【摘要】 目的:研究重组质粒pLMP1-p53mt中EB病毒LMP1基因和突变型p53(p53mt)基因在HNE1和HeLa细胞中的体外表达。方法:体外培养人鼻咽癌细胞株HNE1和人宫颈癌细胞株HeLa,采用脂质体lipofectAMINETM2000介导将重组质粒pLMP1-p53mtDNA转染细胞,利用逆转录聚合酶链式反应(RT-PCR)和Sourthernblot法分析p53mt基因和LMP1基因的表达情况。结果:经重组质粒pLMP1-p53mt转染的HNE1和HeLa细胞中检测到了p53mt基因和LMP1基因的表达。结论:重组质粒pLMP1-p53mt的LMP1基因和p53mt基因能分别在EDL-2启动子和CMV启动子控制下表达。该质粒可进一步用于转基因动物的制备和基因功能研究。更多还原

【Abstract】 OBJECTIVE: To study the EB virus LMP1 gene and mutant p53(p53mt) gene expressions of recombinant plasmid pLMP1-p53mt in HNE1 and HeLa cells in vitro. METHODS:Recombinant plasmid pLMP1-p53mt was transfected into HNE1 cells and HeLa cells cultured in vitro by lipofectamine ~ TM 2000.The expressions of mutation p53 and LMP1 were detected by RT-PCR and Southern blot. RESULTS:The expressions of mutation p53 gene and LMP1 gene were observed in HNE1 cells and HeLa cells which were transfected by recombinant plasmids. CONCLUSIONS:The LMP1 gene and p53mt gene of recombinant plasmid pLMP1-p53mt can express under the control of EDL-2 promoter and CMV promoter,respectively.The recombinant plasmid pLMP1-p53mt can further be applied to preparation of transgenic animals and the study of gene functions. [更多还原