摘要
大肠杆菌中表达的rhHGF α以包涵体形式存在 ,表达量约占菌体总蛋白的 2 5 %。为了建立一条简便的分离纯化生产工艺 ,我们研究了包涵体溶解及复性条件 ,并进行了活性测定。结果表明 ,用含有 4mol/L尿素的缓冲液洗涤包涵体 ,8mol/L尿素溶解包涵体后的上清 ,经过SephacrylS 2 0 0凝胶过滤、复性后 ,得到纯度达 90 %的rhHGF α ,SDS PAGE测定相对分子质量为 5 2 0 0 0。与rhHGF β体外共复性后 ,完整的rhHGF具有明显刺激原代培养大鼠肝细胞生长的活性。
Recombinant human hepatocyte growth f actor α chain produces as inclusion body in E.coli DH5α with 25% expressio n level.The solubilization and renaturation conditions of rhHGF α were studied to develop a way to facilitate the purification of inclusion body.Highly purifi e d inclusion body was obtained via several steps including washing with 4 mol/L u rea buffers,solubilization in 8 mol/L urea and Sephacryl S 200 chromatography.R elative m olecular weight of the rhHGF α determined by SDS PAGE was 52 000.After ren aturation with rhHGF β,the whole HGF shows biological activity of increasing t he growth of rat hepatocyte in primary culture.
出处
《山西医科大学学报》
CAS
2001年第2期104-106,共3页
Journal of Shanxi Medical University
基金
山西省自然科学基金资助项目!( 2 0 0 0 10 62 )
