Tozawa Y, Yoshikawa H, Kawamura F, Itaya M, Takahashi H
Institute of Applied Microbiology, University of Tokyo, Japan.
Biosci Biotechnol Biochem. 1992 Dec;56(12):1995-2002. doi: 10.1271/bbb.56.1995.
The complete set of groES and groEL gene homologues from Bacillus subtilis Marburg 168 was identified, cloned, and characterized. The nucleotide sequence indicated the presence of two open reading frames corresponding to the groES and groEL genes. The presumptive GroES and GroEL proteins were calculated to be polypeptides of 10,175 and 57,175 Da, respectively, and showed extensive sequence similarities with the known GroES and GroEL proteins of Escherichia coli and Mycobacterium tuberculosis. A heat-inducible transcript initiated upstream of the groES coding region was identified by primer-extension analysis of in vivo transcripts, indicating that the two genes consist of an operon. At least six heat-shock inducible proteins were identified in the cell extract of heat treated B. subtilis. Two proteins of 10 and 60 kDa overproduced in B. subtilis cells carrying a multi-copy groES and groEL plasmid were demonstrated to correspond to two out of the six heat-shock inducible proteins. The groES and groEL genes of B. subtilis were physically mapped on the 60 degrees region of a 360 degrees map and genetically mapped at the position of 40% linkage with the purB locus using PBS1 transduction of the groEL genes tagged with a chloramphenicol resistance (chlr) marker.
对枯草芽孢杆菌马伯格168株的groES和groEL基因同源物全套进行了鉴定、克隆和表征。核苷酸序列表明存在两个与groES和groEL基因相对应的开放阅读框。推测的GroES和GroEL蛋白经计算分别为10,175 Da和57,175 Da的多肽，并与大肠杆菌和结核分枝杆菌已知的GroES和GroEL蛋白表现出广泛的序列相似性。通过对体内转录本进行引物延伸分析，鉴定出一个在groES编码区上游起始的热诱导转录本，表明这两个基因组成一个操纵子。在热处理的枯草芽孢杆菌细胞提取物中鉴定出至少六种热休克诱导蛋白。在携带多拷贝groES和groEL质粒的枯草芽孢杆菌细胞中过量产生的两种10 kDa和60 kDa蛋白被证明对应于六种热休克诱导蛋白中的两种。枯草芽孢杆菌的groES和groEL基因在360°图谱的60°区域进行了物理定位，并使用带有氯霉素抗性（chlr）标记的groEL基因的PBS1转导在与purB基因座40%连锁的位置进行了遗传定位。
