重组survivin腺病毒的构建与鉴定

Construction and confirmation of a recombinant adenovirus vector of survivin

目的 构建人基因的重组survivin腺病毒载体,为喉癌的基因治疗研究提供实验基础.方法 聚合酶链反应（PCR）克隆survivin基因,PCR产物与克隆载体连接、转化,鉴定阳性克隆测序后再与腺病毒骨架质粒同源重组,对重组克隆行PCR鉴定和Xho Ⅰ、EcoR Ⅰ双酶切鉴定,并包装、纯化.结果 核酸序列分析证明,所克隆的survivin基因与文献报道一致,克隆成功,而且外源survivin基因正确地插入到腺病毒载体中,表明已成功构建腺病毒载体pLP-Ad-SUR.结论 重组survivin腺病毒载体构建成功.

Objective To construct a recombinant adenovirus of survivin vector and provid valuable reference for gene therapy of laryngeal cancer.Methods The survivin gene was cloned by PCR.After confirmation by enzyme restriction analysis and sequencing, the gene and the adenovirus vector were recombined together to construct the recombinant adenovirus vector.The recombinant adenovirus vector was confirmed via both sequencing and digestion restriction analysis, and then linearized and transfected into the HEK 293 cell line to generate recombinant adenovirus.Results The Sequence analysis demonstrated that the survivin gene sequence was the same as published in the literature, suggesting that a recombinant adenovirus vector has been successfully constructed.Conclusions A survivin recombinant adenovirus has been successfully constructed.