西安交通大学学报（医学版）2013，Vol.34Issue(4)：454-459,6.
miRNA-7真核表达载体的构建与鉴定
Construction and identification of eukaryotic expression vector encoding miRNA-7
摘要
Abstract
Objective To construct a eukaryotic expression vector encoding miR-7 and detect its effect on the growth of human lung cancer cells. Methods Pri-miR-7 sequence was amplified from genomic DNA of human 95D cells by PCR and subcloned into eukaryotic expression vector pcDNA3. l(-) using restriction site of BamH Ⅰ and HindⅢ . The positive recombinant pcDNA3. 1 (-)-pri-miR-7 (p-miR-7) was identified by enzyme cleaving and sequencing. p-miR-7 was transiently transfected into 95D cells and the expression level of miRNA-7 was determined by Real-time PCR using specific probe before and after transfection. The proliferation of 95D cells was also accessed by MTT assay. Results Eukaryotic expression vector encoding miR-7 (p-miR-7) was identified as correct by enzyme cleaving and sequencing. The expression level of miR-7 in 95D cells transiently transfected with p-miR-7 was increased significantly (P<0.05). The proliferation of 95D cells in vitro was markedly inhibited (P<0.05). Conclusion The eukaryotic expression vector pcDNA3.1(-)-pri-miR-7 is successfully constructed, which provides the experimental foundation for further studies on the role and mechanism of miR-7 in the pathogenesis of lung cancer.关键词
miR-7/真核表达/肺癌/MTT法Key words
miR-7/ eukaryotic expression/ lung cancer/ MTT assay分类
医药卫生引用本文复制引用
宋永祥,李颖,秦安东,廖珍媛,李永菊,罗军敏,任涛,徐林..miRNA-7真核表达载体的构建与鉴定[J].西安交通大学学报（医学版）,2013,34(4):454-459,6.基金项目
贵州省优秀科技教育人才省长专项资金 (No.2009C457) （No.2009C457）
贵州省科技技术厅项目(No.2009C491) （No.2009C491）
遵义医学院博士启动基金项目(No.2008F329) （No.2008F329）
