FAS配体(FASL)在COS-7细胞中的表达  

Expression of recombinant FAS ligand in COS 7 cells

作　　者：罗振革[1] 彭虹[1] 孔祥英[1] 高杰英[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100850

出　　处：《中国免疫学杂志》1999年第9期385-387,共3页Chinese Journal of Immunology

摘　　要：目的：获得ＦＡＳＬ的真核细胞表达，并分析其功能。方法：将编码人ＦＡＳＬ（ＦＡＳｌｉｇａｎｄ） 的全长ｃＤＮＡ 插入真核细胞瞬时表达载体ＰＳＶＬ的ＳＶ４０ 晚期启动子下游的ＸｂａＩ位点，构建ＦＡＳＬ 的真核细胞表达载体ＰＳＶＬｈＦＡＳＬ，用电击法将ＰＳＶＬｈＦＡＳＬ转染ＣＯＳ７ 细胞，转染后４８ ｈ ，用Ｎｏｒｔｈｅｒｎｂｌｏｔ 法检测ＦＡＳＬ的表达。结果：转染后ＣＯＳ７ 细胞有ＦＡＳＬ的ｍＲＮＡ 的表达，在ＣＯＳ７ 细胞上表达的重组ＦＡＳＬ能诱导ＦＡＳ阳性的Ｕ９３７ 细胞发生程序性死亡。结论：在ＣＯＳ７ 细胞表面表达的重组ＦＡＳＬ分子能用于进一步功能研究和ＦＡＳ系统拮抗剂的筛选。Objective:Obtain recombinant FASL molecules in COS 7 cell line and study the functions of FASL Methods:The cDNA for human FAS ligand(FASL) was cloned into the downstream of SV40 late promoter of PSVL plasmid, which is a eukaryotic transient expressive vector By electroporation, PSVL hFASL was transfected into cos 7 cells.48 h after transfection,whole RNA of transfected cos 7 cells was prepared and analyzed by Northern blot.Results:PSVL hFASL transfected cells had the expression of human FAS ligand mRNA, PSVL mock transfected cells was negative There is expression of FAS antigen on the surface of U937 cells With U937 cell line as target, the activity of recombinant human FAS ligand (rhFASL) was assayed After coculture of U937 cells with FASL transfected COS 7, the target cells appeared apoptosis phenomenom Conclusion:Recombinant FasL on the surface of COS 7 cells can be used as functional study

关 键 词：FASL 真核表达 程序性细胞死亡 COS-7细胞 

分 类 号：R392.12[医药卫生—免疫学]