含人组织型纤溶酶原激活剂cDNA重组逆转录病毒的制备及鉴定  

Preparation and characterization of recombinant retroviral vector containing t-PA cDNA

作　　者：崔宁[1,2] 彭鲁英[1,2] 宋后燕[1,2] 韩琴琴 陈灏珠 

机构地区：[1]上海医科大学基础医学院分子遗传学研究室 [2]上海市心血管病研究所

出　　处：《中华实验和临床病毒学杂志》1997年第1期50-52,共3页Chinese Journal of Experimental and Clinical Virology

基　　金：国家863和上海医科大学基因治疗基金

摘　　要：将人组织型纤溶酶原激活剂ｃＤＮＡ与逆转录病毒载体ＬＮＳＸ重组后转染病毒包装细胞ＰＡ３１７，形成完整的重组病毒颗粒，电镜下重组病毒呈散在分布，球形，直径９０～１８０ｎｍ，由囊膜、外壳和核心三部分组成。重组病毒颗粒感染ＮＩＨ３Ｔ３细胞，在含Ｇ４１８培养基中筛选培养２周后计数阳性细胞克隆数，结果达６×１０８ＣＦＵ／Ｌ；被感染的受体细胞ＮＩＨ３Ｔ３高效表达具有纤溶活性的重组人组织型纤溶酶原激活剂。A recombinant retroviral vector containing tissue-type plasminogen activator (t-PA) cDNA was constructed and transfected into PA317 viral packaging cells, forming intact virus particles. Under electron microscope the recombinant retroviral particles were composed of envelope, capsid and core. These viral particles were spherical with a diameter of 90～180nm, and spread dispersely in the cells. NIH3T3 cell infected by retrovirus particles were screened with G418. The virus titer of 6×10 8 CFU/L was verified by counting the positive clones two weeks after screening. The expression of t-PA was demonstrated in the NIH3T3 cells infected with the recombinant virus.

关 键 词：CDNA 重组逆转录病毒 转染 感染 基因治疗 

分 类 号：R392.33[医药卫生—免疫学] R456[医药卫生—基础医学]