Shazand K, Tucker J, Grunberg-Manago M, Rabinowitz J C, Leighton T
Institut de Biologie Physico-Chimique, Paris, France.
J Bacteriol. 1993 May;175(10):2880-7. doi: 10.1128/jb.175.10.2880-2887.1993.
We reported previously the cloning and sequence of the Bacillus subtilis infB gene which encodes the essential IF2 factor required for initiation of translation (K. Shazand, J. Tucker, R. Chiang, K. Stansmore, H. U. Sperling-Petersen, M. Grunberg-Manago, J. C. Rabinowitz, and T. Leighton, J. Bacteriol. 172:2675-2687, 1990). The location of the 5' border of the infB operon was investigated by using integrative plasmids carrying various DNA fragments from the region upstream of the infB gene. The lethal effect of disruption of the infB transcriptional unit could be suppressed when the integrated plasmid introduced the spac promoter upstream of the infB operon and transformants were selected in conditions of induction of spac expression. Such an integrated plasmid was used as a starting point to clone the promoter of the infB operon. Primer extension mapping suggests that a single sigma A-type promoter controls transcription of the infB operon. The sequence of a 5,760-bp region encompassing the infB gene was determined. The infB operon is located immediately downstream of the polC gene and comprises seven open reading frames, four of which appear to be the homologs of genes present in the same order in the Escherichia coli infB operon, including nusA. The striking similarity between the E. coli and B. subtilis infB operons suggests that the function of each gene pair is conserved and that the B. subtilis NusA homolog, which is 124 residues shorter than its E. coli counterpart, could play a role similar to its role in E. coli.
我们先前报道了枯草芽孢杆菌infB基因的克隆及序列,该基因编码翻译起始所需的必需IF2因子(K.沙赞德、J.塔克、R.蒋、K.斯坦斯莫尔、H.U.斯珀林 - 彼得森、M.格伦伯格 - 马纳戈、J.C.拉比诺维茨和T.莱顿,《细菌学杂志》172:2675 - 2687,1990年)。通过使用携带来自infB基因上游区域各种DNA片段的整合质粒,研究了infB操纵子5'边界的位置。当整合质粒在infB操纵子上游引入spac启动子并在spac表达诱导条件下选择转化子时,infB转录单元破坏的致死效应可以被抑制。这样的整合质粒被用作克隆infB操纵子启动子的起点。引物延伸图谱表明,一个单一的σA类启动子控制infB操纵子的转录。测定了包含infB基因的5760 bp区域的序列。infB操纵子位于polC基因的紧邻下游,由七个开放阅读框组成,其中四个似乎是大肠杆菌infB操纵子中按相同顺序存在的基因的同源物,包括nusA。大肠杆菌和枯草芽孢杆菌infB操纵子之间的显著相似性表明,每对基因的功能是保守的,并且枯草芽孢杆菌的NusA同源物比其大肠杆菌对应物短124个残基,可能发挥与其在大肠杆菌中类似的作用。