摘要
从大肠杆菌K88(LT+,ST+)菌株中扩增热敏肠毒素B亚基基因(ltb),得到454 bp片段,克隆至pMD18-T载体后,与pET32a(+)定向连接,并转化入大肠杆菌BL21中,用IPTG 30℃诱导表达重组LTB蛋白。SDS-PAGE显示,重组蛋白分子量约为34 kDa,超声波裂解后,表达产物主要以包涵体形式存在。经鉴定,纯化复性后的LTB蛋白保留部分与GM1结合的生物学活性。
The gene of Escherichia coli heat-labile enterotoxin subunit B was cloned into plasmid pET32a vector and transformed into E.coli BL21.The recombinant protein was expressed after being induced by IPTG at 30℃.The SDS-PAGE indicated that the molecular weight of recombinant LTB was about 34 kDa.Most of the recombinant protein existed in inclusion body after ultrasonication,and part of protein regained bio-activity of binding with GM1 after renaturation.
出处
《中国动物传染病学报》
CAS
2011年第2期31-36,共6页
Chinese Journal of Animal Infectious Diseases
