截断型人酸性成纤维细胞生长因子的表达及活性鉴定

Expression and Activity Identification of Shortened Human Acidic Fibroblast Growth Factor

摘要: 目的:稳定且具有活性的截断型人酸性成纤维细胞生长因子(shaFGF)的构建、表达及纯化.方法:以含全长人酸性成纤维细胞生长因子cDNA的质粒pUC-haFGF为模板,设计一对引物,PCR扩增获得shaFGF cDNA,将其克隆到表达载体pET3c中,重组质粒转化大肠杆菌BL21(DE3),0.5 mmol/L IPTG诱导表达,通过离子交换和肝素亲和层析纯化目标蛋白,MTT法检测促分裂活性.结果:shaFGF的表达量约为25%,纯化的shaFGF的促分裂活性与haFGF标准品相当.结论:BL21(DE3)/pET3c表达系统能高效表达shaFGF,纯化得到的shaFGF可供下一步的药学研究.

Abstract: AIM: To construct,overexpress and purify a stable and active form of shortened human acidic fibroblast growth factor (shaFGF) in Escherichia coli METHOD: Using the plasmid pUC-haFGF containing full-length human acidic fibroblast growth factor cDNA as a