IGFBP-3高效可溶表达、纯化及生物学活性初步研究  

Soluble High-expression,Purification and Bioassay of IGFBP-3

作　　者：吴琛[1,2] 姚广印[3] 邹民吉[1] 陈光宇[1] 王旻[1] 王嘉玺[1] 徐东刚[1] 

机构地区：[1]军事医学科学院基础医学研究所 [2]河北大学生命科学学院,保定071002 [3]河北大学生命科学学院

出　　处：《生物工程学报》2007年第3期398-402,共5页Chinese Journal of Biotechnology

基　　金：国家自然科学基金资助(No.30270634)。~~

摘　　要：克隆胰岛素样生长因子结合蛋白3(IGFBP-3)的cDNA片段,构建原核表达载体pET-DsBA-IGFBP3。将该重组质粒转化大肠杆菌BL21(DE3)plysS中,诱导表达IGFBP-3融合蛋白(简称D-IGFBP3)。经检测融合蛋白主要以可溶形式表达。表达产物用His亲和层析柱纯化,获得了纯度超过95%的重组IGFBP-3融合蛋白。Western-blot结果表明在相应分子量处有一条特异性条带。细胞活性研究显示它对MCF-7细胞生长具有一定的抑制作用,且在体外具有与IGF-I结合的活性。cDNA for Insulin-like growth factor binding protein 3 was cloned and constructed a prokaryotic expression vector pET-DsBA-IGFBP3. The construct was transformed into E. coli BL21 （DE3）plysS. The induced fusion protein（D-IGFBP3） was expressed successfully in soluble form. We obtained D-IGFBP3 the purify of which is over 95 % after purification by His affinity chromatography. The product was identified by Western-blot. The cell assay showed that the obtained fusion protein can inhibit the growth of MCF-7 and bind with IGF-I in vitro.

关 键 词：胰岛素样生长因子结合蛋白3(IGFBP-3) 高效可溶表达 纯化 生物学活性 

分 类 号：Q78[生物学—分子生物学]