经HIF-1α与EGFP双基因重组腺病毒转染大鼠心肌干细胞后HIF-1α基因的表达  

Expression of HIF-1α Gene in Rat Cardiac Stem Cells Transfected with HIF-1α and EGFP Double Gene Recombinant Adenovirus

作　　者：李沙[1] 李树仁[1] 郝清卿[1] 张倩辉[1] 杨玲玲[1] 

机构地区：[1]河北医科大学,附属河北省人民医院心内一科,石家庄050051

出　　处：《基因组学与应用生物学》2016年第12期3210-3218,共9页Genomics and Applied Biology

基　　金：河北省自然科学基金委员会(C2015307019)资助

摘　　要：常规经基因修饰腺病毒转染细胞后,即进行细胞的移植,并不对病毒转染效果进行检验。本研究采用HIF-1α与EGFP双基因重组腺病毒转染大鼠心肌干细胞,察看基因重组腺病毒是否能够转染心肌干细胞内;研究经HIF-1α与EGFP(enhanced green fluorescence protein,EGFP)双基因重组腺病毒转染心肌干细胞后HIF-1α基因的表达情况。在荧光显微镜下,去察看病毒转染细胞的情况,确定病毒最适MOI值;采用实时定量PCR(real-time quantitative PCR,RT-qPCR)评价HIF-1α基因表达变化;应用Western blotting方法检测HIF-1α蛋白表达变化,进而获得良好的实验结果。After transfection, the cells were transfected with gene modified adenovirus, but the effect of virus transfection was not tested. In this study, we used HIF-1α and EGFP recombinant adenovirus transfection of rat myocardial stem cells to examine whether gene recombinant adenovirus can transfect cardiac stem cells or not; To study the expression of HIF-1α gene after transfection of cardiac stem cells with HIF-1α and EGFP （enhanced green fluorescence protein, EGFP） double gene recombinant adenovirus vector. Under the fluorescence microscope, we observed virus transfected cells, viruses identified the optimal MOI value; Used quantitative real-time PCR （real-time quantitative PCR, RT-qPCR） evaluation of HIF-1α gene expression changes; By Western blotting detection of HIF-1α protein expression to gain good results.

关 键 词：心肌干细胞 基因重组 细胞转染 实时定量PCR Western blotting方法 

分 类 号：Q78[生物学—分子生物学]