%0 Journal Article
%T Use of Ssp dnaB Mini-intein as a Fusion Partner for Preparation of Recombinant Human Brain Natriuretic Peptide<br>Ssp dnaB蛋白质内含子介导的重组人脑钠素的制备
%A DI Lie
%A ZHANG Hong-Wei
%A XU Lang-Lai
%A <br>狄洌
%A 张宏伟
%A 徐朗莱
%J 生物工程学报
%D 2006
%I 
%X Human brain natriuretic peptide (hBNP) was used clinically for the treatment of acute decompensated congestive heart failure. In this paper, hBNP was expressed as a fusion protein with a histidine tag and Ssp dnaB mini-intein which was capable of self-cleavage. After affinity chromatography with Ni-Sepharose and renaturation, the fusion protein was enriched with CM-cellulose. Ssp dnaB mini-intein mediated peptide-bond hydrolysis was triggered by shifting the pH and temperature in the CM-cellulose column, which let to the release of hBNP from the fusion protein and the separation of hBNP from His-DnaB. The hBNP sample was further purified by C4 reverse phase HPLC, and 2.8mg of the peptide with homogeneity of 97% was obtained from one liter of culture medium. The biological activity was assayed in vitro, which indicated that hBNP had a potent vasodilatory effect on rabbit aortic strips with an EC50 of 1.94 x 10(-6) mg/mL.
%K polypeptide preparation
%K brain natriuretic peptide
%K intein
%K protein splicing<br>多肽制备
%K 脑钠素
%K 蛋白质内含子
%K 蛋白质剪接
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=0B8E9AA2E913CEE1&yid=37904DC365DD7266&vid=BC12EA701C895178&iid=0B39A22176CE99FB&sid=F1A8654ADB4E656E&eid=50BBDFAC8381694B&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=3&reference_num=17