中文摘要：

目的构建人HIF-1α的mi R RNAi慢病毒载体。方法利用已经构建成功的针对人HIF-1α基因的mi RNA干扰质粒pcDNATM6.2-GW/EmGFP-HIF-1αmi R，通过BP/LR反应，将EmGFP-HIF-1α-mi R表达框整合重组到产毒质粒pLenti6/V5-DEST中测序鉴定，用PLP1、PLP2和PLP/VSVG质粒共转染包装293FT细胞产毒，并利用GFP蛋白表达水平进行滴度测定。结果经测序鉴定证实成功构建针对人HIF-1α基因的mi RNA慢病毒干扰载体pLenti6/V5-GW/EmGFP-HIF-1αmi R。结论成功构建针对人HIF-1α基因的mi RNA慢病毒RNA干扰载体。

英文摘要：

Objective To construct miR RNAi lentiviral vector targeting for HIF-1α of human being. Methods Based on the miR RNAi plasmid-pcDNA^TM6.2-GW/EmGFP-HIF-1α miR,the EmGFP-HIF-1α-miR cassette was reconstructed into pLenti6/VS- DEST and the virus produced plasmid was identified by sequencing. Then to transfect the 293FT cells mediated by lipofectamin 2000 and ViraPowerTM packaging plasmids （PLP1,PLP2 and PLP/VSVG plasmids） so as to produce the lentivirus. To titer the lentivirus by means of detecting GFP expression level. Results Sequencing confirmed that the miR RNAi lentiviral vector targeting for HIF-1α of human being-pLenti6/VS-GW/EmGFP-HIF-1α miR was successfully constructed. Conclusion The miR RNAi lentiviral vector targeting for HIF-1α was successfully constructed.