反转录病毒载体介导可溶性Fas的体外表达  

Retroviral Vector-Mediated In Vitro Expression of Human Soluble Fas

作　　者：王良利[1] 邹萍[1] 胡中波[1] 刘凌波[1] 李爱香[1] 马艳萍[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,武汉430022

出　　处：《中国实验血液学杂志》2002年第2期97-99,共3页Journal of Experimental Hematology

基　　金：为国家自然科学基金资助项目编号C0 30 30 81 1

摘　　要：将含全长FascDNA的质粒利用PCR技术进行特定片段删除 ,获得sFascDNA。通过DNA重组技术 ,将sFascDNA插入到反转录病毒载体 pLXIN ,构建了重组表达载体pLXIN sFas。经酶切鉴定及测序 ,表明成功地构建了重组表达载体 pLXIN sFas。该载体经PA317细胞包装后 ,感染靶细胞COS 7,分别采用ELISA及凋亡抑制实验检测其蛋白表达量为 (2 .2± 0 .7) μg/ml,且具有良好的生物学活性 ,能明显抑制抗 Fas(CH 11)介导的T淋巴瘤细胞株Jurkat的凋亡。Leukemic cells from patients expressed high level FasL cause apoptosis of autologous activated T cells via the Fas/FasL pathway. To investigate the role of soluble Fas(sFas) in reversing this process, a retroviral mediated expression vector pLXIN sFas was established. A retroviral mediated expression system of human sFas was established in vitro and the bilogical activity of the expression product sFas was observed. To obtain the soluble Fas cDNA, the specific part of the full length Fas cDNA was deleted by multiple PCR. After pLXIN sFas packaged by PA317 cells, it was transferred into the target cell COS 7. The quantity of the sFas was (2.2±0.7) μg/ml in supernatant of cultured COS 7 cells, and it could greatly inhibit apoptosis of Jurket cells induced by anti Fas antibody. Our results suggested that the recombinant is able to express the target proteins in vitro and it has the perfect biological activity.

关 键 词：反转录病毒载体 可溶性FAS 基因表达 基因转移 体外表达 白血病 免疫监视 

分 类 号：R733.7[医药卫生—肿瘤]