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目的 为了获得大量重组HCVE2蛋白 ,以研究E2抗体潜在的保护作用。方法 利用PCR方法从HCV基因组序列中扩增出 931bp的E2基因片段 ,经EcoRI和Sall双酶切后连接到pET 2 8a表达载体上 ,转化大肠杆菌BL2 1(DE3)菌株 ,得到重组质粒PET -E2 ,工程菌经IPTG诱导培养 ,明显表达出HCVE2蛋白 ,表达产物经固定化金属配体亲和层析纯化 ,用ELISA方法检测生物学活性。结果 表达产物主要以包涵体形式存在 ,表达量达菌体蛋白的 18%以上 ,目的蛋白具有良好的反应原性。结论 HCVE2基因的克隆与表达为进一步开展HCVE2蛋白和疫苗研究奠定了基础
Objective In order to obtain a large number of recombinant HCVE2 protein to study the potential protective effect of E2 antibody. Methods The 931bp E2 gene fragment was amplified from the HCV genome sequence by PCR and ligated into pET 2 8a vector after digested with EcoRI and Sall. The recombinant plasmid was transformed into E. coli BL21 (DE3) E2. The engineered bacteria were induced by IPTG and the expression of HCVE2 protein was clearly expressed. The expressed product was purified by immobilized metal ligand affinity chromatography and the biological activity was detected by ELISA. Results The expressed product mainly existed in the form of inclusion body, the expression level reached more than 18% of the bacterial protein, and the target protein had good reactogenicity. Conclusion Cloning and expression of HCVE2 gene laid the foundation for the further study of HCVE2 protein and vaccine