赋予酿酒酵母对甲醛抗性的麦芽糖假丝酵母基因的克隆与分析。

Cloning and analysis of a Candida maltosa gene which confers resistance to formaldehyde in Saccharomyces cerevisiae.

作者信息

Sasnauskas K, Jomantiene R, Januska A, Lebediene E, Lebedys J, Janulaitis A

机构信息

Department of Molecular Biology, Institute of Biotechnology, Vilnius, Lithuania.

出版信息

Gene. 1992 Dec 1;122(1):207-11. doi: 10.1016/0378-1119(92)90052-q.

DOI:10.1016/0378-1119(92)90052-q

PMID:1339376

Abstract

A gene (FDH1) of Candida maltosa which confers resistance to formaldehyde in Saccharomyces cerevisiae was cloned and its nucleotide sequence determined. The gene has a single intron which possesses the highly conserved splicing signals found in S. cerevisiae introns. We demonstrated that processing of the pre-mRNA of the cloned gene occurred identically in both S. cerevisiae and C. maltosa. The predicted amino acid sequence from the cloned gene showed 65.5% identity to human alcohol dehydrogenase (ADH) class III and 23.9% identity to S. cerevisiae ADH1. The most probable mechanism of resistance to formaldehyde is thought to be the glutathione-dependent oxidation of formaldehyde which is characteristic for ADH class III. The cloned FDH1 gene was successfully employed as a dominant selectable marker in the transformation of S. cerevisiae.

摘要

克隆了麦芽糖假丝酵母中赋予酿酒酵母对甲醛抗性的一个基因（FDH1），并测定了其核苷酸序列。该基因有一个单一内含子，具有在酿酒酵母内含子中发现的高度保守的剪接信号。我们证明，克隆基因的前体mRNA在酿酒酵母和麦芽糖假丝酵母中的加工方式完全相同。从克隆基因预测的氨基酸序列与人类Ⅲ类乙醇脱氢酶（ADH）有65.5%的同一性，与酿酒酵母ADH1有23.9%的同一性。对甲醛抗性最可能的机制被认为是Ⅲ类ADH特有的依赖谷胱甘肽的甲醛氧化。克隆的FDH1基因在酿酒酵母的转化中成功用作显性选择标记。