分泌蛋白编码基因THEM6克隆表达及组织细胞分布  

THEM6 Expression and Distribution in Different Cell Lines and Tis- sues

作　　者：李红敏[1] 任慧[1] 乔雍[1] 郝晓花[1] 刘燃[1] 王智强[1] 魏红山[1] 

机构地区：[1]首都医科大学附属北京地坛医院传染病研究所,北京市100015

出　　处：《医学分子生物学杂志》2013年第4期193-197,共5页Journal of Medical Molecular Biology

基　　金：资助项目：国家自然科学基金（No.81071411,81271901）

摘　　要：目的体外克隆表达THEM6，诱导重组蛋白，制备多克隆抗体，明确在不同组织细胞的表达特征。方法以外周血单核细胞总RNA反转录得到的cDNA为模板，扩增THEM6基因片段，诱导剂IFrG诱导THEM6重组蛋白表达，制备多克隆抗体，分析特异性、检测效价。用CCK-8检测试剂盒，分析THEM6重组蛋白对Jurkat细胞增殖的影响；观察该蛋白的组织、细胞系表达分布。结果成功体外克隆THEM6．获得重组蛋白及抗THEM6的多克隆抗体，ELISA （enzyme-linked immuno sorbent assay）酶联免疫吸附测定检测效价〉1280000；低浓度THEM6重组蛋白对Jurkat细胞增殖有影响，THEM6蛋白在Jurkat细胞等多种细胞及淋巴、肝脏等多种组织中表达。，结论THEM6基因在多种组织细胞中有广泛表达。Objective To express THEM6 recombinant protein in vitro, prepare the polyclonal antibody specific for THEM6 protein and examine the expression characteristics of THEM6 in differ- ent cell lines and tissues. Methods The total RNA of peripheral blood mononuclear cells was re- verse-transcripted to cDNA. THEM6 was amplified by using the cDNA as template. The protein ex- pression was induced with IPTG, and used to prepare the polyclonal antibody. The specificity and potency of the antibody were evaluated. The CCK-8 assay kit was used to detect the effect of THEM6 recombinant protein on proliferation of Jurkat cells. Western blotting was used to observe the expres- sion of the protein in different tissues and cell lines. Results THEM6 was successfully cloned in vitro. The recombinant protein of THEM6 and its polyclonal antibody were obtained. ELISA showed that the titer of the polyclonal antibody was 〉 1 ： 1280000. The THEM6 recombinant protein at low concentrations （1-20 ng/mL） could promote the proliferation of Jurkat cells. The THEM6 protein was found to express in Jurkat cells and many other different cell lines and tissues. Conclusions THEM6 is extensively expressed in many different cell lines and tissues.

关 键 词：THEM6基因 乙型肝炎病毒 重组蛋白 多克隆抗体 

分 类 号：R373[医药卫生—病原生物学]