【摘要】 利用RT PCR方法从人肝细胞癌HepG2细胞株中扩增得到编码人血管抑素 (angiostatin)的cDNA。将其插入毕氏酵母表达载体 pPIC9K中 ,转化毕氏酵母 ,获得了整合多拷贝血管抑素基因的菌株。发酵液经赖氨酸亲和柱纯化所得到的重组蛋白的产量为 2 5mg/L。蛋白质氨基酸序列分析结果与国外报道一致。重组蛋白特异地抑制bFGF刺激的牛主动脉内皮细胞的增殖 ,ED50 约为 3mg/L。更多还原

【Abstract】 Human angiostatin cDNA was amplified from human hepatoma cell line HepG2 using RT PCR and was cloned into pPIC9K vector. Recombinant Pichia pastoris strain with 4 copies of angiostatin gene was obtained. Recombinant protein was purified by lysine affinity Sepharose column and the finally purified angiostatin was 25 mg/L, higher than previously reported 17 mg/L. Amino acid sequence analysis revealed the identity of our protein the same with that previously reported. Recombinant angiostatin inhibited specifically the proliferation of bovine aortic endothelial cell stimulated by bFGF, with ED 50 being about 3 mg/L.更多还原