麦芽糖假丝酵母自主复制序列的分子克隆与分析

Molecular cloning and analysis of autonomous replicating sequence of Candida maltosa.

作者信息

Sasnauskas K, Jomantienè R, Lebedienè E, Lebedys J, Januska A, Janulaitis A

机构信息

Institute of Applied Enzymology, Vilnius, Lithuanian Republic.

出版信息

Yeast. 1992 Apr;8(4):253-9. doi: 10.1002/yea.320080403.

DOI:10.1002/yea.320080403

PMID:1514324

Abstract

A Candida maltosa chromosomal DNA fragment which confers high frequency transformation of C. maltosa and autonomous replication of recombinant plasmids was cloned and sequenced. Analysis of the nucleotide sequence of the cloned DNA revealed a sequence homologous for C. maltosa autonomously replicating sequence (ARS) elements. Vector pRJ1 for C. maltosa was constructed, which contained a 1.3 kb ARS sequence, pICEM-19H and the ADE1 gene of C. maltosa. Southern blot analysis suggested that the copy number of pRJ1 in C. maltosa was approximately 20 per genome. The sequence analysis also revealed an open reading frame, encoding a polypeptide with high homology (70%) to the RS15 protein of Brugia pagangi. This open reading frame has an intron with canonical sites for correct splicing in Saccharomyces cerevisiae.

摘要

克隆并测序了一个赋予麦芽糖假丝酵母高频转化和重组质粒自主复制能力的麦芽糖假丝酵母染色体DNA片段。对克隆DNA的核苷酸序列分析揭示了与麦芽糖假丝酵母自主复制序列（ARS）元件同源的序列。构建了用于麦芽糖假丝酵母的载体pRJ1，其包含1.3 kb的ARS序列、pICEM - 19H和麦芽糖假丝酵母的ADE1基因。Southern印迹分析表明，麦芽糖假丝酵母中pRJ1的拷贝数约为每个基因组20个。序列分析还揭示了一个开放阅读框，其编码的多肽与马来布鲁线虫的RS15蛋白具有高度同源性（70%）。这个开放阅读框有一个内含子，在酿酒酵母中有用于正确剪接的典型位点。