%0 Journal Article
%T Fermentation, Purification and Identification of Recombinant RGD-Hirudin<br>重组双功能水蛭素的发酵、纯化和鉴定
%A MO Wei
%A ZHANG Yan-Ling
%A WANG Long-Sheng
%A YANG Xin-Ying
%A SONG Hou-Yan
%A <br>莫炜
%A 张艳玲
%A 王龙生
%A 杨新英
%A 宋后燕
%J 生物工程学报
%D 2004
%I 
%X Recombinant RGD-Hirudin ( r-RGD-Hirudin ) has double functions: anti-thrombin activity and anti-platelet aggregation activity. To identify these functions, the expression plasmid, RGD-Hirudin-pPIC9K, was constructed by inserting cDNA of RGD-hirudin in yeast expression vector pPIC9K. The high expression clone was gained after screening. This clone was fermented for 3 days. The r-RGD-hirudin was secreted into the culture. It was ultra-filtrated from culture supernatant, then after gel filtration chromatography and anion exchange chromatography, the purified r-RGD-hirudin was gained. Its purity was larger than 97% and its specific activity was 12 000 ATU/mg. The yield per liter culture of purified r-RGD-hirudin was 1 g and overall recovery yield was more than 75%. The purified r-RGD-hirudin was identified by reductive SDS-PAGE, anti-thrombin activity assay, anti-platelet aggregation assay, LC/MS and isoelectrofocusing assay. It is proved that r-RGD-Hirudin is ramification of wt-Hirudin and it has anti-thrombin activity and anti-platelet aggregation activity .
%K r-RGD-hirudin
%K high expression
%K fermentation
%K purification
%K identification<br>重组双功能水蛭素(r-RGDHirudin)，
%K 高效表达，
%K 发酵，
%K 纯化，
%K 鉴定
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=1D35D3993D9603E6&yid=D0E58B75BFD8E51C&vid=A04140E723CB732E&iid=CA4FD0336C81A37A&sid=6FBD78E3BAB60869&eid=28F8B56DB6BEE30E&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=3&reference_num=11