郑州大学学报（医学版）Issue(4)：505-507,508,4.DOI:10.13705/j.issn.1671-6825.2014.04.017
T7噬菌体尾蛋白 P11的表达、纯化及鉴定
Expression,purification and identification of T7 bacteriophage tail protein P11
赵翠娇 1宁保安 2范献军 2李桂敏 2高志贤1
作者信息
- 1. 郑州大学公共卫生学院营养与食品卫生学教研室郑州450001
- 2. 军事医学科学院卫生学环境医学研究所天津300050
- 折叠
摘要
Abstract
To express,purify and identify capsid tail protein P11 of T7 bacteriophage.Methods:Gene of pro-tein P11 was amplified by PCR , digested with restriction endonuclease EcoRⅠand XhoⅠand then inserted into the expres-sion vector pTIG-TRX.The recombinant vector was transformed to E.coli BL21(DE3).After IPTG induction,the recombi-nant protein was purified by metal chelate chromatography .Its molecular weight was identified by SDS-PAGE and its activity was analysed by Western blot .The recombinant protein P 11 was used to immune rabbit .Finally,the rabbit polycolonal anti-body against protein P11 were obtained.Results:Gene of protein P11 was amplified by PCR.The recombinant protein P11 with 6 ×His tag has been successfully expressed after induction .SDS-PAGE analysis suggested that the molecular weight of the expressed protein was approximately 25 000 .Western blot analysis suggested that the polycolonal antibodidy could rec-ognize T7 phage and P11.Conclusion:We have purified P11 fusion protein and prepared the rabbit polycolonal antibody a-gainst protein P11 successfully ,which lays foundation for the application of T 7 phage display .关键词
T7噬菌体/P11蛋白/多克隆抗体Key words
T7 bacteriophage/P11 protein/polycolonal antibody分类
医药卫生引用本文复制引用
赵翠娇,宁保安,范献军,李桂敏,高志贤..T7噬菌体尾蛋白 P11的表达、纯化及鉴定[J].郑州大学学报（医学版）,2014,(4):505-507,508,4.
