Ramesh G, Gopinathan K P
Department of Microbiology and Cell Biology, Indian Institute of Science, Bangalore.
Indian J Biochem Biophys. 1995 Dec;32(6):361-7.
Restriction fragments of mycobacteriophage I3 DNA capable of initiating transcription have been cloned into a promoter selection vector of Escherichia coli, and selected on the basis of development of resistance to chloramphenicol. The growth pattern of these 'promoter clones' on a concentration gradient of chloramphenicol and the biochemical assays of the chloramphenicol acetyl transferase have permitted the assessment of their relative promoter strengths. DNA sequence analysis revealed significant homology of these promoters to the -35 regions of the mycobacterial--and E. coli promoter consensus, but less so the -10 region. Based on the sequence of phage I3 promoters identified here and the reported sequences of mycobacterial promoters, a promoter consensus for mycobacteria has been generated.
能够启动转录的分枝杆菌噬菌体I3 DNA的限制性片段已被克隆到大肠杆菌的启动子选择载体中，并基于对氯霉素抗性的发展进行筛选。这些“启动子克隆”在氯霉素浓度梯度上的生长模式以及氯霉素乙酰转移酶的生化分析，使得对它们相对启动子强度的评估成为可能。DNA序列分析显示，这些启动子与分枝杆菌和大肠杆菌启动子共有序列的-35区域具有显著同源性，但与-10区域的同源性较低。基于此处鉴定的噬菌体I3启动子序列和已报道的分枝杆菌启动子序列，已生成了分枝杆菌的启动子共有序列。
