hSOD基因在大白鼠肺上皮细胞中的表达

EXPRESSION OF HUMAN Cu, Zn_SOD GENE IN RAT LUNG EPITHELIAL CELL

本文以哺乳动物细胞表达载体ｐＲｃ／ＣＭＶ为运载体，构建了受控于ＣＭＶ启动子的ｈＳＯＤ基因的表达载体ｐＲｃ／ＣＭＶ＿ＳＯＤ，用脂质体转染法转染大白鼠肺上皮细胞Ｌ２细胞后，经抗生素Ｇ４１８选择性培养，获得与细胞染色体ＤＮＡ稳定整合的ｈＳＯＤｃＤＮＡ导入细胞，经ＳｏｕｔｈｅｒｎＢｌｏｔｓ和ＷｅｓｔｅｒｎＢｌｏｔｓ分析，结果表明，ｈＳＯＤ在细胞中得到稳定表达，同时测定细胞ＳＯＤ酶活性，ｈＳＯＤｃＤＮＡ导入细胞的ＳＯＤ酶活性是空载体导入细胞和父本细胞的约２倍，并且在选择细胞克隆之后的６０天内是持续的．

Expression plasmids containing hSODcDNA have been constructed using mammalian cell expression plasmid pRc/CMV. The expression of the gene isunder control of CMV promoter.The constructed expression plasmids were transfected into L2 cells by Lipofection. The transfected cells were selected by inclusion of G418 in the culture medium.The introduction of hSODcDNA in the transfected cells was confirmed by Southern Blots and Western Blots.SOD activity in the transfected cells was enhanced for 60 d after the selection of cell clones.