Lipofectin介导转移的外源性ADA基因在小鼠T淋巴细胞中的表达

Expression of Human ADA Gene in Murine T-cells after Lipofectin-mediated Gene Transfer

本文选择小鼠T淋巴细胞系（YAC-I）为基因转移的靶细胞，采用Lipofectin介导的基因转移方法，将载有人腺苷脱氨酶（ADA）基因的真核表达载体和载有Neo^R基因的真核表达载体共转化导入体外培养的YAC-I细胞，然后对转化筛选后的YAC-I细胞进行了ADA的定性定量分析。实验结果表明：经转化筛选的YAC-I细胞中的ADA活性为未转化YAC-I细胞中内源ADA活性的l．6倍，且转化筛选的YAC-I可经电泳分离出两条迁移率不同的人、鼠ADA同工酶带，提示Lipofectin介导转移的外源性人ADA基因在小鼠淋巴细胞中获得了表达，从而为以后开展基因治疗研究提供了一定的实验依据。

In this report,Murine T-Lymphocytes have been chosen as target cells for lipofectin mediated gene transfer.Eukaryotic expression vectors which carry human ADA gene and Neo^R gene respectively have been transfected into the cultured Yac-I cells by cotransfection .After selection with G418,quantitative and qualitative assays of transfected Yac-I cells were conducted.The results showed that ADA activity in transfected cells is 1.6：times higher han non-transfected cells,and two isozyme bands of human and murine ADA with different migration rates were separated from transfected and G418-selected YacI cells on acetate cellulose electrophoresis.Our results suggest that exogenous human ADA gene were expressed efficiently in murine cells.It could provide a experimental basis for further gene therapy research.