乙型肝炎病毒核心蛋白c／e1表位处TC标签的基因标记对S和e抗原表达的影响

Insertion of TC motif into hepatitis B virus core protein c/e1 site does not affect the expression of S and e antigen

目的研究乙型肝炎病毒核心蛋白c／e1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HBVC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／e1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，Western blot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／e1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。

Objective To investigate whether insertion of TC motif into hepatitis B virus （HBV） core protein c/e1 site affects the expression of S and e antigen. Methods Different oligonucleotides encoding TC motif were inserted into the c/e1 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 of TC motif into HBV core protein c/e1 site does not interference with the expression of viral protein encoded by HBV genome.