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[支原体关键分裂蛋白FtsZ基因在大肠杆菌中的克隆与表达]

[Cloning and expression of the gene for the mycoplasma key division protein FtsZ in Escherichia coli].

作者信息

Kukekova A V, Malinin A Iu, Vonskiĭ M A, Borkhsenius S N

机构信息

Institute of Cytology, Russian Academy of Sciences, St. Petersburg, Russia.

出版信息

Genetika. 1999 Mar;35(2):314-21.

Abstract

Gene ftsZ responsible for division of bacterial cells was revealed in most prokaryote groups. A 520-bp fragment of the ftsZ gene was amplified on the template of A. laidlawii DNA using degenerate primers. This fragment was sequenced and served as a hybridization probe for cloning of the full-sized copy of the A. laidlawii ftsZ gene. The amplified fragment was cloned in a pGEX3X vector and expressed in E. coli cells. Polyclonal antibodies derived from the chimeric polypeptide containing a fragment of A. laidlawii FtsZ protein interacted only with the A. laidlawii protein with molecular mass of 40 kDa. Comparison of nucleotide sequences of the ftsZ-gene region of A. laidlawii and other bacterial species showed that they were highly homologous in A. laidlawii, E. coli, and Bac. subtilis, while low homology was revealed between the A. laidlawii sequence and those of the members of the genus Mycoplasma. Analysis of the ftsZ-gene nucleotide sequences is suggested as a means to study the evolutionary relatedness of prokaryotes.

摘要

在大多数原核生物群体中都发现了负责细菌细胞分裂的基因ftsZ。使用简并引物在莱氏无胆甾原体DNA模板上扩增出ftsZ基因的一个520碱基对片段。对该片段进行测序,并将其用作克隆莱氏无胆甾原体ftsZ基因全长拷贝的杂交探针。扩增片段克隆到pGEX3X载体中并在大肠杆菌细胞中表达。源自含有莱氏无胆甾原体FtsZ蛋白片段的嵌合多肽的多克隆抗体仅与分子量为40 kDa的莱氏无胆甾原体蛋白相互作用。莱氏无胆甾原体与其他细菌物种的ftsZ基因区域核苷酸序列比较表明,它们在莱氏无胆甾原体、大肠杆菌和枯草芽孢杆菌中高度同源,而莱氏无胆甾原体序列与支原体属成员的序列之间同源性较低。建议对ftsZ基因核苷酸序列进行分析,作为研究原核生物进化相关性的一种手段。

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