FasL N-端编码序列对其表达的影响  

Fas ligand N-terminal encoding sequence affectes it's self expression

作　　者：李健[1] 郑德先[1] 刘彦信[1] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2005年第7期624-627,共4页Basic and Clinical Medicine

基　　金：国家自然科学基金(39570327)

摘　　要：目的探讨Fas配体(Fas ligand, FasL)胞内区氨基酸组成对膜型FasL蛋白表达的影响.方法 PCR技术扩增FasL胞内区产生不同缺失的cDNA,构建重组表达质粒;使用脂质体转染法将表达质粒导入靶细胞;RT-PCR分析FasL mRNA水平;荧光抗体染色、流式细胞仪和Western免疫印迹技术检测细胞中FasL蛋白的表达.结果 FasL N-端缺失第2～16氨基酸残基后,FasL蛋白在细胞中的表达水平显著增加,但mRNA水平不变.结论 FasL N-端第2～16氨基酸残基对FasL蛋白表达水平具有自我调节作用.Objectives To study the expression and regulation of FasL. Methods A series of recombinant plasmids encodingthe full length of FasL as well as the mutants with various deletions at FasL N-terminal were constructed in the eukaryotic expression plasimd. Expressions of these recombinant constructs were carried out in 293E human epithelial cells.The levels of FasL protein and mRNA were detected by flow cytometry, Western blot, and RT-PCR. Results It was shown that the mutants with the deletion of 2 - 16 a.a. residue at FasL N-terminal caused markedly augment of FasL protein level both in the ceils and on the cell surfaces, but not FasL mRNA level in the ceils. Conclusions Fas ligand N-terminal encoding sequence could auto-regulate its protein expression.

关 键 词：FASL 胞内区 表达调控 

分 类 号：R346[医药卫生—基础医学]