【摘要】 目的构建含6个布鲁氏菌蛋白的原核表达载体,并对其重组蛋白进行诱导表达及纯化。方法以羊种布鲁氏菌16M基因组DNA为模板扩增6个蛋白基因,与载体pET32a(+)连接,转化E.coliBL21(DE3)感受态细胞,IPTG诱导蛋白表达,SDS-PAGE及Westernblot鉴定,采用HisTrapTMHP纯化目的蛋白。结果成功克隆了6个布鲁氏菌蛋白,经对表达条件进行优化,目的蛋白获得大量表达,纯化后纯度达90%以上。结论获得了纯度较高的6个布鲁氏菌蛋白,为进一步研究其免疫原性及保护性奠定基础。更多还原

【Abstract】 The aim of this study is to construct a prokaryotic expression vector for six brucella proteins,as well as express and purify the recombinant protein.Brucella strain 16M genome sequence was extracted and used as template in polymerase chain reactions to amplify six genes,then the PCR product was cloned into the plasmid pET32a(+) and introduced into the competent cells E.coli BL21(DE3).The protein expression was induced by IPTG and examined by SDS-PAGE and Western blot,then purified by HisTrapTM HP.In this study,six brucella proteins were successfully cloned,the expression condition was optimized,the target proteins were strongly expressed,and highly purified protein were obtained after purified.This study lays a foundation for advance study for immunogenicity and protection of the six brucella proteins.更多还原