乙型肝炎病毒前S1基因酵母表达载体的构建及表达

为探讨乙型肝炎病毒（HBV）前S1蛋白的功能，在真核生物酵母细胞中表达HBV前S1基因。以HBV ayw亚型全长质粒pCP10为模板，多聚酶链反应（PCR）扩增HBV前S1基因，克隆到pGEM-Tfawsk ,双酶切后回收与酵母表达质粒pGBKT7连接，将重组载体转化酶母细胞AH109，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，结果成功地构建了HBV前S1基因酵母表达载体，Western免疫印迹显示HBV前S1在酵母细胞中表达，表达产物在胞内存在，分子量30kD左右。表明HBV前S1蛋白在酵母细胞中表达成功。

CLONING AND EXPRESSION OF PRES1 GENE OF HEPATITIS B VIRUS IN YEAST

To investigate the potential role of hepatitis B virus(HBV) preS1 protein in mediating HBV adhesion to liver cell, we prepared recombinant proteins of HBV preS1 in yeast. PCR was performed to amplify the gene of HBV preS1 from the plasmid pCP10/HBV ayw subtype containing the whole fragment of HBV and the PCR product was cloned into pGEM T vector. The gene of HBV preS1 was cut from pGEM T vector and cloned into yeast expression plasmid pGBKT7, the pGBKT7 plamids containing preSl were transformed into yeast cell AH109. The yeast protein was isolated and analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) and Western blotting. The results showed that the presence of HBV presl proteins in yeast cells was confirmed by Western slot analysis. the molecular weight of the expressed product was about 30000 Da. The findings indicated that HBV preS1 was successfully expressed in yeast system.