动物医学进展Issue(5)：12-16,5.
产肠毒素大肠埃希菌 K99菌毛基因的克隆与原核表达
Cloning and Prokaryotic Expression of Enterotoxigenic E.coli K99 Fimbria Gene
摘要
Abstract
Enterotoxigenic E.coli (ETEC)K99 complete coding frame was amplified from ETEC strain C83912 DNA with a pair of specific primers based on K99 gene.The PCR product was cloned into pMD19-T vector.Posi-tive recombinant plasmid pMD19-T-K99 was identified correctly by PCR,double-enzyme digestion and sequencing. The obtained double-enzyme digested K99 product was cloned into pET-32a(+)prokaryotic expression vector forming pET-32a(+)-K99 plasmid for expressing in BL21 (DE3).Recombinant K99 was highly expressed.West-ern-blot analysis confirmed that the recombinant K99 protein reacted specifically with K99 mono-factor seraum and immunized rabbits obtained high titer antibodies,showing its good antigenicity.The constructed ETEC K99 pro-karyotic expression plasmid and antigenicity of the recombinant K99 protein laid the foundation for K99 subunit vaccine research and K99 specific antibody preparation.关键词
产肠毒素大肠埃希菌/K99 菌毛/克隆/原核表达/抗原性Key words
Enterotoxigenic E.coli/fimbia K99/cloning/prokaryotic expression/antigenicity分类
农业科技引用本文复制引用
姜宣鹏,张焕容..产肠毒素大肠埃希菌 K99菌毛基因的克隆与原核表达[J].动物医学进展,2014,(5):12-16,5.基金项目
西南民族大学研究生创新课题（CX2013SZ75）；国家“十二五”科技支撑计划课题 （）
