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我们首次将人凝血因子Ⅸ基因cDNA亚克隆于原核细胞表达载体pKK233-2,以期获得人重组凝血因子Ⅸ。实验结果,我们得到重组表达质粒pKK233-FIX1和pKK233-FIX2,以IPTG诱导,在大肠杆菌JM103中表达出一特定的、与人凝血因子Ⅸ基因cDNA表达相关的蛋白质。ELISA定性实验表明:这一特定的蛋白质具有人凝血因子Ⅸ抗原性。此项研究为最终获得人重组凝血因子Ⅸ打下了基础。
For the first time, we subcloned cDNA of human coagulation factor Ⅸ gene into prokaryotic expression vector pKK233-2 to obtain recombinant human coagulation factor Ⅸ. As a result, we obtained the recombinant expression plasmids pKK233-FIX1 and pKK233-FIX2, induced by IPTG, and expressed a specific protein related to cDNA expression of human coagulation factor IX gene in E. coli JM103. Qualitative ELISA experiments show that: This particular protein has human factor IX antigenicity. This study laid the foundation for the final human recombinant factor IX.