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  • 1
    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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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Molecular genetics of met17 and met25 mutants of Saccharomyces cerevisiae: Intragenic complementation between mutations of a single structural gene (1987)
    Springer
    Molecular genetics and genomics 207 (1987), S. 165-170 
    Details
    ISSN: 1617-4623
    Keywords: Methionine metabolism ; Gene complementation ; MET25 gene ; Sulphydrylase ; Saccharomyces cerevisiae
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We cloned the MET17 gene of Saccharomyces cerevisiae by functional complementation after transformation of a yeast met17 mutant. Restriction mapping and nucleotide sequencing of the MET17 clones revealed that these were from the same genomic region as clones isolated previously and shown to contain the MET25 gene encoding the enzyme O-acetylhomoserine, O-acetylserine sulphydrylase (OAH-OAS sulphydrylase). Transformation studies with MET25 clones showed that the MET17 and MET25 functions were both endoced in a single transcription unit. We conclude that met17 and met25 are both mutations in the structural gene for the OAH-OAS sulphydrylase subunit and that each affects a different fuctional domain of the enzyme allowing subunit complementation in the met17xmet25 diploid. Enzyme assays indicated that the diploid, although not requiring methionine, had a low OAH-OAS sulphydrylase activity (10% of wild type). This is consistent with MET17 and MET25 being the same gene. We found that both met17 and met25 mutants were devoid of 3′ phospho-adenosine 5′ phospho-sulphite (PAPS) reductase activity and that this activity was fully restored in the met17xmet25 diploid. The possible interactions between OAH-OAS sulphydrylase and PAPS reductase are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00331505
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    Paper (German National Licenses)
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  • 3
    Electronic Resource
    Electronic Resource
    The expression of the MET25 gene of Saccharomyces cerevisiae is regulated transcriptionally (1985)
    Springer
    Molecular genetics and genomics 200 (1985), S. 407-414 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The MET25 gene of Saccharomyces cerevisiae was cloned by functional complementation after transformation of a yeast met25 mutant. Subcloning of the DNA fragment bearing MET25 located the gene on a 2.3 kb region. The gene was formally identified by integration at the chromosomal MET25 locus. The cloned MET25 gene was used as a probe to measure the MET25 messenger RNA in a wild-type strain grown under conditions which promoted or failed to promote repression of MET25 expression. It was found that, under repression conditions, MET25 messenger RNA was reduced tenfold when compared with non-repression conditions. This suggests that the expression of MET25 is regulated transcriptionally. The direction of transcription, the size of the transcript and the position of the transcribed part of the gene were determined. Deletion mapping of the regulatory region was carried out. Deleted plasmids were introduced back into yeast cells and tested for their ability to complement met25 mutations and to promote regulation of expression of the MET25 gene by exogenous methionine. By this method the regulatory region was found to be confined to a 130 bp region.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425724
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    Paper (German National Licenses)
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