重组质粒pLMP1-p53mt在人鼻咽癌HNE1和人宫颈癌HeLa细胞中的表达分析  

Analysis of recombinant plasmid pLMP1-p53mt expression in HNE1 and HeLa cells in vitro

作　　者：何迎春[1] 田道法[1] 贺安意[1] 江洁琼[1] 刘宇勤[1] 

机构地区：[1]湖南中医学院基础部生物化学教研室,湖南长沙410007

出　　处：《中华肿瘤防治杂志》2006年第6期403-406,共4页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(30271678);中国博士后科学基金第35批资助项目(2004035644);湖南省教育厅资助项目(03C287);湖南省杰出青年基金项目资助(03JJY3040)

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。OBJECTIVE： To study the EB virus LMP1 gene and mutant pS3（pS3mt） gene expressions of recombinant plasmid pLMPl-p53mt in HNE1 and HeLa cells in vitro. METHODS： Recombinant plasmid pLMP1-pS3mt was trans- fected 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 ceils and HeLa ceils which were transfected by recombinant plasmids. CONCLUSIONS： The LMP1 gene and pS3mt gene of recombinant plasmid pLMPl-pS3mt can express under the control of EDL-2 promoter and CMV promoter, respectively. The recombinant plasmid pLMPl-pS3mt can further be applied to preparation of transgenic animals and the study of gene functions.

关 键 词：HNE1细胞/代谢 HeLa细胞/代谢 质粒 细胞 培养的 

分 类 号：R739.63[医药卫生—肿瘤] R737.33[医药卫生—临床医学]