虹鳟胰岛素样生长因子ⅰ和ⅱ在大肠杆菌中的融合表达及促有丝分裂 .pdfVIP

动物学报 　49 (2) :266～271 , 2003 A cta Zoologica S inica 　　　　　 　 虹鳟胰岛素样生长因子 Ⅰ和 Ⅱ在大肠杆菌中的 融合表达及促有丝分裂活性 张为民 　张利红 ( 中山大学生命科学学院, 广州 510275) Fusion expression of rainbow trout ( Oncorhyncus mykiss) insulinlike growth factors Ⅰand Ⅱand their mitogenic activities in E . coli ZHAN G WeiMin 　ZHAN G LiHong ( ) School of L if e Sciences , Zhongshan U niversity , Guangz hou 　510275 , Chi na ( ) ( ) Abstract 　The preparation of recombinant rainbow trout insulinlike growth factors Ⅰ IGF Ⅰ and Ⅱ IGF Ⅱ using the His6 fusion polypeptide technique and the mitogenic activities of these compounds were investigated. Rainbow trout IGF Ⅰand IGF ⅡBCAD domain cDNA were subcloned into expression vector p ET15b (Novagen , USA) and ex pressed in host E . coli BL21 (DE3) as fusion polypeptides containing a stretch of 6 histidines at the Nterminus (referred as His6 fusion polypeptide) . The addition of IPTG (0 4 mmol/ L) induced the expression of polypeptides with a molecular weight of about 100 kDa. The expressed proteins were ammonium sulfate fractionally precipitated and further purified using a Ni2 + His ·Bind Resin (Novagen , USA) affinity column. The reduced SDSPA GE showed that IGF Ⅰwas of 80 % purit y and IGF Ⅱwas of 90 % purit y. The rainbow trout IGF ⅠHis6 fusion polypeptide showed dosedependent mitogenic effe