Construction of a genomic library of the food spoilage yeast Zygosaccharomyces bailii and isolation of the beta-isopropylmalate dehydrogenase gene (ZbLEU2).
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.
Rodrigues
F
F
Centro de Ciências do Ambiente, Departamento de Biologia, Universidad do Minho, Campus de Gualtar, Braga, Portugal.
Zeeman
A M
AM
Alves
C
C
Sousa
M J
MJ
Steensma
H Y
HY
Côrte-Real
M
M
Leão
C
C
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
D003470:Culture Media; D000429:Alcohol Oxidoreductases; D050539:3-Isopropylmalate Dehydrogenase