文档介绍：大鼠FasL基因重组慢病毒载体三质粒包装细胞系统的建立
作者:李望,孙晓青,胡书群,裴东生,陈波
【关键词】基因表达
摘要:目的建立真核细胞表达的大鼠FasL基因重组慢病毒载体三质粒包装细胞系统,为进一步研究转FasL基因在同种异体器官移植中诱导免疫耐受、保护移植物的作用奠定基础。方法制备完整的重组慢病毒载体三质粒系统:转移质粒(pLO134-FasL),包装质粒(ΔNRF)及包膜蛋白质粒(VSV-G)。脂质体法将三质粒共转染包装细胞293T,72h后收集病毒上清,Western-blot法检测293T细胞的FasL蛋白表达。结果转染后的293T细胞表达FasL蛋白。结论成功建立重组慢病毒载体的三质粒包装细胞系统。
关键词:FasL;基因表达;慢病毒载体;293T细胞
Abstract:Objective To establish the three-plasmid packaging cell line of the binant lentiviral vector encoding rat FasL gene for eukaryotic expression to meet the requisition for deep study on the effect of FasL transgene on inducing immune tolerance and protecting allografts anic The three-plasmid binant lentiviral vector,which was made up of the vector plasmid(pLO134-FasL),the packaging plasmid(
ΔNRF)and the envelop plasmid encoding the vesicular stomatitis virus-G glycoprotein(VSV-G),was isolated and embryonic kidney293T cells were cotransfected with the three plasmids by of transfection,the viral supernatant was collected and the FasL protein expressed by293T cells was detected by Western-blot FasL protein was expressed by the cotransfected293T The three-plasmid packaging cell line system of binant lentiviral vector was essfully established.
Key words:FasL;gene expression;lentiviral vector;293T cells
基于FasL(Fas li