广宿主范围质粒RK2的启动子：对五种革兰氏阴性菌中转录（起始）的分析
Promoters of the broad host range plasmid RK2: analysis of transcription (initiation) in five species of gram-negative bacteria.
作者信息
Greener A, Lehman S M, Helinski D R
机构信息
Department of Biology, University of California, San Diego, La Jolla 92093.
出版信息
Genetics. 1992 Jan;130(1):27-36. doi: 10.1093/genetics/130.1.27.
Abstract
A broad host range cloning vector was constructed, suitable for monitoring promoter activity in diverse Gram-negative bacteria. This vector, derived from plasmid RSF1010, utilized the firefly luciferase gene as the reporter, since the assay for its bioluminescent product is sensitive, and measurements can be made without background from the host. Twelve DNA fragments with promoter activity were obtained from broad host range plasmid RK2 and inserted into the RSF1010 derived vector. The relative luciferase activities were determined for these fragments in five species of Gram-negative bacteria. In addition, four promoters were analyzed by primer extension to locate transcriptional start sites in each host. The results show that several of the promoters vary substantially in relative strengths or utilize different transcriptional start sites in different bacteria. Other promoters exhibited similar activities and identical start sites in the five hosts examined.
摘要
构建了一种广泛宿主范围的克隆载体，适用于监测多种革兰氏阴性菌中的启动子活性。该载体源自质粒RSF1010，利用萤火虫荧光素酶基因作为报告基因，因为其生物发光产物的检测灵敏度高，并且可以在没有宿主背景的情况下进行测量。从广泛宿主范围的质粒RK2中获得了12个具有启动子活性的DNA片段，并将其插入到源自RSF1010的载体中。测定了这些片段在五种革兰氏阴性菌中的相对荧光素酶活性。此外，通过引物延伸分析了四个启动子，以确定每种宿主中的转录起始位点。结果表明，几个启动子的相对强度有很大差异，或者在不同细菌中利用不同的转录起始位点。其他启动子在检测的五种宿主中表现出相似的活性和相同的起始位点。
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本文引用的文献
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