Wu Chen, Yao Guang-Yin, Zou Min-Ji, Chen Guang-Yu, Wang Min, Wang Jia-Xi, Xu Dong-Gang
Institute of Basic Medical Sciences, Beijing 100850, China.
Sheng Wu Gong Cheng Xue Bao. 2007 May;23(3):398-402.
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.
克隆胰岛素样生长因子结合蛋白3的cDNA，并构建原核表达载体——pET-DsBA-IGFBP3。将构建体转化到大肠杆菌BL21(DE3)plysS中。诱导的融合蛋白（D-IGFBP3）以可溶性形式成功表达。通过His亲和层析纯化后，我们获得了纯度超过95%的D-IGFBP3。通过蛋白质免疫印迹法对产物进行鉴定。细胞实验表明，所获得的融合蛋白在体外可抑制MCF-7细胞的生长并与胰岛素样生长因子-I（IGF-I）结合。
