%0 Journal Article
%T Construction, expression and Purification of a Novel Human Hepatic Growth Factor: Hepatopoietin Cn and Research on its Biological Activity in vitro<br>新的肝细胞生长因子HPPCn的表达、纯化及体外活性检测
%A WEI Ping
%A DU Shao-jun
%A CUI Chun-ping
%A ZHANG Da-jin
%A ZHANG Xiao-xiang
%A WU Zu-ze
%A <br>魏苹
%A 杜劭君
%A 崔春萍
%A 张达矜
%A 张霄翔
%A 吴祖泽
%J 中国生物工程杂志
%D 2007
%I 
%X Objective:To construct,express and purify recombinant human HPPCn protein and detect its biological acitivity in vitro.Methods:The prokaryotic expression vector pET-24a( )-HPPCn was constucted by template pUC18-HPPCn,and then was transformed into E.coli BL21,inducing it with 0.4mmol/L IPTG for 4 hours.The expression and solubility of recombinant protein was detected by SDS-PAGE.The interest protein was purified by anion exchange chromatography and its immunogenicity was identitied by Western blot analysis.Its biological activity in vitro was detected by MTT and 3H-TdR assays.Results:The rhHPPCn was highly expressed after IPTG inducing,and 23% of HPPCn was resoluble in thalline.The purity of HPPCn protein reached over 94% after twice anion exchange chromatography,and the Western blot analysis showed it could bind specifically to rabbit-HPPCn-polyclonal antibody.The MTT and 3H-TdR assays showed that the recombinant protein could remarkably promote proliferation of SMMC7721.Conclusion:rhHPPCn is successfully constructed and expressed in E.coli.The interest protein is successfully purified by twice anion exchange chromatography and is detected to promote the proliferation of SMMC7721.
%K HPPCn<br>原核表达
%K 阴离子交换层析
%K 肝癌细胞系增殖
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=272792C939874198A494A7B348650F2C&yid=A732AF04DDA03BB3&vid=DB817633AA4F79B9&iid=F3090AE9B60B7ED1&sid=B31275AF3241DB2D&eid=708DD6B15D2464E8&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=0&reference_num=17