%0 Journal Article %T Prokaryotic expression and preparation of polyclonal antibody for human SUMO-3 gene
人SUMO-3基因原核表达及多克隆抗体制备 %A Ning AN %A
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
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