Zakrzewska-Czerwińska J, Nardmann J, Schrempf H
Department of Microbiology, Institute of Immunology and Experimental Therapy, Wrocław, Poland.
Mol Gen Genet. 1994 Feb;242(4):440-7. doi: 10.1007/BF00281794.
The dnaA gene of Streptomyces lividans was cloned using the Escherichia coli medium-copy-number vector pSU18 and E. coli strain TC1963, which can by-pass the requirement for the DnaA protein. Its regulatory region was subcloned in the Streptomyces probe vector pIJ4083. Primer extension and S1 mapping studies allowed the identification of a class I Streptomyces promotor (P2). An additional, previously unknown promoter type (P1) was found by S1 mapping. The presence of two DnaA box motifs between P1 and P2 suggests that the transcriptions of the S. lividans dnaA gene is autoregulated by its gene product. It was shown that the transcription of the dnaA gene is significantly induced by mitomycin C, an agent known to inhibit DNA replication. The data suggest that, as in E. coli, one of the regulatory mechanisms governing the transcription of the dnaA gene in S. lividans is probably related to the SOS response network.
使用大肠杆菌中拷贝数载体pSU18和能够绕过对DnaA蛋白需求的大肠杆菌菌株TC1963,克隆了变铅青链霉菌的dnaA基因。其调控区被亚克隆到链霉菌探针载体pIJ4083中。引物延伸和S1作图研究确定了一个I类链霉菌启动子(P2)。通过S1作图发现了另一种以前未知的启动子类型(P1)。P1和P2之间存在两个DnaA框基序,这表明变铅青链霉菌dnaA基因的转录受其基因产物的自动调控。结果表明,丝裂霉素C(一种已知能抑制DNA复制的试剂)可显著诱导dnaA基因的转录。数据表明,与大肠杆菌一样,变铅青链霉菌中调控dnaA基因转录的调控机制之一可能与SOS反应网络有关。