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目的建立乙型肝炎病毒(HCV)preS1基因与截短C基因真核表达载体,并在CHO细胞中瞬时表达。方法PCR方法扩增HBV核心区羧基端部分缺失的基因片段和preS1全基因,克隆入真核表达载体pcDNA3.1(-)后测序鉴定,并通过脂质体瞬时转染CHO细胞。结果构建了真核表达载体pcDNA3.1(-)-Ct-preS1,成功转染CHO细胞,间接免疫荧光和RT-PCR证实pcDNA3.1(-)-Ct-preS1在CHO中表达截短的核心基因和preS1基因融合蛋白。结论成功构建preS1基因与截短C基因真核表达载体,可在CHO细胞中瞬时表达,为深入研究HBV基因免疫奠定了基础。
Objective To establish an eukaryotic expression vector for hepatitis B virus (HCV) preS1 gene and truncated C gene and express it transiently in CHO cells. Methods The gene fragment deleted from the carboxyl terminus of HBV core and the preS1 gene were amplified by PCR. The recombinant plasmid was cloned into the eukaryotic expression vector pcDNA3.1 (-) and sequenced. The recombinant plasmid was transiently transfected into CHO cells by liposome. Results The eukaryotic expression vector pcDNA3.1 (-) - Ct-preS1 was successfully constructed and transfected into CHO cells. Indirect immunofluorescence and RT-PCR confirmed that pcDNA3.1 (-) - Ct-preS1 expressed truncated core in CHO Gene and preS1 gene fusion protein. Conclusion The eukaryotic expression vector of preS1 gene and truncated C gene was successfully constructed and transiently expressed in CHO cells, which lays the foundation for further study of HBV gene immunity.