军事医学Issue(10)：799-802,4.DOI:10.7644/j.issn.1674-9960.2014.10.013
肠出血性大肠杆菌O157∶H7毒力蛋白NleB1的表达、纯化及多克隆抗体的制备与鉴定
Prokaryotic expression, purification, identification, and polyclonal antibody prepa-ration of enterohemorrhagic Escherichia coli effector NleB1
陈欣欣 1廖翔 2宋婷 1周围 1戴红梅 1岳俊杰 1王羽 1王玉瑞 1梁龙1
作者信息
- 1. 军事医学科学院生物工程研究所，北京 100071
- 2. 安徽大学，合肥 230601
- 折叠
摘要
Abstract
Objective To construct a prokaryotic plasmid expressing the recombinant protein of enterohemorrhagic Escherichia coli(EHEC) effector NleB1 and to prepare the polyclonal antibody of mouse anti-NleB1.Methods The nleB1 (990 bp) gene was amplified from the genome EHEC O157∶H7 and cloned into the expression plasmid pET24a to construct the recombinant plasmid pET24a-nleB1 that was transformed into E.coli BL21(DE3).After induction with isopropylthio-gelactoside( IPTG) , the His-tag fusion proteins were purified by Ni+affinity chromatography and gel slices.The polyclonal antibody was prepared by immunizing BALB/c mice with purified recombinant proteins and analyzed by Western blotting and ELISA.Results The pET24a-nleB1 recombinant plasmid was successfully constructed, the fusion protein was ex-pressed and purified,and the polyclonal antibody was obtained by immunizing mice with purified fusion protein.Western blotting and ELISA staining demonstrated that the polyclonal antibody was successfully obtained.Conclusion The prepara-tion of the polyclonal antibody against EHEC O157∶H7 NleB1 will be of help for further studies on the function of NleB1 protein.关键词
肠出血性大肠杆菌/NleB1/原核表达/蛋白纯化/多克隆抗体Key words
enterohemorrhagic Escherichia coli/NleB1/prokaryotic expression/protein purification/polyclonal antibody分类
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陈欣欣,廖翔,宋婷,周围,戴红梅,岳俊杰,王羽,王玉瑞,梁龙..肠出血性大肠杆菌O157∶H7毒力蛋白NleB1的表达、纯化及多克隆抗体的制备与鉴定[J].军事医学,2014,(10):799-802,4.
