肝细胞靶向性系统表达载体的构建  

Construction of the expression vector of PHSA-R with Env protein

作　　者：王传玺[1,2] 韩金祥[1,2] 鲁艳芹[1,2] 高雪芹[1,2] 张翠[1,2] 

机构地区：[1]山东省医药生物技术研究中心 [2]卫生部生物技术药物重点实验室,济南250062

出　　处：《生物技术通讯》2004年第5期429-432,共4页Letters in Biotechnology

基　　金：国家自然科学基金(30371328);山东省自然科学基金(Q99015)

摘　　要：在肝脏疾病的基因治疗过程中,为把目的基因定向导入靶细胞,构建了具有靶向性的逆转录病毒的包装细胞,即应用RT-PCR方法分别反转录并扩增env、pres2基因,将它们分别克隆至pGEM-T载体上,经测序正确后,将一目的基因亚克隆在pcDNA3.1(-)表达载体上,然后将另一目的片段从T载体上切下,克隆至经同样酶切的重组表达载体中。从而成功地构建了肝细胞靶向性系统的表达载体pcDNA3.1(-)-env-pres2和pcDNA3.1(-)-pres2-env。The env?pres2 gene coding Env protein and polymerized human serum albumin receptor(PHSA-R) were amplified by RT-PCR, they were cloned respectively into vector pGEM-T. After DNA sequences were determined, one insert gene was subcloned into expression vector pcDNA3.(-), then the other insert gene which was digested by two restriction endonucleases was subcloned into recombined plasmid with the same sites, the fusion expression vectors were named pcDNA3.1(-)-env-pres2 and pcDNA3.1(-)-pres2-env.

关 键 词：靶向性 肝细胞 PCDNA3 表达载体 pres2基因 包装细胞 逆转录病毒 T载体 目的基因 克隆 

分 类 号：Q78[生物学—分子生物学] R730[医药卫生—肿瘤]