PNP表达质粒的构建及鉴定

The Construction and Identification of a Vector Containing PNP Suicide Gene

目的构建并鉴定一种新型的PNP自杀基因表达载体。方法将PNP基因插入到pcDNA3.0中,构建PNP基因表达载体pcDNA3.0/PNP;通过酶切、PCR及测序鉴定重组体。采用脂质体介导法将pcDNA3.0/PNP转染三种肿瘤细胞株,RTPCR检测PNP基因在各细胞株中的表达。结果目的片段均成功插入相应的载体中,CMV启动子调控下的PNP基因在不同的肿瘤细胞株中实现了表达。结论CMV启动子调控的PNP基因表达载体是肿瘤基因治疗中一种新型、高效的治疗载体。

Objective To construct and identify a novel expression vector harboring PNP gene. Methods Inserting PNP gene into pcDNA 3.0, an expression vector containing PNP gene driven by the CMV promoter, pcDNA 3.0/PNP was constructed by using recombinant DNA techniques. The recombinant was analyzed and identified by restriction enzyme, PCR and sequencing. The expression vector carrying PNP gene was transfected into three tumor cell lines by liposome-mediated method. The expressions of PNP gene in all cell lines were detected by RT-PCR method. Results The target fragment was cloned into the corresponding vector. The expressions of PNP gene in different tumor cell lines were positive. Conclusion The expression vector containing PNP gene under the control of a CMV promoter, is an novel effective expression vector for tumor gene therapy.