重组大肠杆菌表达水稻白叶枯病菌FtsZ蛋白  

Expression of Fts Z Protein from Xanthomonas oryzae in Escherichia coli

作　　者：陈洋[1] 黄运红[1] 李素珍[1] 龙中儿[1] 

机构地区：[1]江西师范大学生命科学学院,南昌330022

出　　处：《生物技术通报》2014年第11期174-178,共5页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31160029;31360018);江西省自然科学基金项目(20122BAB204008;20132BAB204007)

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。It was to prepare FtsZ protein using techniques of modern molecular biology. TheftsZ gene was amplified fromXanthomonas oryzae by nested PCR, and recombinant plasmid pET-22b-ftsZ was constructed and transformed toE.coli BL21. The clony fragment was identificatified by PCR screening,NdeI/XhoI 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＆#215;His fusion protein was expressed in recombinedE. coli BL21 induced by IPTG, and purified through Ni-NTA Resin by electrophoretic purity.

关 键 词：FtsZ蛋白 水稻白叶枯病菌 大肠杆菌 基因重组 

分 类 号：S435.111.47[农业科学—农业昆虫与害虫防治]