携带绿色荧光蛋白基因的逆转录病毒载体的构建及其介导的T细胞基因转移研究.pdfVIP

Journal of Expermi ental Hem atology 2005; 13( 4): 64 1- 644 # 641# (Article ID): 1009- 2137(2005)04- 0641- 04 # 论著# T * , , , , , 徐州医学院附属医院血液科, 徐州 221002 ( green fluorescen t protein, GFP) T , ( p osp oglycerate k inase promoter, PGK ) GFP cDNA pLXSN, PA317 , G 418 , N IH 3T3T , GFP 。; PA 317 , GFP。G 418, GFP T。: 、T, T 。 ; ; T; R392. 4; Q755; Q782 A Construction of theRetroviralVector Carrying theGreen F luores2 cent Protein Gene and ItsApplication in ediatingGeneTransfer intoT Cells CHEN XiangM2 ei, XU Kai2Lin, PAN Xiu 2Ying, LI Z en 2Yu, LU Qun 2Xian, LIDe2P eng D eparmt ent ofH ematology, t e Affilia ted H ospital ofXuz ou M edical College, Xuz ou 221002, C ina Abstract To con struct retroviral vector carrying green fluorescent protein (GFP) gene, and determ ine w et er T cells can be infected by retrovirus, t e p osp oglycerate kinase ( PGK) promoter gene and GFP full2lengt cDNA w ere subcloned into t e retroviral vector pLX SN. T e recomb inan t vector was transfected into PA317 packaging cells by DNA2 calcium p osp ate coprecipitation. T e G418 resistant clonesw ere selected, t en N IH 3T3 cells and T cellsw ere in fected by t e culture supernatan t of t e retrovirus con tain ing GFP. T e resu lts s owed t atGFP expression in packaging cell line PA317 was detectab le under fluorescencem icroscope, w ic demon strated t at t eGFP retrov irusw ere ab le to in fect T cells. It is concluded t e retrovirus vectors can in troduce a foreign gene into T cells efficiently and can be used as mi portant tools to deliver gene into T cells. Key words retroviral vector; green fluorescent protein gene;