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  • 1
    Electronic Resource
    Electronic Resource
    DNA sequence of the metC gene and its flanking regions from Salmonella typhimurium LT2 and homology with the corresponding sequence of Escherichia coli (1989)
    Springer
    Molecular genetics and genomics 216 (1989), S. 164-169 
    Details
    ISSN: 1617-4623
    Keywords: metC ; Cystathionine-β-lyase ; Nucleotide sequence ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The DNA sequence of the Salmonella typhimurium metC gene and its flanking regions was determined. The metC gene contains an open reading frame of 1185 nucleotides encoding a polypeptide of 395 amino acids with a predicted molecular weight of 42874 daltons. S1 nuclease mapping experiments located the transcription start site of the metC gene. The nucleotide sequence and the deduced amino acid sequence for the metC genes of S. typhimurium and Escherichia coli were compared. Although there are 279 nucleotide replacements, most do not change the amino acid sequence. Nucleotide sequence analysis of the flanking regions of the S. typhimurium metC gene shows that there is an open reading frame upstream and an open reading frame downstream of the gene. The existence of the divergently transcribed upstream open reading frame (designated ORF1) was confirmed by the construction of an ORF1-lacZ fusion. The transcription start site of ORF1 was determined by S1 nuclease mapping.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00332246
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  • 2
    Electronic Resource
    Electronic Resource
    Cloning and sequence analysis of the 18 S ribosomal RNA gene of tomato and a secondary structure model for the 18 S rRNA of angiosperms (1989)
    Springer
    Molecular genetics and genomics 219 (1989), S. 17-25 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Small subunit ; Ribosomal DNA ; Sequence comparison ; Lycopersicon esculentum
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The gene of a cytoplasmic 18 S ribosomal RNA (18 S rDNA) of the dicotyledonous plant tomato (ycopersicon esculentum) cv. Rentita has been cloned, and its complete primary structure has been determined. The tomato 18 S rDNA is 1805 by long with a G+C content of 49.6%. Its sequence exhibits 94%–96% positional identity when it is colinearly aligned with the previously reported sequences of the 17–18 S rDNAs of the dicot soybean and the monocots maize and rice. A model of the secondary structure of the 18 S rRNA of angiosperms is presented and its genera-specific structural features are compared with a current eukaryotic 18 S rRNA consensus model.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00261152
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  • 3
    Electronic Resource
    Electronic Resource
    Stable-light-emitting Escherichia coli as a biosensor (1989)
    New York : Wiley-Blackwell
    Journal of Bioluminescence and Chemiluminescence 4 (1989), S. 551-554 
    Details
    ISSN: 0884-3996
    Keywords: Recombinant DNA ; luminescence ; luciferase gene ; Vibrio harveyi ; toxic substances ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: We have studied possibilities for constructing Escherichia coli strains capable of producing stable light. Light production in E. coli is achieved by cloning the genes encoding bacterial luciferase from Vibrio harveyi. To gain the advantage of sensitive detection of light we transferred the genes under the control of a strong, regulatable promoter system.Stabilization of light produced by E. coli clones was accomplished by finding the optimal plasmid construction and growth conditions as well as suitable measuring buffers. The adjustment of the luciferase synthesis for bioluminescence measurements to a high but not harmful level gives healthy cells and stable luciferase. Cultivation at 30 °C in an uninduced state was found to be the most important factor in getting stable-light production. The overall cell metabolism being unstressed gives us the possibility of monitoring cell physiology and factors affecting it via bioluminescence reactions in vivo. To make the results easy to interpret the light emission has to be stable during a measurement period of one to several hours. In the case of the original light-producing bacteria, Vibrio and Photobacterium strains it has not thus far been possible to find conditions where light emission would be stable for several hours. Based on our findings an automated biosensor system can be developed to monitor the effects of biologically active compounds against stable-light-producing bacteria.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bio.1170040172
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  • 4
    Electronic Resource
    Electronic Resource
    Evaluation of the stability of human erythropoietin in samples for radioimmunoassay (1988)
    Springer
    Journal of molecular medicine 66 (1988), S. 241-245 
    Details
    ISSN: 1432-1440
    Keywords: Erythropoietin ; Stability ; Radioimmunoassay ; Polycythemic mouse bioassay ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Radioimmunoassays for erythropoietin are limited so far to a few specialized laboratories and this requires transport and storage of samples. We therefore tested the stability of immunoreactive erythropoietin in serum and plasma samples obtained from a uremic and a nonuremic anemic patient. No significant change in the concentration of immunoreactive erythropoietin was found in either serum or plasma samples for up to 14 days of storage. This type of stability was observed no matter whether the samples were stored at room temperature, 4° C, or −20° C. There was no difference between the estimates of erythropoietin in serum and heparinized plasma. Validity of the radioimmunoassay used in this study was demonstrated by parallelism of dilution curves of test specimens and the 2nd International Reference Preparation for erythropoietin and by a close correlation between the immunoreactivity and the bioactivity of the hormone, as assessed in the same samples by the exhypoxic polycythemic mouse bioassay. In conclusion the data obtained clearly indicate that the necessity of storage and transport of clinical samples does not limit the practicability of the radioimmunoassay for erythropoietin.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01748163
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  • 5
    Electronic Resource
    Electronic Resource
    Human cytomegalovirus DNA in the temporal cortex of a schizophrenic patient (1988)
    Springer
    European archives of psychiatry and clinical neuroscience 238 (1988), S. 110-113 
    Details
    ISSN: 1433-8491
    Keywords: Schizophrenia ; Cytomegaloviruses ; Virus diseases ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Using highly sensitive nucleic acids hybridization techniques, which allow the detection of 0.1–0.5 single copy gene equivalents per cell, DNA from the temporal cortex of seven definite schizophrenics, five persons with schizophrenia-like psychoses, three patients with Huntington's chorea and nine mentally normal individuals were probed with human cytomegalovirus (HCMV) DNA. A clear hybridization signal was obtained with DNA from the temporal lobe of a young schizophrenic patient, whereas DNA from the temporal cortex of controls did not hybridize to the HCMV probe. This finding is in agreement with the cytomegalovirus hypothesis of schizophrenia and hints at the possibility that viral infection of the temporal cortex may in some sporadic cases be a contributing factor to the development of schizophrenic psychoses. There is no indication, however, that infection of the central nervous system with HCMV is an aetiological factor in the great majority of schizophrenic disorders. Clearly further studies, preferably in situ hybridizations of whole brains, are needed to prove or disprove the cytomegalovirus hypothesis of schizophrenia.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00452786
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  • 6
    Electronic Resource
    Electronic Resource
    Cloning and nucleotide sequence of the Salmonella typhimurium LT2 metF gene and its homology with the corresponding sequence of Escherichia coli (1988)
    Springer
    Molecular genetics and genomics 212 (1988), S. 246-251 
    Details
    ISSN: 1617-4623
    Keywords: metF ; 5,10-methylenetetrahydrofolate reductase ; Recombinant DNA ; Nucleotide sequence
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The Salmonella typhimurium LT2 metF gene, encoding 5,10-methylenetetrahydrofolate reductase, has been cloned. Strains with multicopy plasmids carrying the metF gene overproduce the enzyme 44-fold. The nucleotide sequence of the metF gene was determined, and an open reading frame of 888 nucleotides was identified. The polypeptide deduced from the DNA sequence contains 296 amino acids and has a molecular weight of 33 135 daltons. Mung bean nuclease mapping experiments located the transcription start point and possible transcription termination region for the gene. There is a 25bp nucleotide sequence between the translation termination site and the possible transcription termination region. This region possesses a GC-rich sequence that could form a stable stem and loop structure once transcribed (ΔG=-9 kcal/mol), followed by an AT-rich sequence, both of which are characteristic of rho-independent transcription terminators. The nucleotide and deduced amino acid sequences of the S. typhimurium metF gene are compared with the corresponding sequences of the Escherichia coli metF gene. The nucleotide sequences show 85% homology. Most of the nucleotide differences found do not alter the amino acid sequences, which show 95% homology. The results also show that a change has occurred in the metF region of the S. typhimurium chromosome as compared to the E. coli chromosome.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00334692
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  • 7
    Electronic Resource
    Electronic Resource
    Cloning, sequencing and expression of a Bacillus bacteriolytic enzyme in Escherichia coli (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 241-248 
    Details
    ISSN: 1617-4623
    Keywords: Autolysin ; Hydrolase ; In vitro transcription and translation ; Recombinant DNA ; Transcriptional and translational signals
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Several hundred bacterial isolates were screened for bacteriolytic activity by growing them on agar medium containing autoclaved, lyophilized Micrococcus lysodeikticus cells as the substrate. A Bacillus sp. producing the largest lytic zone was selected. A genomic bank of this selected bacterium was constructed in the multi-functional vector pTZ18R, with partial SauIIIA DNA fragments inserted at the SalI restriction site. Screening of 800 colonies of this bank for cell lysis gave 5 recombinants exhibiting lytic activity, as detected by analysis of extracts of sonicated Escherichia coli cells on denaturing polyacrylamide gels containing autoclaved, lyophilized M. lysodeikticus cells as the substrate. One clone (pBH2500), expressed inE. coli strain NM522, was found to code for a lytic enzyme corresponding, in molecular weight, to the 27 kDa Bacillus sp hydrolase. This clone with an insertion of 2.5 kb was then subcloned as a 929 bp EcoRI-SauIIIA fragment in pTZ18R (pBH929) and showed higher cell lytic activity. A unique open reading frame for a protein of 251 amino acids, followed by a putative terminator sequence, was found after a consensus ribosome binding site. A putative leader sequence was identified in the first 37 amino acids. One truncated subclone (pBH703), corresponding to 196 out of 251 residues from the protein N-terminal end, still possessed lytic activity.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00337717
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  • 8
    Electronic Resource
    Electronic Resource
    Characterization of genetic transformation in Streptococcus oralis NCTC 11427: Expression of the pneumococcal amidase in S. oralis using a new shuttle vector (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 53-57 
    Details
    ISSN: 1617-4623
    Keywords: Competence ; Autolysins ; Choline ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have worked out conditions for the study of competence development and genetic transformation in Streptococcus oralis NCTC 11427 (type strain), a species that contains choline in the cell wall. The peak of competence was found at the early exponential phase of growth and the optimal conditions for transformation were achieved with shuttle plasmids prepared from S. pneumoniae or from Escherichia coli serving as donor DNA. Transformation with dye-bouyant density gradient purified plasmid preparations followed first-order kinetics. The pneumococcal amidase can be expressed in S. oralis harbouring a plasmid carrying the lytA gene. This enzyme lysed the cell wall of the transformed cell in the presence of detergents.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331302
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  • 9
    Electronic Resource
    Electronic Resource
    The repressor gene (c) of the Streptomyces temperate phage ϕc31: Nucleotide sequence, analysis and functional cloning (1988)
    Springer
    Molecular genetics and genomics 213 (1988), S. 269-277 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Helix-turn-helix motifs ; In vitro transcription-translation ; Phage immunity ; Exonuclease III deletions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The nucleotide sequence of the 3.4 kb SphI-G fragment that contained the repressor gene (c) of the temperate Streptomyces phage ϕc31 was determined. Analysis of this sequence revealed a large open reading frame with protein coding character and sequence changes in c gene point and deletion mutants identified this as the coding region of the repressor. Two of the mutants studied had undergone deletions of 1.1 kb and 1.4 kb that had occurred across short direct repeats of 6 bp and 11 bp, respectively. Coupled in vitro transcription-translation experiments using the cloned SphI-G fragment and Streptomyces lividans cell free extracts identified a protein product of approximately 72 kDa, in close agreement with that predicted from the nucleotide sequence. A strongly predicted helix-turn-helix motif that may be involved in DNA binding occurred towards the carboxy-terminus of the amino acid sequence. Initial attempts to clone the SphI-G fragment in Streptomyces failed; using information gained from the sequence analysis a smaller segment of this DNA fragment was cloned in S. lividans and conferred immunity to a clear plaque mutant (c1) of ϕc31.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00339591
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  • 10
    Electronic Resource
    Electronic Resource
    Secretion and export of IGF-1 in Escherichia coli strain JM101 (1988)
    Springer
    Molecular genetics and genomics 215 (1988), S. 19-25 
    Details
    ISSN: 1617-4623
    Keywords: IGF-1 ; Escherichia coli secretion/export ; LamB leader peptide ; Heterologous gene expression ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The processing of LamB-IGF-1 fusion protein and the export of processed IGF-1 (insulin-like growth-factor-1) into the growth medium was examined in the Escherichia coli host strain, JM101. Several strain or plasmid modifications were tried to increase export of periplasmic (Processed) IGF-1 into the growth medium of JM101. These included: (1) use of a lon null mutant strain to increase accumulation levels of unprocessed LamB-IGF-1 fusion protein; (2) use of an alternative drug resistance marker on the expression plasmid rather than beta-lactamase, thereby reducing any competition for processing of LamB-IGF-1 by signal peptidase; (3) examination of whether phage M13 gene III protein expression caused more periplasmic IGF-1 to be exported into the growth medium due to increased outer membrane permeability; and (4) examination of the effect of E. coli or yeast optimized IGF-1 codons. None of these strain or plasmid modifications caused any significant increase in export of IGF-1 into the growth medium of JM101. Solubility studies of LamB-IGF-1 and processed IGF-1 showed that virtually all of the LamB-IGF-1 and IGF-1 remaining within the cell after a 2 h induction period was insoluble. This implied that only soluble LamB-IGF-1 was processed to IGF-1 and that only soluble IGF-1 was exported into the growth medium. Taken together, the results indicated that LamB-IGF-1 and IGF-1 solubility were the limiting factors in secretion of IGF-1 into the periplasm and export of IGF-1 into the growth medium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331297
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  • 11
    Electronic Resource
    Electronic Resource
    A multigene family in Calothrix sp. PCC 7601 encodes phycocyanin, the major component of the cyanobacterial light-harvesting antenna (1988)
    Springer
    Molecular genetics and genomics 211 (1988), S. 296-304 
    Details
    ISSN: 1617-4623
    Keywords: Complementary chromatic adaptation ; Gene expression ; Photoregulation ; Phycobilisome ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In cyanobacteria, light is harvested by phycobilisomes which are essentially made up of chromophoric proteins called phycobiliproteins. We have characterized two gene clusters (cpcB1, cpcA1 and cpcB3, cpcA3) each encoding the two subunits of phycocyanin (βPC and αPC, respectively), one of the major phycobiliproteins in Calothrix 7601. Downstream from the gene encoding the PCα subunit in cluster 1, an open reading frame was found, cpcE1. These genes are organized in two transcriptional units, namely: cpcB3 A3 and cpcB1 A1 E1. All these genes are transcribed whatever the chromatic light received during cell growth. Consequently, although only one type of “constitutive” PC has been biochemically characterized, we have demonstrated that there are two cpc operons “constitutively” transcribed in this strain. With the previously described red light “inducible” cpcB2 A2 operon, there are three copies of the PC encoding genes in Calothrix 7601. The significance of this newly described multigene family in cyanobacteria is discussed. We have also mapped the 5′ and 3′ termini of the major transcript from the cpc1 operon. Analysis of the 5′ untranslated region of this transcript has revealed alternative secondary structures which are proposed to play a role in the regulation of the expression of this operon.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00330607
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  • 12
    Electronic Resource
    Electronic Resource
    Cloning of two chloramphenicol acetyltransferase genes from Clostridium butyricum and their expression in Escherichia coli and Bacillus subtilis (1988)
    Springer
    Molecular genetics and genomics 214 (1988), S. 328-332 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Chloramphenicol resistance ; cat gene ; Plasmids pC194 and pUB110 ; Inducible gene expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Two non-homologous chloramphenicol (Cm) acetyltransferase (CAT) genes, designated catA and catB, were cloned from Clostridium butyricum type strains and characterized by restriction mapping. Both genes are efficiently expressed in Escherichia coli and Bacillus subtilis. In contrast to analogous genes from staphylococci and bacilli, gene expression is not dependent on induction by Cm. The genes are considered as chromosomal, since no association with endogenous plasmids was detectable. Southern hybridization revealed a homology between catA and the staphylococcal Cm resistance plasmid, pC194. The subunit size of the clostridial CAT enzymes expressed in E. coli was determined as 22.5 kDa (catA) and 24 kDa (catB), respectively. The C. butyricum cat genes provide potentially useful selection markers for the construction of cloning vectors from cryptic clostridial plasmids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00337731
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  • 13
    Electronic Resource
    Electronic Resource
    Coordinated regulation of type I and type III collagen production and mRNA levels of pro α1(I) and pro α2(I) collagen in cultured morphea fibroblasts (1987)
    Springer
    Archives of dermatological research 279 (1987), S. 154-160 
    Details
    ISSN: 1432-069X
    Keywords: Collagen ; Fibroblasts ; Scleroderma ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Fibroblast cultures were started from affected and unaffacted skin areas of six patients with localized scleroderma in an active stage. The cell lines were studied for synthesis of procollagens and fibronectin by metabolic labeling with 3H-proline and for their contents of mRNAs for pro α1(I) and pro α2(I) collagen. For this purpose a cDNA clone for human pro α1(I) collagen mRNA was constructed. The clone was identified by restriction site mapping and hybridization to the specific mRNAs. All the scleroderma fibroblast lines produced increased amounts of type I and type III collagens and fibronectin during the early passages. The cell lines gradually reduced their elevated synthesis of collagen and fibronectin to normal or near normal levels by the tenth passage. The ratios of α1(I) and α2(I) chains and of type I and type III collagens, and the extent of type I procollagen processing, remained relatively unchanged in all the cultures. The cellular levels of type I procollagen mRNAs were increased in all the cells exhibiting an increased synthesis of collagen. The results suggest that in localized scleroderma the fibroblasts have undergone a coordinated activation of collagen synthesis at transcriptional level.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00413250
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  • 14
    Electronic Resource
    Electronic Resource
    Factors affecting the expression of foreign proteins inEscherichia coli (1987)
    Springer
    Journal of industrial microbiology and biotechnology 1 (1987), S. 277-282 
    Details
    ISSN: 1476-5535
    Keywords: Recombinant DNA ; Gene expression ; Genetic engineering ; Biotechnology
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary A variety of factors affect the expression of foreign proteins inEscherichia coli. These include: promoter strength, efficiency of ribosome binding, stability of the foreign protein inE. coli, location of the foreign protein inE. coli, the codons used to encode the foreign protein, the metabolic state of the cell, and the location, stability and copy number of the foreign gene. This paper contains a critical review of these factors with the idea that a detailed understanding of them is the key to the development of strategies for the efficient large-scale production of foreign proteins inE. coli.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569305
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  • 15
    Electronic Resource
    Electronic Resource
    The isolation of specific genes from the basidiomycete Schizophyllum commune (1987)
    Springer
    Current genetics 12 (1987), S. 547-554 
    Details
    ISSN: 1432-0983
    Keywords: Schizophyllum commune ; Transformation ; Gene isolation ; Basidiomycetes ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have developed a routine way to isolate genes directly from the basidiomycete fungus, Schizophyllum commune. Plasmid DNA from a genomic gene library was used to isolate five specific genes by complementation of Schizophyllum mutations via transformation. The mutant strains were deficient in the ability to synthesize either adenine (ade2 and ade5), uracil (ural, encoding orotidine-5′-phosphate decarboxylase; OMPdecase), tryptophan (rpl, encoding indole-3-glycerol phosphate synthetase; IGPS) or para aminobenzoic acid (pab1). In each case, Southern analysis revealed that transformation to prototrophy was concomitant with the integration of vector sequence into the genome of the S. commune mutant. Total DNA from transformants was restricted, religated, and used to transform E. coli. Ampicillin resistant plasmids were recovered from E. coli and tested for their ability to transform the corresponding mutant of S. commune. Plasmids complementing the ade2, adeS, pabl, trpl, and ural mutations were recovered.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00419565
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  • 16
    Electronic Resource
    Electronic Resource
    Fermentation of recombinant yeast producing hepatitis B surface antigen (1987)
    Springer
    Journal of industrial microbiology and biotechnology 2 (1987), S. 117-121 
    Details
    ISSN: 1476-5535
    Keywords: Fermentation ; Recombinant DNA ; Hepatitis B surface antigen
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Fermentations were performed to determine parameters affecting the expression of hepatitis B surface antigen (HBsAg) in the yeastSaccharomyces cerevisiae containing the HBsAg gene. These studies emphasized inereasing both the relative abundance (HBsAg: cell mass) and total production of HBsAg. Specific activity was increased 70-fold when cells were grown in shake flasks containing nonselective rather than selective medium. The addition of adenine, ammonium sulfate or glucose to the complex medium reduced the production of antigen. Results similar to those achieved in shake flasks were obtained when the growth was performed in fermenters. A nutrient addition system was employed to increase the production of cells and HBsAg. The addition of glucose to the culture medium increased cell mass 6-fold but decreased the production of antigen. This imbalance was corrected by supplementing the glucose with complex nutrients.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01569510
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  • 17
    Electronic Resource
    Electronic Resource
    Molecular cloning of the plant plasma membrane H+-ATPase (1987)
    Springer
    Plant and soil 99 (1987), S. 185-196 
    Details
    ISSN: 1573-5036
    Keywords: Antibody ; H+-ATPase ; Membrane ; Recombinant DNA ; Transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Mineral transport across the plasma membrane of plant cells is controlled by an electrochemical gradient of protons. This gradient is generated by an ATP-consuming enzyme in the membrane known as a proton pump, or H+-ATPase. The protein has a catalytic subunit of Mr=100,000 and is a prominent band when plasma membrane proteins are analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We generated specific rabbit polyclonal antibody against the Mr=100,000 H+-ATPase and used the antibody to screen λgtll expression vector libraries of plant DNA. Several phage clones producing immunoreactive protein, and presumably containing DNA sequences for the ATPase structural gene, were isolated and purified from a carrot cDNA library and a Arabidopsis genomic DNA library. These studies represent our first efforts at cloning the structural gene for a plant plasma membrane transport protein. Applicability of the technique to other transport protein genes and the potential for use of recombinant DNA technology in plant mineral transport research are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02370165
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  • 18
    Electronic Resource
    Electronic Resource
    The Saccharomyces cerevisiae MET3 gene: Nucleotide sequence and relationship of the 5′ non-coding region to that of MET25 (1987)
    Springer
    Molecular genetics and genomics 210 (1987), S. 307-313 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Methionine metabolism ; Negative control ; Regulatory regions
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In Saccharomyces cerevisiae, the expression of several genes implicated in methionine biosynthesis is coregulated by a specific negative control. To elucidate the molecular basis of this regulation, we have cloned two of these genes, MET3 and MET25. The sequence of MET25 has already been determined (Kerjan et al. 1986). Here, we report the nucleotide sequence of the MET3 gene along with its 5′ and 3′ flanking regions. Plasmids bearing different deletions upstream of the transcribed region of MET3 were constructed. They were introduced into yeast cells and tested for their ability to complement met3 mutations and to respond to regulation by exogenous methionine. The regulatory region was located within a 100 bp region. The sequence of this regulatory region was compared with that of MET25. A short common sequence which occurs 250–280 bp upstream of the translation initiation codon of the gene was found. This sequence is a good candidate for the cis-acting regulatory element.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00325699
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  • 19
    Electronic Resource
    Electronic Resource
    Cloning of two type II methylase genes that recognise asymmetric nucleotide sequences: FokI and HgaI (1987)
    Springer
    Molecular genetics and genomics 209 (1987), S. 570-574 
    Details
    ISSN: 1617-4623
    Keywords: Recombinant DNA ; Methylase ; Plasmid vector ; Gel electrophoresis ; Clearage map
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The modification genes of Flavobacterium okeanokoites and Haemophilus galinarum have been cloned into the vector pBR322 and expressed in Escherichia coli cells. FokI methylase gene is contained on a 3.80 kb piece of F. okeanokoites DNA. Plasmid constructs carrying this fragment of DNA are resistant to digestion by FokI restriction endonuclease but are sensitive to cleavage by HindIII, EcoRI and PstI. Unmodified λ DNA molecules, exposed in vitro to cell extracts prepared from cells habouring this plasmid, became resistant to digestion by FokI. The smallest HgaI methylase clone carries the pBR322 plasmid containing a 3.50 kb piece of H. galinarum DNA. This plasmid is resistant to digestion by HgaI. Neither the FokI nor the HgaI restriction endonuclease was detected in either clone. This is the first report of cloning modification genes whose protein products recognise asymmetric nucleotide sequences.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331164
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  • 20
    Electronic Resource
    Electronic Resource
    Molecular cloning of a ribosomal protein gene from the fission yeast Schizosaccharomyces pombe (1986)
    Springer
    Current genetics 10 (1986), S. 365-370 
    Details
    ISSN: 1432-0983
    Keywords: Recombinant DNA ; Cross hybridization ; 2D-electrophoresis ; Hybrid selection translation ; Transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Using the structural gene for the ribosomal protein L3 from Saccharomyces cerevisiae as a probe, we isolated a homologous fragment from genomic DNA of Schizosaccharomyces pombe. Analysis of the plasmid carrying this fragment by hybridization selection and 2D-electrophoresis revealed a 31 kDa ribosomal protein. Transformation of the vector pDB248x containing this fragment into Schizosaccharomyces pombe leads to an increased level of mRNA suggesting that we have cloned the entire and actively transcribed gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00418408
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  • 21
    Electronic Resource
    Electronic Resource
    Large mitochondrial genome and mitochondrial DNA size polymorphism in the mosquito parasite, Romanomermis culicivorax (1986)
    Springer
    Current genetics 11 (1986), S. 71-77 
    Details
    ISSN: 1432-0983
    Keywords: Mitochondrial DNA ; Population heterogeneity ; Molecular evolution ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Physical characterization of the mitochondrial genome derived from the obligate mosquito parasite, Romanomermis culicivorax has generated some surprising physical properties regarding the molecular structure of nematode mitochondrial DNA (mtDNA). Restriction enzyme analysis of this mtDNA has revealed a mitochondrial genome size of approximately 26 kb, the largest metazoan mtDNA reported to date. Isofemale lineages are monomorphic for one of three size variants, differing by 500-1,000 base pairs, present in our original field population. Cloned hybridization probes derived from a single region exhibiting a 600 by size polymorphism share strong homology with several spatially separated sites distributed about the mtDNA. This suggests that the homology is a result of repeated DNA sequence elements contained within this mitochondrial genome that contribute to mtDNA size polymorphism.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00389428
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  • 22
    Electronic Resource
    Electronic Resource
    Gene cloning in Aspergillus nidulans: isolation of the isocitrate lyase gene (acuD) (1986)
    Springer
    Molecular genetics and genomics 202 (1986), S. 271-275 
    Details
    ISSN: 1617-4623
    Keywords: Aspergillus ; Isocitrate lyase ; Cloning ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary An Aspergillus nidulans gene library was constructed in a high-frequency transformation vector, pDJB3, based on the Neurospora crassa pyr4 gene. This gene library was used to isolate the structural gene for isocitrate lyase (acuD) by complementation of a deficiency mutation following transformation of A. nidulans. Plasmids rescued in Escherichia coli were able to transform five different A. nidulans acuD mutants. Transformation using plasmids containing the cloned fragment resulted in integration at the acuD locus in six of nine transformants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331649
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  • 23
    Electronic Resource
    Electronic Resource
    Integration of foreign DNA following microinjection of tobacco mesophyll protoplasts (1986)
    Springer
    Molecular genetics and genomics 202 (1986), S. 179-185 
    Details
    ISSN: 1617-4623
    Keywords: Gene transfer ; Plant cell transformation ; Plant tissue culture ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary DNA from a bacterial plasmid containing the T-DNA border sequences of Agrobacterium tumefaciens was transferred into the nucleus or the cytoplasm of tobacco mesophyll protoplasts by microinjection. Following culture in hanging drops, some of these protoplasts produced calli containing the foreign DNA sequences. Evidence for the presence of the injected plasmid DNA in these calli was provided by Southern hybridization analysis. The results demonstrated that random portions of the bacterial plasmid were linked to plant DNA and that integration did not occur at the T-DNA borders present on the injected plasmid. The average number of integrated copies ranged from less than one to 1–2 per tobacco genome. The frequency of integration averaged 14% with intranuclear injections compared to 6% with cytoplasmic injections. With further refinement, the use of microinjection may allow the introduction of many different types of genetic elements into plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331634
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  • 24
    Electronic Resource
    Electronic Resource
    In vivo cloning of DNA into multicopy cosmids by mini-Mu-cosduction (1986)
    Springer
    Molecular genetics and genomics 205 (1986), S. 546-549 
    Details
    ISSN: 1617-4623
    Keywords: Transposition ; Genomic libraries ; Recombinant DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A general in vivo procedure for cloning Escherichia coli genes into cosmids has been developed. The method we describe here uses a deleted Mu phage (a mini-Mu) to transpose E. coli genes into cosmids during mini-Mu replication. The resulting cosmids clones are packaged in-vivo into λ phage particles. Plasmids carrying a particular DNA sequence can be selectively recovered after infection of a new host with the in vivo constructed genomic cosmid library. This system was used succesfully to clone several E. coli genes.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00338096
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