口蹄疫病毒3A基因在大肠杆菌中的高效表达

Study on the Expression of 3A Gene of Foot-and-mouth Disease in E.coli

将口蹄疫病毒 (Foot and MouthDiseaseVirus,FMDV)的 3A基因克隆到线形化的原核表达载体pProEX HTb中 ,转化大肠杆菌BL2 1和DH5α ,经氨苄抗性筛选得到阳性克隆 ,IPTG诱导表达。SDS PAGE和Westbolt结果证实大肠杆菌菌体不可溶性蛋白中富含 3A蛋白 ,说明3A蛋白在表达产物中以包涵体的形式存在 ,所表达的蛋白含量占菌体蛋白的 2 9 2 %。

A gene of foot-and-mouth diease viruse(FMDV) was cloned into linearizied pProEX-HTb prokaryotic expresion vector. The vector was transformed into E.coli BL21, the transformants were screened by Amp++ and were induced to express with IPTG. The reults of SDS-PAGE and Western blot demonstrated that the insoulable component of the induced E.coli culture contained protain 3A. The results of the study indicated that protain 3A existed in the form of inclusion body. The content of the expressed protain in the induced bacteria protain was 29.2%.