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  • Artikel  (378)
  • Female  (358)
  • Recombinant DNA
  • 1985-1989  (378)
  • Biologie  (378)
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  • Architektur, Bauingenieurwesen, Vermessung
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  • Artikel  (378)
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  • 1
    Digitale Medien
    Digitale Medien
    The isolation of specific genes from the basidiomycete Schizophyllum commune (1987)
    Springer
    Current genetics 12 (1987), S. 547-554 
    Details
    ISSN: 1432-0983
    Schlagwort(e): Schizophyllum commune ; Transformation ; Gene isolation ; Basidiomycetes ; Recombinant DNA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00419565
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  • 2
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): Recombinant DNA ; Cross hybridization ; 2D-electrophoresis ; Hybrid selection translation ; Transformation
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00418408
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  • 3
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): Mitochondrial DNA ; Population heterogeneity ; Molecular evolution ; Recombinant DNA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00389428
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  • 4
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): Fermentation ; Recombinant DNA ; Hepatitis B surface antigen
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Werkstoffwissenschaften, Fertigungsverfahren, Fertigung
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF01569510
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  • 5
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): metF ; 5,10-methylenetetrahydrofolate reductase ; Recombinant DNA ; Nucleotide sequence
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00334692
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  • 6
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): metC ; Cystathionine-β-lyase ; Nucleotide sequence ; Recombinant DNA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00332246
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  • 7
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): Recombinant DNA ; Methionine metabolism ; Negative control ; Regulatory regions
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00325699
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  • 8
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): Recombinant DNA ; Helix-turn-helix motifs ; In vitro transcription-translation ; Phage immunity ; Exonuclease III deletions
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00339591
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  • 9
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): Aspergillus ; Isocitrate lyase ; Cloning ; Recombinant DNA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00331649
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  • 10
    Digitale Medien
    Digitale Medien
    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
    Schlagwort(e): Gene transfer ; Plant cell transformation ; Plant tissue culture ; Recombinant DNA
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie
    Notizen: 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.
    Materialart: Digitale Medien
    URL: http://dx.doi.org/10.1007/BF00331634
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