%0 Journal Article
%T Cloning and Expression of Urate Oxidase and Its Application in Serum Uric Acid Analysis<br>尿酸氧化酶基因的克隆、表达及其产物的应用
%A X J Zhu
%A J G Liu
%A G X Li
%A <br>朱献军
%A 刘建国
%A 黎高翔
%J 生物工程学报
%D 2001
%I 
%X Anurate oxidase (uricase, EC 1.7.3.3) gene from Candida utilis AS2.117 was cloned by PCR amplification with primers derived from conserved regions of published uicase DNA sequence. The DNA sequence of cloned uricase gene was determined and a high homology compared to the reported gene was found. The cloned gene was inserted into Bam H I and Nde I sites of pET21a to create the recombinant plasmid pURO. In Escherichia coli BL21(DE3) host, the expression lever of uricase reached to about 40% of total soluble proteins of the cell. The western blot analysis confirmed the result of expression. Properties of the enzyme protein produced by E. coli BL21(DE3)/pURO were determined and similar with those of original protein from Candida utilis AS2.117. Furthermore, the thermostability of the expressed protein was enhanced. The purified recombinant uricase was used in serum uric acid analysis.
%K Candida utilis
%K AS2
%K 117
%K uricase
%K gene cloning and expression
%K uric acid analysis<br>产朊假丝酵母，尿酸氧化酶，基因克隆和表达，尿酸分析
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=A92C1B2F9B0B3C9F&yid=14E7EF987E4155E6&vid=BCA2697F357F2001&iid=CA4FD0336C81A37A&sid=68D88C2FCF9C3098&eid=AA76E167F386B6B3&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=8&reference_num=15