Construction of a genomic library of the food spoilage yeast Zygosaccharomyces bailii and isolation of the β-isopropylmalate dehydrogenase gene (ZbLEU2)

被引：12

作者：

Rodrigues, Fernando

Zeeman, Anne-Marie

Alves, Cintia

Sousa, Maria Joao

Steensma, H. Yde

Corte-Real, Manuela

Leao, Cecilia

机构：

[1] Univ Minho, Ctr Ciencias Ambiente, P-4710057 Braga, Portugal

[2] Leiden Univ, Inst Mol Plant Sci, Clusius Lab, NL-2333 AL Leiden, Netherlands

[3] Delft Univ Technol, Kluyver Inst Biotechnol, NL-2628 BC Delft, Netherlands

[4] IPATIMUP, P-4200 Oporto, Portugal

来源：

FEMS YEAST RESEARCH | 2001年 / 1卷 / 01期

关键词：

genomic library; food spoilage yeast; cloning; LEU2; Zygosaccharomyces bailii;

D O I：

10.1016/S1567-1356(01)00006-X

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

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.

引用

页码：67 / 71

页数：5