation of plant embryogenesis. A modified filtration-enrichment procedure was used to isolate 21 temperature-sensitive variants in somatic embryogenesis; the variants display normal embryo development at the permissive temperature (24°C) and altered development at the restrictive temperature (33°C). Temperature-shift experiments were performed on these variants to determine the timing of gene action for the putative temperature-sensitive alleles. According to their phenotypes at the restrictive temperature, these variants can be divided into six classes: No Growth, Callus Proliferation, Globularstage Block, Oblong-stage Block, Lateral Growth, and Root Formation. Although many variants exhibit lengthy temperature-sensitive periods, the temperature sensitivity of some variants is restricted to one or two embryonic stages. These results plus those in the literature are incorporated into a preliminary model concerning the genetic regulation of carrot embryogenesis.
    Additional Material: 9 Ill.
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    Developmental Genetics 9 (1988), S. 259-265 
    ISSN: 0192-253X
    Keywords: G-proteins ; gene expression ; developmental regulation ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: We have identified a two-member gene family in the Dictyostelium genome and have isolated corresponding cDNA or genomic DNA recombinant clones. Analyses of these DNA sequences predicted encoded proteins of ∼200 amino acids with ∼90% sequence identity to each other. These Dictyostelium proteins also share amino acids identity within the GTP-binding domains in the family of G-regulatory proteins involved in cellular regulation and transmembrane signalling. Additional structural similarities are seen with members of the ras supergene family, such as ras, ral, and rho. They are similar in size (usually ∼200 amino acids), possess four conserved domains involved in GTP interaction and are believed to be anchored in the membrane by fatty acid modification of a cysteine residue near the carboxy terminus. More extensive identity is observed with YPT1 and SEC4, two other members of this family of genes that are essential in yeast. The amino-terminal half of both Dictyostelium proteins is 70% identical in amino acid sequence to the YPT1 and SEC4 yeast proteins with less identity continuing through the remainder of the proteins. In addition these proteins terminate in two cysteine residues that are thought to be required for membrane anchorage.The two genes within this Dictyostelium family are organized differently in the genome and are differentially regulated during development. One gene is colinear in sequence with its mRNA in the protein coding region, whereas the other gene encodes a spliced mRNA. The intron-containing gene is associated with a developmentally regulated (AAC)-repeat sequence. Finally, we have shown that the expression of one of the genes is induced during development with kinetics similar to that of other (AAC)n-associated genes; conversely, the expression of the second gene is repressed at a similar developmental stage.
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  • 67
    ISSN: 0192-253X
    Keywords: cAMP ; Ca2+ ; signal transduction ; cell surface receptor ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Elevated levels of cAMP are essential for the expression of many postaggregation prespore and prestalk mRNA species and for the suppression of some growth phase mRNAs. Here we review evidence that this regulation is mediated by cAMP interacting at the cell surface receptor. These effects of cAMP on gene expression can occur under conditions where the receptor-associated adenylate cyclase is inactivated and in concentrations that are consistent with receptor binding. A number of differences are noted in the mechanism by which cAMP regulates prespore and prestalk genes. Finally, evidence is reviewed for the role of a Ca2+-dependent signal transduction system in coupling the expression of some of the prespore mRNAs to the cAMP receptor. This signal transduction system does not appear to be involved in the expression of the cAMP-dependent prestalk gene.
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    Developmental Genetics 9 (1988), S. 167-180 
    ISSN: 0192-253X
    Keywords: enzyme pattern ; gene expression ; protein synthesis ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Aldehyde oxidase (AO) and pyridoxal oxidase (PO) distribution patterns were determined in the imaginal wing discs for a series of strains of Drosophila melanogaster heterozygous for different Minute mutations. The mutant severity ranged from very weak to strong. The results show an inverse response of AO and PO to the expressivity of the Minute mutation: in weaker Minutes the extent of the AO positive area increases, whereas PO activity disappears. The results are discussed with reference to an impaired protein synthesis in Minutes.
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    Cellular and molecular life sciences 43 (1987), S. 741-750 
    ISSN: 1420-9071
    Keywords: Hybridization histochemistry ; hybridocytochemistry ; in situ hybridization ; gene expression ; histochemical hybridization ; nucleic acid hybridization ; histocytochemistry
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The location of gene expression by hybridization histochemistry is being applied in many areas of research and diagnosis. The aim of this technique is to detect specific mRNA in cells and tissues by hybridization with a complementary DNA or RNA probe. Requirements for optimal specificity, sensitivity, resolution and speed of detection may not all be encompassed in one simple technique suitable for all applications, thus appropriate procedures should be selected for specific objectives. With reference to published procedures and our own extensive experience, we have evaluated fixatives, probes, labels and other aspects of the technique critical to the preservation and hybridization in situ of mRNA and detection and quantition of hybrids.
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    Molecular and cellular biochemistry 78 (1987), S. 3-8 
    ISSN: 1573-4919
    Keywords: polyamines ; RNA polymerase ; transcription ; gene expression ; mycobacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract All three polyamines — putrescine, spermidine and spermine stimulated the activity of mycobacterial RNA polymerase in vitro although the concentration required for maximal stimulation was different for each of the amines. Spermidine and spermine showed a biphasic effect on the enzyme activity. Stimulation of RNA synthesis by spermidine occurs only at higher DNA template/enzyme ratio. Spermidine stimulates RNA synthesis by acting on the elongation phase of RNA synthesis but it had no effect on initiation phase. Addition of mycobacterial RNA to the assay mixture resulted in the inhibition of RNA polymerase activity and this inhibition could be reversed by spermidine suggesting that spermidine stimulates transcription by binding to nascent RNA and thus destabilizing the short DNA-RNA hybrid region.
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    Molecular and cellular biochemistry 75 (1987), S. 61-70 
    ISSN: 1573-4919
    Keywords: gene expression ; development ; tissue-specific expression ; cell cycle-dependent genes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Abstract 4F1, 2A9 and 2F1 represent three of a number of cDNA sequences which have been identified because their cognate RNAs markedly increase when quiescent cells in culture are stimulated with serum. Studies using a variety of cell culture systems have shown that the expression of these genes is modulated by various growth factors and mitogens and thus such genes are considered to be ‘growth-associated.’ Thus far, little information has been obtained with these in vitro systems about the function of these genes. In an attempt to begin to elucidate the role of these genes (if any) in the physiology of the normal cell, we have analyzed the levels of 4F1, 2A9 and 2F1 transcripts in a variety of differentiated organs and tissues of adult and fetal rats. Our results show that each of these growth-associated genes exhibits its own unique pattern of expression, unrelated to the proliferative activity of the tissue. These data suggest that these genes most likely do have specific functions in normal tissue in addition to their role in the induction of DNA synthesis in quiescent cells in culture.
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  • 72
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    Bioscience reports 7 (1987), S. 955-963 
    ISSN: 1573-4935
    Keywords: gene expression ; GC box ; negative regulation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract A DNA control sequence T G GGGCGG AAT GGC , or the “GC” box, has been described in the promoter regions upstream of a number of eukaryotic genes transcribed by polymerase II (for review, see Dynan, W. S. and Tjian, R.,Nature 316:774, 1985). The “GC” box can occur in single or multiple copies and is the binding site for a protein factor, Spl, which activates initiation of transcription. We have observed in the rainbow trout protamine gene 3′ to the TATA box, three “GC” boxes spaced at 80 bp intervals. The first is 5′ to the cap site and possesses the ability to “silence” transcription from the protamine promoter in constructs linking this promoter to the bacterial chloramphenicol acetyl transferase (CAT) coding sequence following transfection to COS-1 cells. A model is proposed to account for the silencing of the protamine gene in all tissues except developing sperm cells.
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    Potato research 30 (1987), S. 371-380 
    ISSN: 1871-4528
    Keywords: dihaploids ; protoclones ; protoplasts ; Solanum tuberosum ; somaclonal variation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Description / Table of Contents: Zusammenfassung Die Verwendung dihaploider Kartoffelklone (2n=2x=24) in Züchtungsprogrammen, vor allem die Rückkehr zum tetraploiden Grad, wird sehr oft durch deren fehlende Fertilität verhindert. Dieses Problem dürfte am elegantesten durch somatische Fusion der Dihaploiden überwunden werden. Als Ergebnis einer solchen Prozedur würden sich, ausser der Paarung beider Ploidiestufen, die Addition qualitativer und quantitativer Merkmale ergeben. Grösstes Hindernis für die Anwendung der Protoplastenfusion ist das Fehlen eines brauchbaren Selektionssystems für die Separierung von homo- und heterokaryotischen Fusionsprodukten. In der hier beschriebenen Methode wurde der sehr kräftige Wuchs einiger Kalli für die Vorselektion einiger vermuteter Hybriden verwendet. Nach Anwendung der Polyethylenglykol-Fusions-methode (PEG, Tabelle 1) konnten von fünf selektierten dihaploiden Klonen (P1–P5) grosse Zahlen von Kalli und Pflanzen regeneriert werden, obwohl die PEG-Behandlung einen negativen Einfluss auf die Regenerationsrate hatte (Tabelle 2). Insgesamt wurden nach PEG-Behandlung 115 Kalli als vermutliche Hybriden, ihrer extremen Wuchsleistung entsprechend, selektiert. Tabelle 3 vergleicht die Ploidiestufen dieser selektierten Klone mit der von unbehandelten Elternklonen. Wegen der somaklonalen Variation wurden auch von nicht fusionierten Protoplasten viele tetraploide Klone gefunden. Ihre Zahl war allerdings signifikant kleiner, und unter den nicht PEG-behandelten Protoplasten waren immer einige diploide vorhanden. Die Tabellen 4 und 5 zeigen die Merkmale von 9 und 10 selektierten Klonen (Hy 1–10) in vitro und in vivo für Sprosslänge, Zahl der Nodien, Blattfläche, Blattform, Zahl der Wurzeln und allgemeiner Wachstumsleistung. In allen Fällen waren die gemessenen Parameter bei den selektierten Klonen signifikant grösser als bei den Kontrollen. Folglich kann das stärkere Wachstum der selektierten Klone nicht nur mit somaklonaler Variation erklärt werden. Es ist ein starkes Indiz für die Hybridnatur. Das Isoenzym-Muster der Esterasen unterstreicht diese Schlussfolgerung. Den Ergebnissen zufolge ist es möglich, somatische Hybriden anhand ihrer hybriden Vitalität vorzuselektieren. Dies sollte die Möglichkeit zur Entdeckung somatischer Hybriden in ausreichender Häufigkeit für praktische Züchtungsprogramme erhöhen.
    Abstract: Résumé L'utilisation de clônes dihaploïdes (2n=2x=24) dans les programmes d'hybridation, et particulièrement le retour au niveau tétraploïde, est entravée par leur manque de fertilité. Ce problème pourrait être maitrisé élégamment par la fusion somatique de dihaploïdes. D'un tel procédé résulterait de plus l'héritage de caractères qualitatifs et quantitatifs apportés par le doublement de ploïdie. Le principal obstacle pour utiliser la fusion de protoplastes est l'absence d'un système de sélection approprié pour la séparation des homo et hétérokaryotes produits par la fusion. Par la méthode décrite dans cet article la grande vigueur de croissance de quelques cals a été mise à profit pour la présélection des présumés hybrides. Après l'adaptation du procédé de fusion au polyéthylène-glycol (PEG, tableau 1) sur cinq clônes dihaploïdes sélectionnés (P1–P5) un grand nombre de cals et de plantes pourrait être régénéré, bien que le traitement PEG ait une influence négative sur le taux de régénération (tableau 2). Au total 115 cals étaient sélectionnés après le traitement comme de présumés hybrides, en raison de leur extrème vigueur. Le tableau 3 compare les niveaux de ploïdie de ces clônes sélectionnés avec ceux des parents non traités. La variation somatique de protoplastes non fusionnés est également trouvée dans de nombreaux clônes tétraploïdes, leur nombre est cependant significativement plus petit, et parmi les protoclônes régénérés de protoplastes non traités quelques diploïdes sont toujours présents. Les tableaux 4 et 5 montrent les caractéristiques de 9 et 10 clônes sélectionnés (Hy 1–10) in vitro et in vivo, respectivement pour la longueur de pouses, le nombre de noeuds, la surface et la forme des feuilles, le nombre de racines et la vigueur générale. Dans tous les cas, les paramètres mesurés ont des valeurs significativement plus élevées dans les clônes sélectionnés que dans les témoins. En conséquence, leur vigoureuse croissance ne peut être expliquée seulement par la variation somatique mais elle constitue une indication sur la vigueur hybride. L'analyse des estérases souligne cette conclusion (figure 1). Ces résultats montrent qu'il est possible de présélectionner des hybrides somatiques par leur vigueur, ce qui augmente les possibilités de les détecter en quantité suffisante dans les programmes d'hybridation.
    Notes: Summary Tetraploid potato plants were regenerated after polyethylene-glycol-induced protoplast fusion between dihaploids. Hybrid vigour of the regenerated calli was used for preselection of fusion products. Nearly all the selected vigorous clones possessed chromosome counts at the tetraploid level. Fusion products were compared to the parental material to auto-fused plants of and to three protoclones expressing different degrees of somaclonal variation. The selected clones, where grown in vitro in growth rooms and in pots in the glasshouse, showed increased vigour compared to their parents, to auto-fused and to 4x protoclones. Plants of clones from very vigorous calli, when assessed by height, the number of nodes per plant, leaf morphology and tuber production, showed hybrid vigour. The hypothesis that superior clones result from heterokaryons after protoplast fusion or that they arise from other in vitro events such as somaclonal variation is discussed.
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  • 74
    ISSN: 1573-5028
    Keywords: azacytidine ; DNA/RNA hybridization ; gene expression ; marker rescue ; nopaline-synthase ; plant tumor cells ; Ti-plasmid
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell wall regenerating protoplasts from soybean cells kept in suspension culture were cocultivated with bacteria which were derived from the nopaline strain C58 of Agrobacterium tumefaciens. When the bacteria carried an oncogenic Ti-plasmid, about 5% of the surviving protoplasts were able to form calli on hormone-free agar in contrast to controls, where bacteria without Ti-plasmid were applied, and where no calli were formed. After isolation of DNA from hormone-independently growing cells further evidence for transformation was obtained by hybridization to Ti-plasmid specific RNA and by rescue of a segment with a bacterial resistance gene which had been inserted before into the T-DNA. Transfer of T-DNA harboring a neomycin-resistance gene activated by the nos-promoter resulted in calli growing on kanamycin. Verification of segments located at the left and the right part of the T-DNA indicated the presence of its entire length in transformed soybean cells. Expression of T-DNA genes was measured by the assay of nopaline-synthase. Cells cultured on agar had a much higher level of nopaline-synthase than fast growing cells in suspension culture. Transferring them to agar or treatment with azacytidine strongly increased synthesis of nopaline-synthase indicating a reversible repression presumably via a methylation mechanism.
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  • 75
    ISSN: 1573-5028
    Keywords: cucumber ; gene expression ; hydroxypyruvate reductase ; light regulation ; peroxisomal enzymes ; serine:glyoxylate aminotransferase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The development of peroxisomal enzymes in cotyledons of cucumber seedlings is strongly dependent on light. In light-grown seedlings, activities of two peroxisomal enzymes, hydroxypyruvate reductase (HPR) and serine: glyoxylate aminotransferase (SGAT), were barely detectable until three days postimbibition, after which time both activities increased rapidly and linearly for at least three days. In the dark, the activities of these enzymes increased slightly over the same time period, but only to about 5% to 10% of 7-day light-induced levels. When 51/2-day dark-grown seedlings were transferred into white light, activities of HPR and SGAT began to increase after approximately 8 h. HPR protein was shown by an immunoprecipitation assay to increase concurrently with enzymatic activity in both light- and dark-grown cotyledons. Immunoblotting results suggested that the amounts of SGAT-A and SGAT-B, the two subunits of SGAT, also developed along with SGAT activity. The relative levels of translatable mRNAs encoding HPR, SGAT-A, and SGAT-B were also light-dependent, and increased with a developmental pattern similar to enzyme activity and protein levels in light- and dark-grown cotyledons. In 51/2-day dark-grown cotyledons that were transferred to the light, translatable mRNAs for SGAT-A and SGAT-B began to increase within 1 h of illumination and continued of increase rapidly and linearly for the next 24 h in the light to a new steady-state level that was 45 times that of dark controls. Translatable HPR mRNA exhibited a biphasic pattern of accumulation, with a three-fold increase during the first 6 h of illumination, followed by an additional six-fold increase between 8 and 24 h. The accumulation of translationally active mRNA for both enzymes preceded the accumulation of the corresponding protein and enzyme activity by about 8 h. Our data suggest that the rise in enzyme activity depends on an increase in translatable mRNA for these enzymes and is regulated at a pretranslational level, most likely involving transcription of new mRNA.
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  • 76
    ISSN: 1573-5044
    Keywords: Humulus lupulus ; micro-calli ; protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A cell-wall digestion medium has been devised to isolate protoplasts from suspension cultures of Humulus lupulus. Conditions have been developed for colony formation from protoplasts and the plating efficiency determined in three types of agar and by two culture methods. Viable calli were produced only when protoplasts embedded in Seaplaque agarose were incubated in a defined liquid medium. HPLC analysis showed that none of the isolated colonies accumulated α-acids.
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  • 77
    ISSN: 1573-5028
    Keywords: gene expression ; nitrogen fixation ; nodulins ; Phaseolus vulgaris ; Rhizobium ; root-nodule development
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The identification of some nodule-specific host proteins (nodulins) from common bean (Phaseolus vulgaris L.), a tropical ureide-transporting legume, is described. Particularly, the existence and developmental expression of several abundant nodule-specific transcripts of P. vulgaris are shown, including leghemoglobin, nodulespecific uricase and a group that in vitro translates into a cluster of about 30 kDa products. The expression pattern of nodulins in effective (Fix+) nodules compared to ineffective (Fix-) ones is also presented. The modified expression of main nodulins observed between these nodules indicates that different levels and/or factors associated with their regulation are involved. The intracellular infection by Rhizobium as a decisive step in the induction of some P. vulgaris nodulins is discussed.
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  • 78
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    Plant molecular biology 9 (1987), S. 547-563 
    ISSN: 1573-5028
    Keywords: acetate ; blue light ; Chlamydomonas reinhardtii ; chlorophyll a/b-binding protein gene ; gene expression ; LHCP gene
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In Chlamydomonas reinhardtii, the chlorophyll a/b-binding proteins of photosystem II are encoded in the nucleus by a small family of genes. We have studied the expression of one gene, which we call cabII-1, in a green-in-the-dark strain, which can synthesize chlorophyll in the dark or light, and in a yellow-in-the-dark mutant strain, which is able to make chlorophyll only in the light. In light/dark synchronized cultures of both strains, cabII-1 mRNA abundance increases during the first 6 h of a 12-h light phase, remains high for several hours, then declines. A variety of illumination conditions have been used to analyze the cabII-1 mRNA increase: continuous or intermittent red, blue, or white light, with or without 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU), an inhibitor of photosystem II. Our results suggest that light induces increased cabII-1 transcript abundance in two ways: 1) by virtue of its role in the light reactions of photosynthesis and 2) by a blue lightstimulated mechanism which is independent of photosynthesis. We have also examined the role of acetate in regulating cabII-1 mRNA levels in the dark. In both green- and yellow-in-the-dark strains, 15 mM Na-acetate, added to synchronized cells in the dark, induces an increase in cabII-1 mRNA abundance with a temporal accumulation pattern very similar to that induced by continuous white light. We suggest that by providing an energy source, acetate stimulates cellular growth, cell cycle progression, and increased cabII-1 mRNA abundance. Interestingly, in cells exposed to light, acetate inhibits the light-induced increase in cabII-1 mRNA abundance by a mechanism which is not yet understood.
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  • 79
    ISSN: 1573-5028
    Keywords: abscisic acid ; aleurone layer ; α-amylase ; gene expression ; gibberellic acid ; isozymes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The treatment of barley aleurone layers with gibberellic acid (GA3) results in the synthesis of two groups of α-amylase isozymes. Addition of abscisic acid (ABA) at the same time as GA3 inhibited the synthesis of both groups of isozymes. However, midcourse ABA addition (12 h or later after GA3) had a more inhibitory effect on the high pI α-amylase group than on the low pI α-amylase group. This midcourse inhibition was detectable within 2 h of ABA addition. Northern analysis results using cDNA probes for the high pI and low pI α-amylase groups paralleled the protein synthesis results for both isozyme groups. High pI α-amylase mRNA levels began to decrease within 2 h of midcourse ABA treatment and were less than 10% of the original level by 4 h. The levels of low pI α-amylase mRNA were decreased less by midcourse ABA addition than were high pI mRNA levels. Cordycepin and cycloheximide blocked the effects of midcourse ABA addition on α-amylase mRNA. These observations indicate that ABA inhibits α-amylase expression at the pretranslational level and that protein and RNA synthesis are required for midcourse ABA action to occur. Our results also show that α-amylase mRNA, which has been thought to be very stable, is degraded after midcourse ABA treatment.
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  • 80
    ISSN: 1573-5028
    Keywords: plant transformation ; CaMV promoters ; gene expression ; protein stability ; beta-conglycinin
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A gene encoding the α′-subunit of β-conglycinin was ligated to the 19S and 35S promoters of Cauliflower Mosaic Virus and introduced into petunia plants on a disarmed Ti-plasmid using Agrobacterium tumefaciens. Transformed cells were regenerated into whole plants and ummunoreactive polypeptides and hybridizable, polyadenylated mRNA were detected in transformed tissues. Expression from the 35S promoter was 10 to 50 times greater than expression from the 19S promoter. The level of immunodetectable polypeptides was greater in seeds than in leaves or callus tissue. In addition, the pattern of α′-polypeptide breakdown products was distinctive in seeds and leaves. We conclude that in seeds the higher levels of the α′-polypeptide reflect enhanced stability of this protein.
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  • 81
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    Plant cell, tissue and organ culture 8 (1987), S. 225-233 
    ISSN: 1573-5044
    Keywords: Brassica napus ; thin cell layer ; protoplasts ; plant regeneration ; rapeseed ; organogenesis ; callus
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cell layer strips composed of the epidermis and 7–9 layers of subepidermal cells were isolated from the 3–4 terminal internodes of Brassica napus cv Westar plants at the early flowering stage. The strips were precultured for one day in modified liquid MS [11] medium and subsequently incubated for 17–18 h in a 0.4 M mannitol solution containing 1% Macerozyme and 1% Cellulase ‘Onozuka’ R-10. Protoplast yield was 2–2.8×106 per 1.0g of tissue. Protoplasts were cultured at 1×105/ml in three different media: S1 [13], B [12] and L[8]. The first cell divisions occurred after 2–8 days of culture at frequencies of 20–54%. The highest growth rate of colonies was obtained in L medium containing 0.4 M sucrose and 2% Ficoll. After 4 weeks, green calli, 1–2 mm in diameter were transferred onto B5 [2] medium with 3 mgl-1 zeatin, 1% sucrose, 0.1 M mannitol and 0.5% agarose for shoot regeneration. Up to 20% of the calli regenerated shoots which subsequently were rooted and established in soil in the greenhouse.
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  • 82
    ISSN: 1573-5028
    Keywords: hypersensitive response ; Brassica campestris ; Xanthomonas campestris pv. vitians ; mRNA induction ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Xanthomonas campestris pv. vitians, a pathogen of lettuce, elicits a hypersensitive response within 12 hours of inoculation into Brassica leaves, characterized by tissue collapse, loss of membrane integrity, vein blockage and melanin production. In contrast, the compatible pathogen, X. c. pv. campestris, has no visible effects on leaves for 48 hours, after which inoculated areas show chlorosis which eventually spreads, followed by rotting. mRNA was prepared from leaves inoculated with suspensions of both pathovars or with sterile medium up to 24 hours following inoculation. In vitro translation of total and poly A+ RNA in rabbit reticulocyte lysate in the presence of 35S methionine followed by separation of the polypeptide products by 2D-PAGE, allowed comparison of the effects of these treatments on plant gene expression. Major changes in gene expression were observed as a consequence of the inoculation technique. In addition, after inoculation with X. c. vitians, up to fifteen additional major polypeptides appeared or greatly increased by four hours. Some of these had disappeared by nine hours and several more had appeared. No major polypeptides disappeared or decreased greatly in intensity following inoculation with X. c. vitians.
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  • 83
    ISSN: 1573-5028
    Keywords: coordinate regulation ; Cucurbita phytochrome cDNA ; gene expression ; light regulation ; multiple transcripts ; regulatory photoreceptor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have isolated several cDNA clones for phytochrome from a dicot, Cucurbita pepo L. cv. Black Beauty (zucchini), and have used them to study the regulation of Cucurbita phytochrome mRNA levels. A cDNA library was constructed from poly(A)+ RNA isolated from etiolated Cucurbita hypocotyl hooks and enriched for phytochrome mRNA by size fractionation. This library was screened with a 32P-labeled fragment isolated from an Avena phytochrome cDNA clone. Several putative phytochrome clones were isolated and mapped by restriction endonuclease analysis. On the basis of this analysis there is no evidence for the expression of multiple phytochrome genes in Cucurbita. Recent sequence analysis has confirmed that the largest of these clones, pFMD1 (∼3.6 kb), does indeed encode phytochrome and that it contains the entire amino acid coding sequence for Cucurbita phytochrome (33). RNA blot analysis has revealed that two polyadenylated phytochrome transcripts (∼5.6 kb and ∼4.2 kb) are present in both cotyledons and hypocotyl hooks of Cucurbita. In etiolated Cucurbita seedlings given a saturating pulse of red light, the abundance of both transcripts coordinately declines to 50–60% of the dark levels within 3 h and reaccumulates to dark levels within 24 h. Reversal of induction of this response by a far-red light pulse immediately following red light treatment is not observed, which is in contrast to the far-red reversibility of the red light promoted decrease in phytochrome mRNA abundance observed in Avena (6). Etiolated seedlings transferred to continuous white light also show a coordinate decrease in the levels of the two RNAs to ∼40% of the dark levels within 3 h. The magnitude of the light-induced decline in phytochrome mRNA abundance in Cucurbita is substantially less than the decrease previously reported for Avena (6).
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  • 84
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    Plant cell, tissue and organ culture 10 (1987), S. 187-196 
    ISSN: 1573-5044
    Keywords: protoplasts ; Azolla ; Sporophytes ; ferns ; Cellulysin ; Pectolyase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have developed a method for producing protoplasts from the heterosporous water fern Azolla using a combination of Cellulysin (4.0%) and Pectolyase (0.025%) in 0.6 M mannitol containing 6 mM CaCl2 2H2O. These protoplasts regenerate new cell walls within 48 hours when cultured on modified Gamborg B-5 medium.
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  • 85
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    Plant cell, tissue and organ culture 11 (1987), S. 179-188 
    ISSN: 1573-5044
    Keywords: Solanum melongena ; protoplasts ; lamina ; petioles ; stems
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Plants of Solanum melongena were propagated under in vitro conditions (27°C, 12h/day illumination at 62 μEm-2s-1, 60% humidity) by subculture of terminal and lateral cuttings on MS medium +20 gl-1 sucrose + Morel and Wetmore vitamins at 1/8 strength and 7 gl-1 agar. Lamina, petioles and stems of 3-week-old cuttings were used as sources of protoplasts. The best mean yield of protoplasts was obtained from the lamina with 9,030×103 protoplasts per gram of tissue. Petioles and stems yielded respectively 3,144×103 and 1,220.4×103 protoplasts per gram of tissue. first division of petiole and stem protoplasts occurred within 48 h, while lamina protoplasts underwent division after 3–4 days of culture in KM8p medium +2,4-D(0.2 gl-1) + zeatin (0.5 mgl-1) + NAA (1 mgl-1) and 0.35M glucose as osmoticum. The highest percentage of dividing cells was obtained from petiole material, estimated at 33.4% after 7 days, compared to 23.8% and 19.4% respectively for stem and lamina protoplasts. When BAP replaced zeatin in KM8p, the division percentage of lamina protoplasts was reduced to 10–15%. When transferred to regeneration medium, all calli derived from KM8p + zeatin formed deep-green spots identified as embryo-like structures, while only few calli from KM8p + BAP underwent shoot organogenesis without formation of green spots. Some of embryo-like structure developed into plantlets with a frequency of 1–2 plantlets per callus especially on MS medium + zeatin (4 mgl-1) + IAA (0.2 mgl-1). Maintaining protoplast-derived calli on MS + BAP (0.5 mgl-1) + NAA (0.5 mgl-1) for more than 3 weeks resulted in a decrease and loss of cell totipotency.
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  • 86
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    Cancer and metastasis reviews 6 (1987), S. 503-520 
    ISSN: 1573-7233
    Keywords: animal models ; chemotherapy ; gene expression ; metastasis ; tumour progression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Cancer chemotherapy is currently undergoing an intensive reappraisal because of its unimpressive performance against the major common cancers. There are a number of possible reasons for this lack of success; one considered here is that under some circumstances anti-neoplastic drug treatment actually increases the malignant behaviour of tumours. Support for this idea comes mainly from experimental studies in which drug treatments increased metastatic spread. Investigation of this phenomenon shows that drug induced modifications of the host, including immunosuppression and vascular damage, can indeed facilitate metastasis. In addition, new data are presented demonstrating that the direct action of drugs on the tumour cells themselves can have similar enhancing effects. The possible mechanisms underlying such direct effects are discussed and the ability of anti-cancer drugs to cause genetic mutations, amplify genes, and alter gene expression are considered. While the nature and extent of this facilitation of tumour malignancy is not fully understood, it is suggested that this possibility should be considered in the design of treatment protocols.
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  • 87
    ISSN: 1572-994X
    Keywords: EBV ; gene expression ; immortalization ; cDNA cloning ; Northern blot hybridization ; tonsil lymphocytes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract Two infection systems have been introduced in order to study viral gene expression at the initial period of Epstein-Barr virus (EBV) infection which leads to immortalization. The data indicate that major viral gene expression in tonsil lymphocytes at 2 days post-infection (p.i.) with EBV is very similar to that observed in latently infected cells. Both tonsil lymphocyte and BJAB cell (lymphoblastoid cells free of EBV genome) infection with EBV induced similar viral gene transcription. Twelve cDNA clones were prepared from poly(A) RNA of tonsil lymphocytes infected with EBV 2 days p.i. by hybridization with BamHI fragments of EBV DNA. Some cDNAs were derived from primary transcripts of the BamHI-WYHK region, suggestive of splicing of a large transcript. It is possible that a number of cDNA clones may be derived from cellular genes. The derivation of these cDNA clones is being studied.
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  • 88
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    Plant molecular biology 8 (1987), S. 251-264 
    ISSN: 1573-5028
    Keywords: chromatin structure ; DNase I hypersensitivity ; gene expression ; sucrose synthetase ; Zea mays
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The local chromatin structure of the Shrunken-1 (Sh) gene of maize was probed by analyzing DNase I hypersensitivity. Sh encodes the gene for sucrose synthetase, a major starch biosynthetic enzyme, which is maximally expressed in the endosperm during seed maturation. In addition to general DNase I sensitivity, specific DNase I hypersensitive sites were identified in endosperm chromatin that mapped near the 5′ end of the Sh gene. The pattern of hypersensitive sites and their relative sensitivity were altered in other non-dormant tissues that produce little or no enzyme. However, some changes in chromatin structure appear to be independent of Sh gene expression and may reflect general alterations associated with plant development. The chromatin structure of several sh mutations, induced by Ds controlling element insertions, was also analyzed. Although the insertions perturbed expression of the gene, there were no notable effects on local chromatin structure.
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  • 89
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    Plant molecular biology 8 (1987), S. 363-373 
    ISSN: 1573-5028
    Keywords: transformation ; MgCl2-PEG-electroporation ; competence ; protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Direct gene transfer into plant protoplasts has been recently developed, and conditions for high frequency transformation of SR1 tobacco protoplasts established. In this paper we analyse numerous transformation parameters in a comparative study on SR1 Nicotiana tabacum and N. plumbaginifolia, and report on a simple chemical technique for very efficient protoplast transformation. It is based on the synergistic interaction of MgCl2 and PEG. The technique yielded up to 1400 transformants per 3×105 treated N. tabacum protoplasts (up to 4.8% of the survivors, late selected clones). Using N. plumbaginifolia, the frequencies were 10-fold lower, indicating that the ‘competence’ for transformation has a species-specific component.
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  • 90
    ISSN: 1573-5028
    Keywords: Bradyrhizobium ; gene expression ; nodulin ; soybean
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract In vitro translation products of total RNA isolated from soybean nodules at successive stages of nodule development were analyzed by two-dimensional gel electrophoresis. In that way the occurrence of over 20 mRNAs specifically transcribed from nodulin genes was detected. The nodulin genes could be divided into two classes according to the time of expression during nodule development. Class A comprises at least 4 nodulin mRNAs which are found when a globular meristem is present in the root cortex. These class A nodulin genes have a transient expression. Class B nodulin genes are expressed when the formation of a nodule structure has been completed. Bradyrhizobium japonicum nod + fix-mutants, with large deletions spanning the nif H,DK region, still induced nodules showing normal expression of all nodulin genes, indicating that the nif H,DK region is not involved in the induction of nodulin genes. In nodules induced by Bradyrhizobium japonicum nod + fix-mutant HS124 the bacteria are rarely released from the infection thread and the few infected cells appear to be collapsed. All class A and class B nodulin genes are expressed in HS124 nodules with the exception of 5 class B genes.
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  • 91
    ISSN: 1573-5028
    Keywords: potato tuber ; elicitor ; arachidonic acid ; cDNA cloning ; gene expression ; PR proteins
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Accumulation of messenger RNAs in potato tuber discs was analysed during the hypersensitive response induced by treatment with the biotic elicitor arachidonic acid. In vitro translation of polysomal poly(A)+ RNAs indicated that the accumulation of some sixteen mRNAs varied following treatment with arachidonic acid, and that the level of thirteen of these was increased. Two cDNA closes (pSTH-1 and-2) were isolated from a library of elicitor-treated tissue cDNAs. Northern blot analysis using these clones as molecular probes indicated that the levels of at least two mRNAs were markedly increased after elicitor treatment. In hybrid-released translation experiments, each of the cDNA clones selected more than one mRNA. Translation of these mRNAs yielded two polypeptides of Mr 45 000 (for the pSTH-1 clone), and three polypeptides of Me 17 000 (for the pSTH-2 clone). The low molecular weight polypeptides may correspond to potato pathogenesis-related (PR) proteins.
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  • 92
    Electronic Resource
    Electronic Resource
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 34 (1987), S. 13-22 
    ISSN: 0730-2312
    Keywords: TPA ; tumor promotion ; JB6 cells ; transcriptional induction ; inducible mRNA ; 2ar mRNA ; mRNA induction ; serum-inducible ; JB6 ; TPA ; tumor promoter ; nuclear “run-off” transcription ; gene expression ; colony screening ; mRNA ; growth factors ; competence ; 3T3 fibroblasts ; protein kinase C ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: From the mouse JB6 epidermal cell line C122 we have isolated a cDNA clone representing a 1.6-kilobase mRNA, called 2ar, that exhibits a biphasic induction in response to 12-O-tetradecanoyl-phorbol-13-acetate (TPA). The first phase of induction in subconfluent cells is transient, peaking at 6 h after the addition of TPA and returning to noninduced levels by 24 h. When the cells reach plateau density, in the continued presence of TPA, this mRNA is reinduced and remains so upon continued exposure to the tumor promoter. Serum and certain growth factors also induce 2ar mRNA in serum-deprived quiescent fibroblasts. In vitro nuclear “run-on” transcription experiments indicate that the induction of 2ar mRNA is controlled at the transcriptional level.
    Additional Material: 5 Ill.
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  • 93
    ISSN: 0148-7280
    Keywords: immunofluorescence ; early development ; gene expression ; H-Y antigen ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: An indirect immunofluorescence assay was used to detect the presence of male-specific protein(s) on various stages of preimplantation porcine embryos. Embryos were collected at slaughter from the reproductive tracts of day-2.5, -4, -5, -6, and -8 (day 0 = first day of estrus) sows and gilts. Embryos were placed in medium containing an anti-male primary antibody, washed, and transferred to culture drops containing a fluorescein isothiocyanate (FITC)-labeled secondary antibody. Embryos were classified as either fluorescent (H-Y positive) or nonfluorescent (H-Y negative), transferred to coded drops, and karyotyped to examine sex chromosomes. A total of 91 eight-cell to blastocyst stage embryos were evaluated; of these, 46% were classified as fluorescent and 54% as nonfluorescent. Of readable metaphase spreads (65%) from these embryos, 81% (48 of 59, P 〈 0.005) were correctly sexed by immunological detection of the male-specific antigen. Although 13 % (2/15)of four-cell embryos evaluated were classified as fluorescent, the accuracy with which embryos at this stage were sexed by detection of H-Y antigen was not different from 50%. Fifty percent of eight-cell embryos were classified as H-Y positive with 78% of embryos correctly sexed. It was concluded that the eight-cell embryo is the earliest stage of development for which there is evidence for expression of H-Y antigen. Detection of the male-specific protein was difficult at the expanded blastocyst stage.
    Additional Material: 2 Tab.
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  • 94
    Electronic Resource
    Electronic Resource
    Chichester [u.a.] : Wiley-Blackwell
    Developmental Genetics 8 (1987), S. 125-133 
    ISSN: 0192-253X
    Keywords: retrovirus ; embryonal carcinoma ; embryonic gene ; DNA methylation ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Northern blot analysis and in vitro nuclear transcription assays were performed in order to clarify conflicting reports on the expression of intracisternal A particle (IAP) genes in embryonal carcinoma (EC) cell lines. Results demonstrate that post-transcriptional mechanisms control the final steady-state levels of IAP RNA in EC cells. IAP genes were further found to be undermethylated in IAP-expressing EC cell lines.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
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  • 95
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 3 (1987), S. 149-160 
    ISSN: 0749-503X
    Keywords: DNA repair ; RAD2 ; Saccharomyces ; gene expression ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The cloned RAD2 gene of S. cerevisiae was tailored into regulatable expression vectors for overexpression of Rad2 protein in E. coli and in yeast. In E. coli both Rad2/β-galactosidase fusion protein and native Rad2 protein are insoluble, but are extractable with 1% Sarkosyl. In yeast some of the overexpressed native Rad2 protein is also insoluble; however, soluble protein is readily detected by immunoblotting with Rad2-specific antibodies. All forms of the protein detected in transformed or untransformed yeast cells and the insoluble species in E. coli migrate in denaturing polyacrylamide gels with an apparent molecular weight considerably larger than the size predicted from the sequence of the RAD2 coding region. This property is not the result of post-translational glycosylation detectable by binding of concanavalin A, or of phosphorylation of the protein. Overexpression of the RAD2 gene is toxic to yeast. Transformed yeast cells grow much more slowly than untransformed controls and when yeast transformants are serially propagated cultures show considerable colony heterogeneity and concomitant selection for rapidly growing variants which express less Rad2 protein. Antisera raised against Rad2/β-galactosidase fusion protein expressed in E. coli do not cross-react with Rad1, Rad3 or Rad10 protein in crude extracts of yeast, nor with purified E. coli UvrA, UvrB or UvrC proteins.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
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  • 96
    ISSN: 1573-4919
    Keywords: cell cycle ; gene expression ; oncogenes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: Summary A number of genes and cDNA sequences (including at least four oncogenes) are known to be expressed in a cell cycle-dependent manner, i.e. the levels of specific mRNAs vary with the phases of the cell cycle. In order to explore the significance of some of these sequences in the mitogenic response, we have investigated the expression of 8 cell cycle-dependent sequences (plus two control sequences, not expressed in a cell cycle-dependent manner) under a variety of conditions. These conditions included cells of different types, from different species, stimulated to proliferate by different mitogens. The genes (or sequences) studied included five cDNA clones whose sequences are preferentially expressed in early G1, i.e. two cDNA clones inducible by platelet-derived growth factor (JE-3 and KC-1), and three cDNA clones inducible by serum (2A9, 2F1, 4F1); and three oncogenes (c-myc, c-rasHa and p53) whose expression is known to be cell cycle-dependent. All of the tested genes, except 2A9, c-rasHa and the control genes, are expressed in a cell cycle-dependent manner in human peripheral blood mononuclear cells stimulated by phytohemagglutinin and in serum-stimulated mouse and Syrian hamster fibroblasts. The inducibility of these genes by different mitogens in cells of different types and from different species strongly suggests that these genes play a role in cell cycle progression. This conclusion is further supported by the known structural and functional similarities between cell-cycle dependent genes, oncogenes and genes coding for cell-cycle related molecules.
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  • 97
    Electronic Resource
    Electronic Resource
    Springer
    Bioscience reports 6 (1986), S. 621-631 
    ISSN: 1573-4935
    Keywords: brown adipose tissue ; thermogenin ; uncoupling protein ; gene expression ; adrenergic effects
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Abstract By the use of an earlier characterised cDNA clone, CIN-1, corresponding to a sequence of the mRNA coding for the brown-fat specific “uncoupling” protein, thermogenin, the amount of thermogenin mRNA found in the brown adipose tissue of mice was quantitatively investigated under different physiological and pharmacological conditions. It was found that a 4 hr cold stress led to a 7-fold increase in the amount of thermogenin mRNA; injection of norepinephrine had a significant but smaller effect. Most notably, isoprenaline (β-agonist) and phenylephrine (α-agonist) had in themselves no effect, but when injected together were able to increase the mRNA level synergistically. In 4 hr cold-stressed mice, norepinephrine, isoprenaline and cholera toxin could all further potentiate the effect of the cold stress itself on the mRNA level. Insulin and the glucocorticoid dexamethasone both had weak stimulatory effects on the mRNA level. It is concluded that an increase in intracellular cAMP levels is a necessary and perhaps sufficient stimulus for the increase in thermogenin gene expression. However, at least underin vivo conditions, this increase requires stimulation of both α- andβ-adrenergic pathways.
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  • 98
    ISSN: 1573-5079
    Keywords: carbon dioxide fixation ; freezing stress ; photophosphorylation ; photosynthetic electron transport ; protoplasts ; Valerianella locusta
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mesophyll protoplasts were isolated from unhardened and cold-acclimated leaves of Valerianella locusta L. and subjected to freeze-thaw treatment. To evaluate the extent and course of freezing injury, photosynthetic reactions of whole protoplasts and of free thylakoid membranes, liberated from protoplasts by osmotic lysis, were measured. In addition, the integrity of the protoplasts was determined by microscopy. The results reveal an increased frost tolerance of protoplasts isolated from acclimated leaves with respect to all parameters measured. CO2-dependent O2 evolution (representing net photosynthetic CO2 fixation of protoplasts) was the most freezing-sensitive reaction; its inhibition due to freeze-thaw treatment of protoplasts was neither correlated with disintegration of the plasma membrane, nor was it initiated by inactivation of the thylakoid membranes. The frost-induced decline of protoplast integrity was not closely correlated to thylakoid damage either. Freezing injury of the thylakoid membranes was manifested by inhibition of photosynthetic electron transport and photophosphorylation. Both photosystems were affected by freezing and thawing with strongest inhibition occurring in the water-oxidation system or at the oxidizing site of photosystem II. Photophosphorylation responded more sensitively to freezing stress than electron transport, although uncoupling (increased permeability of the thylakoid membranes to protons) was not a conspicuous effect. The data are discussed in relation to freezing injury in leaves and seem to indicate that frost damage in vivo is initiated at multiple sites.
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  • 99
    Electronic Resource
    Electronic Resource
    Springer
    Plant molecular biology 7 (1986), S. 51-61 
    ISSN: 1573-5028
    Keywords: Soybean ; Rhizobium ; nitrogen fixation ; gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Nodulins are organ-specific plant proteins induced during symbiotic nitrogen fixation. Nodulins play both metabolic and structural roles within infected and uninfected nodule cells. In soybean, several nodulin genes, coding for abundant nodulins, have been identified and isolated. Structural analysis of some of these genes has revealed their possible mode of regulation and the subcellar location of the protein product. Studies of ineffective symbiosis based on cultivar-strain genotype differences suggested that both partners influence the expression of nodulin genes. Concomitant with nodule organogenesis, the Rhizobium undergoes substantial differentiation leading to the accumulation of nodule-specific bacterial proteins, bacteroidins. The major structural alteration occuring in the infected cell is the formation of a membrane enclosing the bacteroid (peribacteroid membrane). A number of nodulins are specifically targetted to this membrane during endosymbiosis. The induction of nodulins and bacteroidins leads to the formation of an effective nodule. Nodulin genes can be induced in vitro by factors derived from nodules suggesting that trans-activators may be involved in derepression of the host genes necessary for Rhizobium-legume symbiosis.
    Type of Medium: Electronic Resource
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  • 100
    Electronic Resource
    Electronic Resource
    Springer
    Plant cell, tissue and organ culture 6 (1986), S. 47-59 
    ISSN: 1573-5044
    Keywords: cotton ; ovule epidermis ; protoplasts ; cell wall regeneration ; β-1,3-glucans ; β-1,4-glucans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Culture protocols were developed and characterization of the regenerated cell walls was performed for protoplasts of cotton (Gossypium hirsutum L., L., var. Acala SJ-2) ovule epidermal cells. This work was undertaken in order to extend studies concerning nutritional effects and regulation of nucleotide sugar incorporation into β-1,3- and β-1,4-glucan components of cotton fiber cell walls. Protein and carbohydrate polymers and recovered from the culture medium. Analysis of a cellular fraction indicated that the majority of 14C incorporated from [14C] glucose was present in the hot-water-soluble fraction of the cells. The majority of label incorporated into cell wall material could be solubilized with acetic-nitric reagent, indicative of noncellulosic material, and characterized as β-1,3-linked glucans. Only 5 to 15% of the regenerated cell wall could be characterized as β-1,4-linked glucose indicative of cellulose.
    Type of Medium: Electronic Resource
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