JC病毒蛋白VP3原核表达载体的构建及诱导表达

Construction and proeukaryotic expression of a novel VP3 protein of JC virus

目的构建JC病毒蛋白VP3原核表达载体,并观察其表达情况。方法通过聚合酶链式反应(PCR)获得VP3基因,将其连接到pGEM-T载体,测序正确后插入至原核表达载体pET-32a(+)中,转化BL21大肠杆菌,IPTG诱导,并通过十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、Western blot免疫印迹分析鉴定融合蛋白的表达情况。结果扩增获得VP3基因片段,成功构建了大肠杆菌原核表达JC病毒VP3载体。经IPTG诱导,得到了分子质量为59 ku的目的蛋白,Western blot证实其具有良好的抗原性。结论本研究成功表达了VP3蛋白,对于研究VP3蛋白的免疫原性和生物学特性奠定了的基础。

Objective To construct proeukaryotic expressive vector of VP3 gene of JC virus, and observe the expression of recombinant protein. Methods The DNA fragment of vp3 was amplified by polymerase chain reaction （PCR）, and cloned into pGEM-T vector. After sequencing, the correct DNA fragment was inserted into inducible proeukaryotic expressive vector pET-32a（＋） and transformed into E. coli BL21. The protein was induced with IPTG and analyzed with sodium dodecylsulfate-polyacrylamide gel electrophoresis and Western blot hybridization. Results The DNA fragment of vp3 was amplified by PCR. The expressive vector was constructed successfully. After induction with IPTG, recombinant target protein with Mr 59 ku was expressed. Western blot analysis showed that the protein had good antigenicity. Conclusien The recombinant vp3 gene was expressed successfully. These results lay the foundation for studying the immunogenicity and bionomics of VP3 protein.