重组大肠杆菌表达水稻白叶枯病菌FtsZ蛋白

摘要/Abstract

摘要： 旨在通过现代分子生物学技术制备水稻白叶枯病菌FtsZ蛋白。以水稻白叶枯病菌总DNA为模板,采用巢式PCR方法扩增获得水稻白叶枯病菌ftsZ基因,构建ftsZ基因的表达载体pET-22b-ftsZ,转化表达宿主E. coli BL21后,经PCR、Nde I/Xho I双酶切及测序鉴定、阳性克隆子经IPTG诱导表达,融合蛋白经镍柱纯化后,通过SDS-PAGE和Western blotting分析鉴定。结果显示,水稻白叶枯病菌ftsZ基因的重组表达载体构建成功,且阳性克隆子在IPTG的诱导下表达了FtsZ-6×His融合蛋白,并通过镍柱纯化获得了电泳纯的FtsZ-6×His融合蛋白。

关键词: FtsZ蛋白, 水稻白叶枯病菌, 大肠杆菌, 基因重组

Abstract: It was to prepare FtsZ protein using techniques of modern molecular biology. The ftsZ gene was amplified from Xanthomonas oryzae by nested PCR, and recombinant plasmid pET-22b-ftsZ was constructed and transformed to E.coli BL21. The clony fragment was identificatified by PCR screening, Nde I/Xho I digestion and DNA sequencing, the positive clones were induced by IPTG for expression；the fusion protein was purified through Ni-NTA Resin, and identified by SDS-PAGE and Western blotting. Results showed that the recombinant plasmid pET-22b-ftsZ was constructed successfully, the FtsZ-6×His fusion protein was expressed in recombined E. coli BL21 induced by IPTG, and purified through Ni-NTA Resin by electrophoretic purity.

Key words: FtsZ protein, Xanthomonas oryzae , Escherichia coli , Genetic recombination

陈洋,黄运红,李素珍,龙中儿. 重组大肠杆菌表达水稻白叶枯病菌FtsZ蛋白[J]. 生物技术通报, 2014, 0(11): 174-178.

Chen Yang, Huang ,Yunhong, Li Suzhen, Long Zhonger. Expression of FtsZ Protein from Xanthomonas oryzae in Escherichia coli[J]. Biotechnology Bulletin, 2014, 0(11): 174-178.

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