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.
出处
《中华实验和临床病毒学杂志》
CAS
CSCD
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
关键词
CDNA
重组逆转录病毒
转染
感染
基因治疗
Tissue-type plasminogen activator
Recombinant retrovirus
Transfection
Infection
