猪TOLL样受体2基因在CHO-K1细胞中的表达.pdfVIP

2008年2月 甘 肃 农 业 大 学 学 报 第4 3卷 第 1期14～17 JOURNAL OF GANSU AGRICULTURAL UNIVERSITY 双 月 刊 猪TOLL样受体2基因在C H O—K 1细胞中的表达 房永祥 ，景志忠。，陈国华 ，段风云。，蒙学莲。，窦永喜。 (1．甘肃农业大学，甘肃 兰州 730070；2．中国农业科学院兰州兽医研究所，甘肃省动物寄生虫病重点实验室． 家畜疫病病原生物学国家重点实验室，甘肃 兰州 730046) 摘要：将含猪TLR2(猪Toll样受体2。pTLR2)基因的真核表达载体PcDNA3．1／CT—GFP—pTLR2，采用脂 质体法转染到CH()一K1细胞，用G418持续筛选获得了阳性细胞克隆．阳性细胞克隆系经 PCR和RT—PCR检测 表明：pTLR2基因得到稳定整合并能成功地转录．荧光显微镜观察．转染细胞可见绿色荧光，表明获得了表达 pTLR2的克隆细胞系． 关键词：猪Toll样受体 2；CH()一K1细胞；转染 中图分类号：Q 786 文献标识码：A 文章编号：1003—4315(2008)01-0014—04 Expression of porcine Toll—like receptor TLR2 gene in the CHO—K1 cells FANG Yong—xiang ，JING Zhi—zhong。，CHEN Guo—hua。，DUAN Feng—yun。， MENG Xue—lian。。DOU Yong—xi。 (1．Gansu Agricultural University．Lanzhou 730070．China；2．Key Laboratory of Veterinary Parasitology of Gansu Province．State Key Laboratory of Veterinary Etiological Biology．Lanzhou Veterinary Research Institute．CAAS．Lanzhou 730046．China) Abstract：The recombinant plasmid PcDNA3．1／CT—GFP—pTLR2 was transfected into the CH0一 K1 cells by lipofectin and the positive cell clone was obtained after screening by G418 for 2～3 weeks．A— nalysis of the exogenous gene by PCR and RT‘——PCR indicated that the pTLR2 gene was integrated into the CHO—K1 cells．The expression of pTLR2一GFP fusion protein was observed by intensity fluorescence microscope at different time after transfection．The results indicated that pTLR2一GFP fusion protein was successfully expressed in the ceils and these ceils could be further used to investigate the functions of pTLR2． Key words：porcine Toll—like receptor 2；CHO—K1 cells；transfection Toll蛋白最初是在研究果蝇胚胎发育中被发 过程中起重要作用．Toll样受体(Toll—like recep— 现的，随后证实是一个广泛存在于昆虫、脊椎