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白色念珠菌中编码DNA拓扑异构酶I基因的鉴定。

Identification of the gene encoding DNA topoisomerase I from Candida albicans.

作者信息

Taylor A, Giles K, Sarthy A V, McGonigal T, Fostel J

机构信息

Antifungal Biology, Anti-infective Division, Abbott Laboratories, Abbott Park, IL 60064, USA.

出版信息

FEMS Microbiol Lett. 1996 May 1;138(2-3):113-21. doi: 10.1111/j.1574-6968.1996.tb08143.x.

Abstract

A gene encoding a type I topoisomerase (TOP1) was isolated from Candida albicans, sequenced, and expressed in Saccharomyces cerevisiae. The TOP1 gene was identified from a C. albicans genomic library by hybridization with the product of a polymerase chain reaction with degenerate primer sets encoding regions conserved in other TOP1 genes. A clone containing an open reading frame of 2463 bp and predicted to encode a protein of 778 amino acids with sequence similarity to eukaryotic type I topoisomerases was identified. The C. albicans TOP1 gene restored camptothecin sensitivity and increased the topoisomerase activity in S. cerevisiae, indicating that the DNA fragment encodes a functional C. albicans topoisomerase I.

摘要

从白色念珠菌中分离出一个编码I型拓扑异构酶(TOP1)的基因,进行测序,并在酿酒酵母中表达。通过与用编码其他TOP1基因中保守区域的简并引物对进行聚合酶链反应的产物杂交,从白色念珠菌基因组文库中鉴定出TOP1基因。鉴定出一个克隆,其含有一个2463 bp的开放阅读框,预计编码一个778个氨基酸的蛋白质,该蛋白质与真核I型拓扑异构酶具有序列相似性。白色念珠菌TOP1基因恢复了酿酒酵母对喜树碱的敏感性,并增加了其拓扑异构酶活性,表明该DNA片段编码一种功能性白色念珠菌拓扑异构酶I。

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