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目的 构建HBV核心蛋白和B7.1分子真核表达嵌合质粒 ,并检测其在体外的表达。方法 利用亚克隆技术 ,将HBV核心基因片段和B7.1基因片段构建至真核表达质粒 pcDNA3中 ,测定序列后 ,用脂质体包裹转染细胞 ,采用间接免疫荧光法检测两种蛋白的表达。结果 核酸序列测定证实实验所构建的质粒正确 ,嵌合质粒中所含的HBcAg和B7.1分子的核苷酸序列与HBVadr亚型标准株核心区及鼠B7.1分子的同源性分别为 98.18%和 99.89%。该嵌合质粒在体外转染细胞内可表达HBcAg和B7.1分子。 结论 实验所构建的HBcAg/B7.1嵌合质粒能在体外同时表达HBcAg和B7.1分子。
Objective To construct chimeric plasmids for eukaryotic expression of HBV core protein and B7.1 and to detect their expression in vitro. Methods The HBV core gene fragment and B7.1 gene fragment were subcloned into the eukaryotic expression plasmid pcDNA3 using subcloning technique. After sequencing, the transfected cells were transfected with liposomes and the expression of two proteins were detected by indirect immunofluorescence . Results Nucleic acid sequence analysis confirmed that the constructed plasmid was correct. The homology of the nucleotide sequences of HBcAg and B7.1 contained in the chimeric plasmids with the core region of HBVadr subtype standard strain and the mouse B7.1 molecule were 98.18% and 99.89%. The chimeric plasmid can express HBcAg and B7.1 molecules in transfected cells in vitro. Conclusion The constructed HBcAg / B7.1 chimeric plasmid can express both HBcAg and B7.1 molecules in vitro.