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

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.