丙型肝炎病毒核心蛋白基因的表达载体构建及在酵母中的表达

目的：为探讨丙型肝炎病毒（HCV）核心（core）蛋白的功能，在真核生物酵母细胞中表达HCV核心蛋白基因。方法：用多聚酶链反应（PCR）的方法在HCV全长质粒pBRTM／HCV为膜板扩增HCV核心蛋白基因，克隆到pGEM－T载体中，双酶切后回收连接到酵母表达质粒pGBKT7中表达。提取酵母蛋白质，进行十二烷基磺酸钠－聚丙烯酰胺凝胶电泳（SDS－PAGE）和Western免疫印迹分析。结果：成功构建HCV核心蛋白基因酵母表达载体，Western免疫印迹显示了HCV核心蛋白在酵母细胞中表达。表达产物在胞内存在，分子量22000ku左右。结论：HCV核心蛋白在酵母中表达成功。

Cloning and expression of the gene of hepatitis C virus core in yeast

Objective:The function of HCV core is very complicated , the HCV core protein playsan important role in disease caused by HCV In the report we investigate the gene expression of HCV core in yeast Methods:PCR wasperformed to amplify the gene of HCV core from the plasmid pBRTM/HCV containing the whole fragment of HCV and the gene was cloned into pGEM T vector The gene of HCV core was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, then pGBKT7 core was transformed into yeast AH109 The yeast protein was isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting assay Results:HCV core gene was successfully cloned into pGBKT7 The results of SDS PAGE and Western blotting assay showed (1)the molecular weight of the expressed product was about 22000Da. (2) HCV core protein was existed within yeast cells Conclusion:The findings suggested that HCV core was successfully expressed in yeast system