O型口蹄疫病毒VP1嵌合基因的构建及原核表达

首先合成我国O型口蹄疫病毒2个疫苗毒株VP1基因的3个抗原袁位，与另外扩增的流行毒株VP1基因末端273bp片段相连,构建出O型VP1嵌合基因片段（VP1O）。然后，将VPIO基因连接到原核表达载体pET28a上，构建了重组表达质粒pET28a-VP1O，转化大肠杆菌BL21（DE3）进行诱导表达。SDS-PAGE电泳表明。VP1O基因在大肠杆菌中获得表达。Western blotting检测证实表达的VP1O蛋白具有良好的生物学活性。对表达蛋白通过包涵体洗涤的方法进行初步纯化，获得了较高纯度的VP1O蛋白。

Tandem and Prokaryotic Expression of VP10 Gene of Type O FMDV

Three epitopes of two Chinese vaccine strains of type O FMDV were synthesised.And a 273 bp fragment of the VP1 C terminal of another field isolate was cloned into pMD18-T vector by PCR.The three epitopes and fragment of PCR were ligated by cutting sites,and VP1O fragment was constructed.Then the VP1O gene was cloned into prokaryotic expression vectors pET28a,and the recombinant plasmids of pET28a-VP1O was constructed.The plasmids were transformed to BL21(DE3) competent cell.The expression and the immunogenicity of the target gene was detected and proved through SDS-PAGE and Western-blotting.Finally,The protein was purified by the washing of inclusion bodies.