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泉古菌Sulfolobus islandicus PCNA1的原核表达及其多克隆抗体的制备
引用本文:田 斌,张昌毅,佘群新,梁运祥.泉古菌Sulfolobus islandicus PCNA1的原核表达及其多克隆抗体的制备[J].华中农业大学学报,2010,29(1):59-62.
作者姓名:田 斌  张昌毅  佘群新  梁运祥
作者单位:华中农业大学生命科学技术学院/农业微生物学国家重点实验室,武汉,430070
基金项目:科技部“973”计划项目(2003CB716004)资助
摘    要:泉古菌Sulfolobus islandicus中PCNA有3个同源类似蛋白,本研究选取PCNA1作为研究对象,为获得可溶性表达的PCNA1蛋白,制备多克隆抗体,深入了解PCNA1基因的功能,将PCNA1基因片段克隆到组氨酸标签融合的表达载体pET30a中,利用IPTG诱导,金属螯合亲和层析进行纯化分析表明,融合蛋白大部分可溶。紫外分光光度法测定纯化蛋白的纯度可达90%以上,浓度约为2 mg/mL。免疫家兔制备多克隆抗体,ELISA检测抗血清效价可达1∶10 000以上,Western印迹检测证明抗体特异性良好。

关 键 词:泉古菌  PCNA1基因  原核表达  金属螯合亲和层析  多克隆抗体  
收稿时间:2009/3/12 0:00:00
修稿时间:2009/6/23 0:00:00

Prokaryotic Expression of PCNA1from Crenarchaeaon Sulfolobus islandicus and Preparation of Its Polyclonal Antibody
TIAN Bin,ZHANG Chang-yi,SHE Qun-xin,LIANG Yun-xiang.Prokaryotic Expression of PCNA1from Crenarchaeaon Sulfolobus islandicus and Preparation of Its Polyclonal Antibody[J].Journal of Huazhong Agricultural University,2010,29(1):59-62.
Authors:TIAN Bin  ZHANG Chang-yi  SHE Qun-xin  LIANG Yun-xiang
Affiliation:TIAN Bin ZHANG Chang-yi SHE Qun-xin LIANG Yun-xiang College of Life Science , Technology/State Key Laboratory of Agricultural Microbiology,Huazhong Agricultural University,Wuhan 430070,China
Abstract:Based on the result of sequence analysis,there are three homologues of PCNA in Crenarchaeaon Sulfolobus islandicus and PCNA1 was selected as the subject of the current study.To obtain soluble protein of PCNA1 and its polyclonal antibody for studying the function of PCNA1 gene,the fragment of PCNA1 gene was amplified and cloned into the expression vector pET30a.Induced by IPTG,soluble protein of interest could be obtained.The protein was purified by metal chelate affinity chromatography and its purity and co...
Keywords:Sulfolobus islandicus  PCNA1 gene  prokaryotic expression  metal chelate affinity chromatography  polyclonal antibody  
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