聚合酶链反应_____潜伏膜蛋白1;聚合酶链反应;慢病毒

LMP1与绿色荧光蛋白融合基因慢病毒的制备

http://www.100md.com 《第四军医大学学报》 2005年第17期

聚合酶链反应,,潜伏膜蛋白1;聚合酶链反应;慢病毒,0引言,1材料和方法,2结果,3讨论,【参考文献】

     Production of lentivirus of LMP1 and GFP recombinant gene

    LI XianMao, ZHAO Tong， ZHU MeiGang， ZHANG SanQuan，ZHANG JinHua

    Department of Pathology, School of Basic Medicine, Southern Medical University, Guangzhou 510515, China

    【Abstract】 AIM: To produce lentivirus of latentmembrane protein 1 (LMP1) and green fluorescent protein (GFP) recombinant gene. METHODS: The fragment including all the exons of LMP1 was amplified by RTPCR and was recombined to the downstream of CMV promoter in the lentivirus vector FUGW. The three plasmids expressing lentivirus [packaging plasmid pCMV.△8.9, envelope plasmid VSVG and target plasmid FUGWLMP1] were packaged into virus packaging cell line 293FT using lipofectamine method. The virusproducing cell supernatant was harvested and concentrated. RESULTS: It was proved that the sequence of the RTPCR product was totally consistent with the data of GenBank by DNA sequencing analysis. The LMP1 cDNA fragment was cloned in the vector FUGW in the right direction and the open reading fragment of LMP1 was maintained. The three plasmids were effectively transferred into 293FT. Much green fluorescence was observed by fluorescence microscopy. CONCLUSION: The lentivirus of LMP1 and GFP recombinant gene are successfully produced. ......

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