title: Construction of a genomic library of the food spoilage yeast Zygosaccharomyces bailii and isolation of the beta-isopropylmalate dehydrogenase gene (ZbLEU2).
abstract: 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 beta-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 beta-isopropylmalate dehydrogenases from several other yeast species. The sequence of ZbLEU2 has been deposited in the EMBL data library under accession number AJ292544.
authors.affiliation: Centro de Ciências do Ambiente, Departamento de Biologia, Universidad do Minho, Campus de Gualtar, Braga, Portugal.
authors.forename: F
authors.initials: F
authors.lastname: Rodrigues
authors.forename: A M
authors.initials: AM
authors.lastname: Zeeman
authors.forename: C
authors.initials: C
authors.lastname: Alves
authors.forename: M J
authors.initials: MJ
authors.lastname: Sousa
authors.forename: H Y
authors.initials: HY
authors.lastname: Steensma
authors.forename: M
authors.initials: M
authors.lastname: Côrte-Real
authors.forename: C
authors.initials: C
authors.lastname: Leão
mesh_terms: D050539:3-Isopropylmalate Dehydrogenase; D000429:Alcohol Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D003470:Culture Media; D005516:Food Microbiology; D005800:Genes, Fungal; D005816:Genetic Complementation Test; D015698:Genomic Library; D008969:Molecular Sequence Data; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics; D017422:Sequence Analysis, DNA; D020068:Zygosaccharomyces / Q000201:enzymology* / Q000235:genetics
chemical_list: D003470:Culture Media; D000429:Alcohol Oxidoreductases; D050539:3-Isopropylmalate Dehydrogenase