Position: Home > Articles > Expression of Fts Z Protein from Xanthomonas oryzae in Escherichia coli
Biotechnology Bulletin
2014
(11)
174-178
重组大肠杆菌表达水稻白叶枯病菌FtsZ蛋白
作 者：
陈洋;黄运红;李素珍;龙中儿
单 位：
江西师范大学生命科学学院
关键词：
FtsZ蛋白;水稻白叶枯病菌;大肠杆菌;基因重组
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fts Z基因,构建fts Z基因的表达载体p ET-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Fts Z-6×His融合蛋白,并通过镍柱纯化获得了电泳纯的Fts Z-6×His融合蛋白。
译 名：
Expression of Fts Z Protein from Xanthomonas oryzae in Escherichia coli
作 者：
Chen Yang;Huang Yunhong;Li Suzhen;Long Zhonger;College of Life Science,Jiangxi Normal University;
关键词：
FtsZ protein;;Xanthomonas oryzae;;Escherichia coli;;Genetic recombination
摘 要：
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 p ET-22b-fts Z 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 p ET-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.
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