摘要
目的 对人工合成的人广谱细胞生长因子基因进行体外表达与纯化 ,并测定其活性。方法 将人广谱细胞生长因子基因克隆于 pGEX 4T 1表达载体 ,转化于大肠杆菌BL2 1菌株进行诱导表达 ,利用亲和层析纯化表达产物 ,通过体外培养的大鼠成纤维细胞存活实验检测其活性。结果 携带重组质粒的菌株经诱导产生高水平的表达产物 ,纯化的表达产物具备较高的纯度 ,并基本保有其天然活性。结论 广谱细胞生长因子表达体系的成功建立 。
Objective To express the basic fibroblast growth factor (bFGF) gene in bacteria, to purify the product and to determine its activity.Methods The bFGF gene was cloned into pGEX 4T 1 expression vector. BL21 strain of Escherichia coli was transformed with the recombinant vector and induced to express bFGF protein. The protein was purified by affinity chromatography. The biological activity of purified protein was estimated by the survival of cultured rat fibrobasts.Results The BL21 strain of Escherichia coli with recombinant vector showed high level of bFGF gene expression after induction. The product was purified successfully and maintained its natural activity.Conclusion The expression and purification of recombinant bFGF with natural activity facilitate its research and application in future.
出处
《同济大学学报（医学版）》
CAS
2001年第5期7-9,共3页
Journal of Tongji University(Medical Science)
基金
铁道部医学专项基金资助项目 (B9972 )
