一株解脂假丝酵母中环酰亚胺抗性基因的克隆与序列分析

Cloning and sequence analysis of a Candida maltosa gene which confers resistance to cycloheximide.

作者信息

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

机构信息

Department of Molecular Biology, Institute of Applied Enzymology, Vilnius, Lithuania.

出版信息

Gene. 1992 Jul 1;116(1):105-8. doi: 10.1016/0378-1119(92)90636-4.

DOI:10.1016/0378-1119(92)90636-4

PMID:1628836

Abstract

A CYHR gene from Candida maltosa, which confers resistance to cycloheximide, was cloned in Saccharomyces cerevisiae. A 2.3-kb DNA fragment carrying this gene was sequenced, and an open reading frame able to encode 553 amino acids (aa) was found in the sequence. Computer searches of the GenBank, EMBL, SWIS-PROT and Gen-Pept databases using the FASTA program failed to detect any proteins with extensive similarities to the deduced aa sequence for CYHR. The cloned gene transforms S. cerevisiae at a frequency similar to auxotrophic markers and can be used as a dominant selectable marker for introducing recombinant plasmids into wild-type strains of S. cerevisiae, as well as for gene disruption experiments.

摘要

克隆了来自麦芽糖假丝酵母的一个赋予对放线菌酮抗性的CYHR基因，该基因在酿酒酵母中表达。对携带此基因的一个2.3kb DNA片段进行了测序，并且在该序列中发现了一个能够编码553个氨基酸（aa）的开放阅读框。使用FASTA程序对GenBank、EMBL、SWIS-PROT和Gen-Pept数据库进行计算机检索，未能检测到与CYHR推导的氨基酸序列有广泛相似性的任何蛋白质。克隆的基因以与营养缺陷型标记相似的频率转化酿酒酵母，并且可作为显性选择标记用于将重组质粒导入酿酒酵母的野生型菌株，以及用于基因破坏实验。