四种乙肝核酸疫苗质粒的构建及在真核细胞中的表达

Construction of Four Kinds of Recombinant HBV DNA Vaccine and Their Expression in Eukaryotic Cells

目的:构建四种乙肝核酸疫苗质粒pCI/S、pCI/S2S、pCI/S1S2S、pCI/S2SS2,并检测其转染真核细胞后的蛋白表达。方法:从慢性乙肝患者的混合血清中制备HBV DNA模板,分别扩增和连接,得到四个基因片段S、S2S、S1S2S、S2SS2,与pCI载体进行连接,得到四个重组质粒pCI/S、pCI/S2S、pCI/S1S2S、pCI/S2SS2,将其分别转染SP2/0细胞,检测目的抗原蛋白的表达情况。结果:四种重组质粒转染细胞后,裂解液中均可检测出目的抗原蛋白的表达。结论:所构建的四种重组质粒均能在哺乳动物细胞内表达相应的目的抗原蛋白,可用于下一步动物实验。

Objective： To construct four kinds of recombinant HBV DNA vaccine pCI/S, pCI/S2S, pCI/S1S2S and pCI/ S2SS2, and the effectiveness of recombinant vaccine. Methods： DNA templates were acquired from mixed serum in patients infected with chronic hepatitis B, based on which four gene fragments including S. S2S. S1S2S. S2SS2 was cloned. Four kinds of recombinant plasmid pCI/S, pCI/S2S, pCI/S1S2S and pCI/S2SS2 were constructed. SP2/0 cells were transfected with the four kinds of recombinant plasmid, and HBsAg expression in cell split fluids was detected by ELLSA. Results： Four kinds of recombi- nant plasmid pCI/S, pCI/S2S, pCI/S1S2S and pCI/S2SS2 were in accordance with what we anticipate, nBsAg expression in cell split fluids could be detected by ELISA. Conclusion： The specific protein expression was all detected in mammal animal cells transfected with the four kinds of recombinant plasmid, and these recombinant plasmid could be used in animal test.