肿瘤转移抑制基因KISS-1和NM23-H1的原核表达与纯化

Prokaryotic Expression and Purification of Metastasis Suppressor Gene KISS-1 and NM23-H1

从人胎盘组织和乳腺癌细胞中克隆得到肿瘤转移抑制基因KISS-1和NM23-H1,经Blast比对分析与GenBank公布序列同源性达100%。NM23-H1在大肠杆菌中成功表达,大部分为可溶性蛋白,占总蛋白50%以上,经纯化后纯度达到90%;而KISS-1需要与载体的部分序列融合才能够得到有效表达,表达产物为不可溶的包涵体蛋白,占包涵体总量的70%以上,经纯化后KISS-1蛋白可能形成多聚体结构。

Metastasis suppressor gene KISS-1 and NM23-H1 were cloned from human placenta and breast cancer cell, share 100% identity with that published in GenBank. NM23-H1 was successfully expressed in E.coli, the major amount of protein was soluble, accounting for more than 50% of total protein, the purified protein has 90% purity ; however, KISS-1 should be fused to the sequence from expression vector then can effectively expressed, the expressed protein was insoluble inclusion body, accounting for more than 70% of total insoluble protein, after purification, KISS-1 may formed into polymeric structures.