摘要
目的 构建HBVX HCVC融合基因真核表达载体 ,并获得稳定表达该基因的HepG2细胞株。方法 双酶切质粒pXT1 X ,得到完整的HBVX基因片段后 ,将其插入到质粒PBK CMV和PBK HCVC的相应酶切位点 ,得到重组质粒PBK X和PBK X C ;再将质粒RBK CMV、PBK X、PBK HCVC和PBK X C分别导入肝癌细胞株HepG2中 ,G418筛选 ,RT PCR、蛋白印迹鉴定HBVX和HCVC蛋白表达。结果 质粒PBK CMV、PBK X、PBK HCVC和PBK X C在HepG2细胞中有稳定表达。结论 成功构建HBVX HCVC融合基因真核表达载体 。
ObjectiveTo construction of HBV X-HCV C gene coexpression vector and get the expressed HepG2 cell line.MethodsThe HBV X gene was recovered by enzyme excision,and cloned into PBK-CMV and PBK-HCV C, get the recombined plasmids PBK-X and PBK-X-C The plasm id s PBK-CMV, PBK-X, PBK-HCV C and PBK-X-C were transfected into HepG 2 cells with liposome.After selected with G418, resistant colonies were obtained.T he reverse transcription PCR and western blot was analyzed to show HBV X an d HCV core protein expressed.ResultsThe recombinant plasmid PBK-X- C could express HBV X and HCV core protein efficiently under the control o f ve ctor's promoter.ConclusionWe have constructed HBV X-HCV C gene coexpression vector and get the expressed HepG2 cell line.
出处
《胃肠病学和肝病学杂志》
CAS
2003年第5期419-421,共3页
Chinese Journal of Gastroenterology and Hepatology
基金
国家自然科学基金资助项目 (No .3990 0 1 76 )
