CD80-IgG1Fc段融合蛋白真核表达载体的构建及表达  

Construction of eukaryotic expression vector of CD80-IgG1 Fc fragment fusion protein and its expression in CHO cells

作　　者：李和军[1] 李频[1] 周小玲[1] 郑祥雄[1] 

机构地区：[1]福建医科大学附属协和医院临床免疫研究所,福建福州350001

出　　处：《细胞与分子免疫学杂志》2006年第1期40-42,46,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：福建省科技厅基金资助项目(No.2003D06)

摘　　要：目的构建人CD80-IgG1Fc段融合蛋白的真核表达载体CD80-IgG1Fc/pcDNA3.1(+),并在CHO细胞中进行表达。方法应用T-A克隆技术和亚克隆技术,构建人CD80膜外段-IgG1Fc段融合蛋白的真核表达载体CD80-IgG1Fc/pcDNA3.1(+),并将其转染入CHO细胞株并筛选,进行稳定表达。通过Western blot及ELISA法,检测融合蛋白的表达。结果成功地构建了真核表达载体CD80-IgG1Fc/pcD-NA3.1(+),并建立CHO细胞稳定表达株。Western blot及ELISA法检测到细胞培养上清中有融合蛋白的表达。结论成功地构建了人CD80膜外段-IgG1Fc段融合蛋白的真核表达载体CD80-IgG1Fc/pcDNA3.1(+),并在CHO细胞中稳定表达,为下一步抗肿瘤的研究奠定了基础。AIM ： To construct an eukaryotic expression vector of CD80-1gGI Fc, and to express the fusion protein in CHO cells. METHODS： The gene encoding the CD80-1gGI Fc fusion protein were constructed in eukaryotic expression vector pcDNA3. 1 （ ＋ ） by means of T-A cloning and subcloning techniques, then was transfected into CHO cells for stable expression. The expression of the fusion protein was detected by Western blot and ELISA. RESULTS： DNA sequencing and restriction endonuclease digestion analysis indicated that the eukaryotic expression vector CD80-1gGI Fc/ pcDNA3. 1 （ ＋ ）was successfully constructed. After the recombinant plasmid was transfected into CHO cells, the stable expression of the fusion protein was demonstrated by Western blot and ELISA. CONCLUSION： The eukaryotic expression vector of CD80-1gGI Fc/pcDNA3.1 （ ＋ ） was successfully constructed and stably expressed in CHO cells, providing basis anti-tumor study.

关 键 词：CD80 IGG1 FC段 融合蛋白 基因克隆 真核表达 

分 类 号：R392.11[医药卫生—免疫学]