%0 Journal Article
%T Prokaryotic expression and preparation of polyclonal antibody for human SUMO-3 gene<br>人SUMO-3基因原核表达及多克隆抗体制备
%A Ning AN
%A <br>安宁
%A 杨振
%A 宋振
%A 雷鸣
%A 郭泽坤
%J 中国生物工程杂志
%D 2008
%I 
%X To construct a prokaryotic expression vector of Human SUMO-3, purify Human SUMO-3 proteins produced by the expression system, and prepare its antiserum. The fragment of human SUMO-3 gene was amplified from pEYPF-SUMO-3 plasmid by PCR. The enzyme digested target fragment was cloned into pET41a(+) clone and expression vector and transfected into E. coli. BL 21 (DE3) plysS, in which SUMO-3 expression was induced by IPTG. After the soluble protein was purified through GST affinity chromatography, processed by identified by SDS-PAGE, a rabbit was immunized with the fusion protein, and the antiserum was obtained. The result of DNA sequence analysis showed that the cloned SUMO-3 gene sequence was completely corresponding to GenBank data. SDS-PAGE and Western blotting showed that the expressed SUMO-3 fusion protein was about 44.0 kDa, mainly existing soluble protein of E. coli. , that could be purified through GST affinity chromatography. The results of ELISA is positive, and Western blotting confirmed that the antiserum reacted specifically to the SUMO-3 protein. A recombinant SUMO-3 protein and the specific polyclonal antibody have been obtained, which provides a basis for establishment of immunoassays of human SUMO-3.
%K Human SUMO-3
%K prokaryotic expression
%K protein purification
%K polyclonal antibody<br>人SUMO-3
%K 原核表达
%K 蛋白纯化
%K 多克隆抗体
%K 基因原核表达
%K 多克隆
%K 抗体制备
%K Gene
%K Human
%K Polyclonal
%K Antibody
%K Preparation
%K Expression
%K 用工
%K 功能
%K 家族
%K SUMO
%K 研究
%K 实验
%K 抗体效价
%K 阳性
%K 检测显示
%K ELISA
%K 特异性识别
%K Western
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=951380788B355DEA7D75520FA3B199C9&aid=22EE3BE271730FA9BF7F423932F9487C&yid=67289AFF6305E306&vid=D3E34374A0D77D7F&iid=94C357A881DFC066&sid=117F81797AB182FC&eid=08805F9252973BA4&journal_id=1671-8135&journal_name=中国生物工程杂志&referenced_num=0&reference_num=15