乙型肝炎病毒X蛋白在不同肝细胞定位及表达的特性

An analysis of the features of HBx protein distributed in liver cells and its expression in E. coli.

目的探讨乙型肝炎病毒X蛋白(HBx)在肝细胞的分布特点及蛋白表达的规律。方法常规分子克隆技术构建HBx及其各种突变体的表达载体,Western blot验证HBx蛋白的表达;荧光共聚集扫描技术观察HBx蛋白在肝细胞中的分布特点;谷光甘肽S转移酶(GST)-Sepharose 4B亲和层析方法纯化GST-HBx融合蛋白。结果成功构建了全长HBx及其各种缺失突变体表达载体;HBx蛋白在肝细胞核内均匀分布,在细胞质内呈颗粒状聚集分布; 在优化蛋白表达及纯化条件下,突变体HBx(73～120 aa)的GST融合蛋白极易降解。结论为从蛋白水平探讨HBx 的生物学功能提供了有利的材料。

Objective To investigate the features of HBx protein distributed in liver cells and its expression in E.coli. Methods The expression vectors encoding the full length HBx and its mutants were constructed by the routine molecular cloning method. HBx protein expression was detected using Western blotting. The distribution feature of HBx protein in liver cells was examined using the fluorescence confocal microscopy. A series of purified HBx fusion proteins were obtained by glutathione-sepharose 4B affinity chromatography. Results The expression vectors were successfully constructed for the full length HBx and its mutants. HBx was found distributed uniformly in the nuclei but granularly in the cytoplasm of the liver cells. Under optimal conditions, the mutant GST-HBx （72-120aa） was easily degraded. Conclusion This study may provide a basis for further study on the biological function of HBx at the protein level.