Open AccessConstruction of a genomic library of the food spoilage yeast Zygosaccharomyces bailii and isolation of the β‐isopropylmalate dehydrogenase gene ( ZbLEU2 )
Author(s) -
Rodrigues Fernando,
Zeeman AnneMarie,
Alves Cíntia,
Sousa Maria João,
Steensma H.Yde,
CôrteReal Manuela,
Leão Cecília
Publication year - 2001
Publication title -
fems yeast research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.991
H-Index - 92
eISSN - 1567-1364
pISSN - 1567-1356
DOI - 10.1111/j.1567-1364.2001.tb00014.x
Subject(s) - biology , saccharomyces cerevisiae , gene , genomic library , complementation , genetics , yeast , shuttle vector , genome , open reading frame , whole genome sequencing , peptide sequence , mutant , recombinant dna , vector (molecular biology)
A genomic library of the yeast Zygosaccharomyces bailii ISA 1307 was constructed in pRS316, a shuttle vector for Saccharomyces cerevisiae and Escherichia coli . The library has an average insert size of 6 kb and covers the genome more than 20 times assuming a genome size similar to that of S. cerevisiae . This new tool has been successfully used, by us and others, to isolate Z. bailii genes. One example is the β‐isopropylmalate dehydrogenase gene ( ZbLEU2 ) of Z. bailii , which was cloned by complementation of a leu2 mutation in S. cerevisiae . An open reading frame encoding a protein with a molecular mass of 38.7 kDa was found. The nucleotide sequence of ZbLEU2 and the deduced amino acid sequence showed a significant degree of identity to those of β‐isopropylmalate dehydrogenases from several other yeast species. The sequence of ZbLEU2 has been deposited in the EMBL data library under accession number AJ292544.