人血管生成素基因的克隆及表达

利用RI-PCR方法从培养的人黑色素瘤细胞系A375中扩增得到了人血管生成素cDNA片段,测序正确后克隆入表达载体pET-28a( )中并转化于E.coli BL21宿主菌中.经IPTG诱导,表达了N端融合6个组氨酸标签(6His-tag)的血管生成素融合蛋白.利用6His-tag与过渡态金属离子Ni2 高亲和力结合的性质,经镍柱纯化,获得了高纯度的血管生成素融合蛋白,为进一步研究其生物活性及应用奠定了基础.

Cloning and expression of human angiogenin gene

In order to obtain much more purified human angiogenin,we expressed it in E.coli BL21.The angiogenin was amplified by PCR method from the human melanoma cell line A375 and inserted into the expression vector pET-28a( ) after it was proved to be correct by DNA sequencing.Then the constructed gene was transformed into E.coli BL21 host bacteria.After having been induced by IPTG,the recombinant protein fused with 6His-tag at its N-terminal was expressed successfully.Owing to possessing 6His-tag,the expressed protein can be purified by Ni2 resin according to the high affinity of 6His-tag with Ni2 .