摘要
目的 构建人源抗 HAVFab真核表达载体并进行表达。方法采用DNA重组技术将抗体重轻链基因与信号肽基因连接 ,并分别插入真核表达载体 pCdhfr1和pCDNA3 1;以脂质体介导法转染CHO细胞 ,经G418筛选细胞 ,ELISA检测Fab基因的表达。结果成功地构建了Fab表达载体。转染细胞后 ,经ELISA检测 ,细胞培养上清中有抗原结合活性的Fab片段。结论Fab真核表达载体的构建及其在CHO细胞中的表达 。
Aim To construct a eukaryotic expression vector of human antiHAV Fab and to detect its expression in CHO cells. Methods Fd and light chain genes were ligated to their signal peptide sequences and inserted into pCdhfr1 or pCDNA3.1, respectively. The recombinant plasmids were cotransfected into CHO cells, and resistant clones were obtained by G418 selection. The expression of Fab gene was detected by ELISA. Results Recombinant plasmids were successfully constructed. The plasmidtransfected CHO cells were obtained by cotransfection and G418 selection. ELISA confirmed the expression of antiHAV Fab in culture medium. Conclusion The construction of human antiHAV Fab eukaryotic expression vector and its expression in CHO cells lay the foundation for stable expression of antibodycoding gene in CHO cells.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
2000年第3期193-195,共3页
Chinese Journal of Cellular and Molecular Immunology
基金
国家"863"课题资助!NO.Z1801
