柯萨奇B1病毒VP1基因在大肠杆菌中的表达

Expression of VP1 gene of Coxsackievirus B1 in E coli

目的 在大肠杆菌中表达柯萨奇 B1(CVB1)病毒 VP1蛋白。方法 用限制性酶切方法将 CVB1VP1基因从 p MD18-T-VP1上切下 ,连接至 p QE3 0构建原核表达系统 p QE3 0 -VP1。经酶切和 PCR验证后转化至大肠杆菌 BL2 1(DE3 ) ,用 IPTG诱导目的基因表达。SDS-聚丙烯酰胺凝胶电泳和 Western-blot检测 VP1蛋白的表达。结果 酶切、PCR证明构建的原核表达系统携带方向正确的 CVB1VP1基因。SDS-PAGE和 Western-blot实验均可于 3 3 .0 k Da处见到目的条带。结论

Objective To express the VP1 gene of Coxsackievirus B1 in E coli.Methods CVB1 VP1 was got from pMD18 T VP1 with restrictive endonuclease and cloned into pQE30 to construct the prokaryotic expression system pQE30 VP1. Confirmed with restriction analysis and PCR, pQE30 VP1 was transformed into E coli BL21(DE3) and induced by IPTG. SDS PAGE and Western blot were used to detect the VP1 protein.Results Restriction analysis and PCR showed that CVB1 VP1 was cloned into pQE30 in correct direction. SDS PAGE and Western blot showed the 33.0 kDa protein band of VP1 clearly.Conclusions The prokaryotic expression of CVB1 VP1 may provide the basis for the development of a new diagnostic reagent.