CD80-IgG1 Fc段融合蛋白真核表达载体的构建及表达

目的：构建人CD80-IgG1 Fc段融合蛋白的真核表达载体CD80-IgG1 Fc／pcDNA3．1（＋），并住CHO细胞中进行表达。方法：应用T—A克降技术和亚克降技术，构建人CD80膜外段-IgG1 Fc段融合蛋白的真核表达载体CD80-IgG1 Fc／pcDNA3．1（＋），并将其转染人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-IgG1 Fc／pcDNA3．1（＋），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-IgGI Fc段融合蛋白的真核表达载体CD80-IgGI Fc／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。

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

AIM: To construct an eukaryotic expression vector of CD80-IgG1 Fc, and to express the fusion protein in CHO cells. METHODS: The gene encoding the CD80-IgG1 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-IgG1 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-IgG1 Fc/pcDNA3.1( )was successfully constructed and stably expressed in CHO cells, providing basis anti-tumor study.