Black C G, Fyfe J A, Davies J K
Department of Microbiology, Monash University, Clayton, Victoria, Australia.
J Bacteriol. 1995 Apr;177(8):1952-8. doi: 10.1128/jb.177.8.1952-1958.1995.
A recombinant plasmid capable of restoring UV resistance to an Escherichia coli uvrB mutant was isolated from a genomic library of Neisseria gonorrhoeae. Sequence analysis revealed an open reading frame whose deduced amino acid sequence displayed significant similarity to those of the UvrB proteins of E. coli, Micrococcus luteus, and Streptococcus pneumoniae. A gonococcal uvrB mutant was constructed and found to be extremely sensitive to UV radiation. Transcriptional fusions between portions of the gonococcal uvrB upstream region and a reporter gene were used to localize promoter activity, and the transcriptional start point of the gonococcal uvrB gene was mapped in E. coli by primer extension. A corresponding sigma 70 promoter was identified within a copy of the 26-bp neisserial repeat, and this identification provided the first evidence of a promoter associated with this repetitive element in N. gonorrhoeae.
从淋病奈瑟菌基因组文库中分离出一种能够恢复大肠杆菌uvrB突变体紫外线抗性的重组质粒。序列分析显示一个开放阅读框,其推导的氨基酸序列与大肠杆菌、藤黄微球菌和肺炎链球菌的UvrB蛋白具有显著相似性。构建了淋病奈瑟菌uvrB突变体,发现其对紫外线辐射极为敏感。利用淋病奈瑟菌uvrB上游区域部分与报告基因之间的转录融合来定位启动子活性,并通过引物延伸在大肠杆菌中绘制了淋病奈瑟菌uvrB基因的转录起始点。在26 bp淋病奈瑟菌重复序列的一个拷贝中鉴定出一个相应的σ70启动子,这一鉴定为淋病奈瑟菌中与该重复元件相关的启动子提供了首个证据。