丙型肝炎病毒E1基因在COS-7中的表达

Expression of HCV E1 Gene in COS-7 Cells

目的 :了解丙型肝炎病毒 (HCV)E1基因在真核细胞中的表达情况。方法 :采用基因重组技术 ,构建含HCVE1基因的重组表达质粒pEF1/HisC E1,经酶切和PCR鉴定后 ,用脂质体转染法将重组质粒导入COS 7进行表达 ,经蛋白印迹实验鉴定表达产物。结果 :重组表达载体pEF1/HisC E1在COS 7中获得表达。表达的E1糖蛋白约 2 9kD ,具有抗原性。结论 :HCVE1基因在真核细胞高效表达 。

Objective:To understand the characteristic of expression of HCV E1 gene in eukaryotic cell.Methods:The recombinant expressive vecter pEF1/His C E1 which harbouring HCV E1 gene was constructed.After identifying by enzyme digestion and PCR,the recombinant plasmid was transfected into COS 7 cells using Lipofectamino regents.The cell lysates were analyzed by Western blots.Results:HCV E1 gene was expressed in COS 7 cells,which yields 29 kD glycoprotein,and this glycoprotein can react with HCV(+) sera.Conclusion:HCV E1 gene can be highly expressed in eukaryotic cell,which lay the basis to further study HCV E1 glycoprotein.