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乳酸发酵短杆菌ftsZ基因的鉴定、特性分析及染色体组织

Identification, characterization, and chromosomal organization of the ftsZ gene from Brevibacterium lactofermentum.

作者信息

Honrubia M P, Fernández F J, Gil J A

机构信息

Departamento de Ecologia, Genética y Microbiología, Facultad de Biología, Universidad de León, Spain.

出版信息

Mol Gen Genet. 1998 Jul;259(1):97-104. doi: 10.1007/s004380050793.

Abstract

The ftsZ gene was cloned from the chromosomal DNA of Brevibacterium lactofermentum by the polymerase chain reaction (PCR) using two oligonucleotides designed from two conserved regions found in most of the previously cloned and sequenced ftsZ genes from other microorganisms. ftsZ is a single-copy gene in corynebacteria and is located downstream from ftsQ and murC, indicating linkage between genes involved in peptidoglycan synthesis (mur genes) and genes involved in cell division (fts genes). The organisation of the cluster is similar to that in Streptomyces and different from those of Escherichia coli or Bacillus subtilis because ftsA is not located upstream of ftsZ. The gene was expressed in E. coli using the T7 expression system; the calculated molecular weight of the expressed protein was 50 kDa. Expression of the B. lactofermentum ftsZ gene in E. coli inhibited cell division and led to filamentation. The ftsZ gene of this organism does not complement ftsZ mutations or deletions in E. coli, when cloned on low or high-copy-number vectors.

摘要

通过聚合酶链反应(PCR),使用从其他微生物中先前克隆和测序的大多数ftsZ基因中发现的两个保守区域设计的两个寡核苷酸,从乳酸发酵短杆菌的染色体DNA中克隆ftsZ基因。ftsZ是棒状杆菌中的单拷贝基因,位于ftsQ和murC的下游,表明参与肽聚糖合成的基因(mur基因)和参与细胞分裂的基因(fts基因)之间存在联系。该基因簇的组织方式与链霉菌中的相似,与大肠杆菌或枯草芽孢杆菌的不同,因为ftsA不在ftsZ的上游。使用T7表达系统在大肠杆菌中表达该基因;表达蛋白的计算分子量为50 kDa。乳酸发酵短杆菌ftsZ基因在大肠杆菌中的表达抑制了细胞分裂并导致丝状化。当克隆到低拷贝数或高拷贝数载体上时,该生物体的ftsZ基因不能互补大肠杆菌中的ftsZ突变或缺失。

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