%0 Journal Article
%T Expression, Purification and Renaturation of ProNGF in Escherichia coli<br>ProNGF在大肠杆菌中的表达、纯化和复性
%A Hanmin Jiang
%A Xinjun Chai
%A Bing He
%A Juan Zhao
%A Xinda Yu
%A <br>江汉民
%A 柴新君
%A 何冰
%A 赵娟
%A 俞新大
%J 生物工程学报
%D 2008
%I 
%X Nerve growth factor (NGF) promotes neuronal survival and differentiation and stimulates neurite outgrowth. NGF is synthesized as a precursor-proNGF in vivo. In this paper, a pET-proNGF prokaryocyte expression vector was constructed and transformed into E. coli BL21(DE3)pLysS. The proNGF was expressed in the form of non-active aggregated monomer in E. coli after induction with IPTG. SDS-PAGE revealed the proNGF expression product had a Mr.30.2 kD. Western blotting analysis showed that the protein had good antigenicity. Fusion protein was successfully purified by Ni2+-NTA affinity chromatography and cleaved by Enterokinase and 13.1 mg proNGF was obtained from 100 mL cell culture in a typical experiment. The protein was dialyzed in a redox system containing reduced and oxidized glutathione. RP-HPLC was used to analysis the result of the refolding. The refolded proNGF protein can induce neurite outgrowth of PC12 cells, which indicated that pro-form of NGF we obtained had biological activity.
%K nerve growth factor
%K expression
%K propeptide
%K protein refolding
%K biological activity<br>神经生长因子
%K 表达
%K 前导肽
%K 蛋白复性
%K 生物活性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=E4461B0FDF34F1DA6E8FBA1FB8905CCF&yid=67289AFF6305E306&vid=B91E8C6D6FE990DB&iid=38B194292C032A66&sid=4ECB3941871FD391&eid=C4BBAD7A2DCC89BC&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=13