基于其乳清苷-5'-磷酸脱羧酶基因对纤细内孢霉进行转化。

Transformation of Endomyces fibuliger based on its gene for orotidine-5'-phosphate decarboxylase.

作者信息

Ngan W Y, Nga B H, Pridmore D, Mollet B, Tan H M

机构信息

Department of Microbiology, National University of Singapore, MD4, 5 Science Drive 2, 117597, Singapore, Singapore.

出版信息

Gene. 2000 Aug 22;254(1-2):97-103. doi: 10.1016/s0378-1119(00)00286-9.

DOI:10.1016/s0378-1119(00)00286-9

PMID:10974540

Abstract

Endomyces fibuliger is a yeast used in the production of Chinese rice wine. It secretes enzymes such as glucoamylase, alpha-amylase and acid protease. Very little is known of the genetics of E. fibuliger. In order to develop a transformation system for this yeast, orotidine-5'-phosphate decarboxylase mutant strains were obtained and characterized. Transformation of the E. fibuliger ura3 mutant F1 with an integrative plasmid that carried the wild-type URA3 gene of E. fibuliger gave complementation of this mutation. The E. fibuliger gene encodes the orotidine-5'-phosphate decarboxylase enzyme consisting of 266 amino acid residues with a 69.4% sequence identity with orotidine-5'-phosphate decarboxylase of Saccharomyces cerevisiae. Our finding that E. fibuliger URA3 complements the ura3 mutation in S. cerevisiae confirms that the URA3 gene of E. fibuliger encodes a protein that exerts a similar function.

摘要

扣囊复膜孢酵母是一种用于生产中国米酒的酵母。它能分泌诸如葡糖淀粉酶、α-淀粉酶和酸性蛋白酶等酶。人们对扣囊复膜孢酵母的遗传学了解甚少。为了开发这种酵母的转化系统，获得了乳清苷-5'-磷酸脱羧酶突变菌株并对其进行了表征。用携带扣囊复膜孢酵母野生型URA3基因的整合质粒转化扣囊复膜孢酵母ura3突变体F1，实现了该突变的互补。扣囊复膜孢酵母基因编码由266个氨基酸残基组成的乳清苷-5'-磷酸脱羧酶，与酿酒酵母的乳清苷-5'-磷酸脱羧酶具有69.4%的序列同一性。我们发现扣囊复膜孢酵母URA3能互补酿酒酵母中的ura3突变，这证实了扣囊复膜孢酵母的URA3基因编码一种发挥类似功能的蛋白质。