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Molecular cloning and sequencing of genomic DNA encoding yeast vacuolar carboxypeptidase yscS
Author(s) -
Bordallo Javier,
Bordallo Carmen,
Santiago Gascón,
Suárez-Rendueles Paz
Publication year - 1991
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(91)80546-f
Subject(s) - genomic dna , biology , microbiology and biotechnology , complementation , saccharomyces cerevisiae , molecular cloning , gene , open reading frame , nucleic acid sequence , genetics , genomic library , peptide sequence , biochemistry , mutant
A Saccharomyces cerevisiae genomic DNA encoding vacuolar carboxypeptidase yscS was cloned from a yeast YEp 13 library by complementation of the previously characterized mutation epsl ‐1 [(1981) J. Bacteriol. 147, 418–426], by means of staining carboxypeptidase activity in yeast colonies. The nucleotide sequence of the cloned gene was determined. The open reading frame of CPS1 consists of 576 codons and therefore encodes a protein of 64961 molecular weight. A stretch of 19 residues near the N‐terminus of the deduced polypeptide sequence contains characteristics common to known hydrophobic leader sequences. CPS1 was determined by DNA blot analysis to be a single copy gene located on chromosome X. The cloned fragment was used to identify a 2.1 kb mRNA. A transcriptional activation of CPS1 occurs when cells grow on a substrate of carboxypeptidase yscS as sole nitrogen source.