AIF基因真核表达载体的构建及其在MGC-803中的表达  

Creation of AIF Eukaryotic Expression Vector and Its Expression in MGC-803

作　　者：王昌正[1] 吴本俨[1] 黄海力[1] 李英男[1] 王卫华[1] 

机构地区：[1]解放军总医院南楼,北京100853

出　　处：《中国肿瘤》2009年第7期564-567,共4页China Cancer

基　　金：解放军总医院苗圃基金(07MP35)

摘　　要：[目的]检测胃癌标本中AIF基因是否存在碱基突变;构建AIF基因真核表达载体AIF-GFP;并检测其在胃腺癌细胞株MGC-803中的表达。[方法]应用RT-PCR的方法分别从5例胃癌组织标本中提取总RNA,进一步扩增出全长的AIF基因;将其克隆到pMD19-T载体进行测序,并亚克隆至真核表达载体pEGFP-N2,重组载体AIF-GFP采用脂质体法瞬时转染MGC-803细胞,荧光显微镜观察AIF-GFP在细胞中的表达。[结果]由5例胃癌组织标本提取的mRNA所扩增出的AIF基因的序列与Gene Bank中AIF(AF100928)序列完全一致,胃癌组织中均未发现碱基突变;构建了真核表达载体AIF-GFP;并将其转染MGC-803进行表达。[结论]胃癌中AIF基因未发生碱基突变;AIF-GFP真核表达载体,在MGC-803细胞中获得了表达,为进一步探讨AIF在胃癌诊治中的应用奠定了基础。[Purpose] To analyze the gene mutation of AIF in gastric cancer samples and to creat the AIF eukaryotic expression vector AIF-GFP and to detect its expression gastric adenocarcinoma cell line MGC-803. [Methods] Total RNA was extracted from tumor tissues of 5 cases with gastric cancer, and then the full-length AIF cDNA was obtained by RT-PCR. The products were cloned into pMD19-T vector, sequenced and then subcloned into eukaryotic expressing vector pEGFP-N2. The recombined vector AIF-GFP was transfected into MGC-803 ceils. The expression of AIF-GFP was observed by fluorescent microscope. [Results ] The cDNA of AIF was sequenced and consistent with the sequence reported in Genebank （AF100928）. There is no mutation of AIF in the 5 cases. AIF eukaryotic expression vector AIF-GFP was successfully created and its expression in MGC-803 could be observed. [Conclusions] There is no gene mutation of AIF in gastric cancer, and AIF eukaryotic expression vector AIF-GFP was expressed in MGC-803 cells, which provide basis for further study of AIF on diagnosis and treatment of gastric cancer.

关 键 词：胃肿瘤 凋亡诱导因子 基因突变 基因表达 载体 

分 类 号：Q78[生物学—分子生物学] R735.2[医药卫生—肿瘤]