摘要目的 研究乙型肝炎病毒核心蛋白c/el表位处TC标签的基因标记对S和e抗原表达的影响,探索TC标签标记HBV病毒体的可行性.方法 以含有1.3倍HBV基因组的载体为模板,在HBV C基因中插入编码TC标签的碱基序列,得到重组的突变型HBV载体,其编码的核心蛋白的c/el表位处嵌入了TC标签.将野生型和突变型HBV载体分别瞬时转染HepG2细胞,Westernblot检测野生型和各种TC嵌合型核心蛋白的表达,酶联免疫吸附法分析病毒S和e抗原的表达.多组间比较用单因素方差分析.结果 Western blot显示各种嵌合型核心蛋白在细胞内成功表达,并且同野生型蛋白的表达水平无显著差别,酶联免疫吸附法检测表明各组细胞S和e抗原表达水平没有明显差别.结论 HBV核心蛋白c/el表位处TC标签的基因标记不会明显影响突变HBV载体表达病毒蛋白.

abstractsObjective To investigate whether insertion of TC motif into hepatitis B virus (HBV) core protein c/el site affects the expression of S and e antigen.Methods Different oligonucleotides encoding TC motif were inserted into the c/el site of the core gene of a 1.3 copy wild-type HBV genome vector.HepG2 cells were divided into several groups of cells to transiently transfect with the wild-type and mutant HBV vectors,respectively.In each group,the expression level of core protein inside cells was detected by western blotting,and the levels of S and e antigen in culture medium were analyzed by ELISA assay.Results Western blotting showed that these TC-tagged core proteins were expressed at similar level of wild-type one.ELISA assay indicated that the level of S and e antigen in culture medium of different groups were not significantly different.Conclusion Insertion ofTC motif into HBV core protein c/el site does not interference with the expression of viral protein encoded by HBV genome.