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目的 :构建含乙肝病毒表面抗原前S1,S2 (HBVpreS1,preS2 )优势B细胞表位与乙肝病毒核心抗原 (HBcAg)的嵌合蛋白 ,探索其作为兼具预防和治疗HBV感染作用的新型疫苗的可能性。方法 :利用分子克隆技术先后将HBVpreS1(2 1～ 4 7AA .) ,preS2 (133～ 14 5AA .)表位基因插入HBcAg基因中 ,得到HBVC14 4 ,CS1,CS1S2融合基因 ,分别克隆到原核表达载体pQE_30中 ,在大肠杆菌 (E .coli)M15中进行表达 ,用Ni2 + 固相化的螯合SepharoseFastFlow亲和层析纯化重组蛋白 ,最后进行抗原性的鉴定。结果 :构建了HBV嵌合型颗粒蛋白表达载体 ,并在E .coli中高效表达出可溶性病毒样颗粒蛋白C14 4 ,CS1,CS1S2 ,经Ni_NTA亲和层析纯化后 ,蛋白纯度达 80 % ,Western印迹及ELISA证明蛋白各表位都具有抗原性。结论 :本研究为进一步深入研究新型HBV治疗性疫苗的功能和应用奠定了基础
OBJECTIVE: To construct a chimeric protein containing hepatitis B virus surface antigen pre-S1, S2 (HBV preS1, preS2) and hepatitis B virus core antigen (HBcAg), and explore its potential as a novel vaccine for the prevention and treatment of HBV infection possibility. Methods: HBVpreS1 (2 1 ~ 4 7AA.) And preS2 (133 ~ 14 5AA.) Epitope genes were successively inserted into HBcAg gene by molecular cloning technology, and the fusion genes of HBVC14 4, CS1 and CS1S2 were obtained and cloned into the prokaryotic expression vector pQE_30, expressed in E. coli M15, and purified with Ni2 + immobilized SepharoseFastFlow affinity chromatography, finally, the antigenicity was identified. Results: The HBV chimeric granule protein expression vector was constructed and the soluble virus-like particle proteins C14 4, CS1 and CS1S2 were highly expressed in E.coli. After purified by Ni_NTA affinity chromatography, the protein purity was 80% Blotting and ELISA demonstrated that each epitope of the protein is antigenic. Conclusion: This study laid the foundation for further study on the function and application of novel HBV therapeutic vaccine