黑龙江八一农垦大学学报2012，Vol.24Issue(5)：50-54,5.
鼠伤寒沙门氏茵鞭毛蛋白FIiC的原核表达及纯化
Expression and Purification of Fusion Protein of S.typhimurium FIiC
摘要
Abstract
The purpose of this study was to clone, express and identify FliC protein of Salmonella typhimurium in order to provide the foundation for the study of adjuvant effects of FliC. The gene encoding protein FliC was amplified from the genomic DNA of S.typhimurium 8014 by using PCR technique. The amplified product was cloned into pMD18-T. Plasmids containing the right insert were sequenced to confirm its identity,and then cloned into expression vector pQE30 (+). The constructed recombinant plasmid Flic-pQE30 was transformed to E.cali XL1-Blue and induced with IPTG, and the expressed product was identified by SDS-PAGE and Western blot. Sequence analysis showed that the full coding length of Flic was 1 485 bp,which could encode 495 amino acid residues with a molecular mass of 56 kD.The SDS-PAGE electrophoresis analysis showed that the best expression was induced in 5 hours by 37 ~C and 1 mM IFl'G,under which a relative molecular weight of 56 kD recombinant protein FliC was produced. The recombinant FliC showed specific reaction with sera of rats immunized with S.typhimurium by Western blotting,which showed that the recombinant protein has well reactogenicity. The prokaryotic expression vector for fliC gene was constructed successfully,and fusion protein was expressed in E.coli XLl-Blue,and lay the foundation for the study of adjuvant effects of FliC.关键词
鼠伤寒沙门氏菌/鞭毛蛋白/克隆/表达与鉴定Key words
Salmonella typhimurium/Flagellin/cloning/expression and identification分类
农业科技引用本文复制引用
王鹤,梁宏儒,胡旭,赵达,姜东君,尹辉,高佳滨,陈为宏,崔玉东,朱战波..鼠伤寒沙门氏茵鞭毛蛋白FIiC的原核表达及纯化[J].黑龙江八一农垦大学学报,2012,24(5):50-54,5.基金项目
黑龙江省研究生创新科研项目 （）
国家自然科学基金项目（31072120）. （）
