Position: Home > Articles > Construction and Expression of eae-stx1/2B Fusion Gene from EHEC O157∶H7
Chinese Journal of Veterinary Science
2005,25
(3)
270-272
出血性大肠杆菌O157 eae-stx1/2B融合基因的构建和表达
作 者：
陈萍;刘军;祝令伟;孙洋;郭学军;齐翀;冯书章;郑明光
单 位：
解放军军事医学科学院军事兽医研究所
关键词：
EHEC O157∶H7;紧密素;志贺毒素;融合基因
摘 要：
构建表达eae和stx1/2B的融合基因,克隆eae基因的C端280个氨基酸残基(Int280)基因部分,以正确地阅读框定向插入到含有stx1/2B融合基因的质粒,构建重组质粒,将其转化于BL21(DE3),用IPTG进行诱导表达,经SDS-PAGE电泳检测,该融合蛋白获得了高效表达。薄层扫描分析表明目的蛋白表达量占菌体总蛋白含量的50.67%。由于该融合蛋白由eae、stx1B、stx2B等三部分抗原组成,可刺激机体产生针对紧密素和StxB的抗体,在E-HECO157亚单位疫苗设计或单克隆抗体抗制备中具有重要价值。
译 名：
Construction and Expression of eae-stx1/2B Fusion Gene from EHEC O157∶H7
作 者：
CHEN Ping,LIU Jun,ZHU Ling-wei,SUN Yang,GUO Xue-jun,QI Chong,FENG Shu-zhang*, ZHENG Ming-guang (The Military Veterinary Institute,Academy of Military Medical Sciences of PLA,Changchun 130062,China)
关键词：
EHEC O157∶H7;intimin;Stx;fusion gene
摘 要：
To construct eae-stx1/2B fusion gene,the DNA fragment encoding C-terminal 280 amino acids(Int280) of intimin was fused in frame with the stx1/2B gene.We created a recombinant plasmid by insertion of eae-stx1B-stx2B fusion gene into pET-28a,the recombinant plasmid was transformed into BL21(DE_3),and the fusion protein was expressed in E.coli BL21 efficiently with IPTG.Analysis of SDS-PAGE and thin layer scanning results showed that amount of expressed fusion protein was 50.67% in total of bacteria protein.Because the fusion protein was consisted of eae Int280,stx1B and stx2B antigens,it can evoke immune responses against Int280,Stx1B and Stx2B.The eae-stx1/2B fusion protein expressed in E.coli could offer an important ingredient that should be consider in construction of recombinant EHEC O157 vaccine or preparation of experimental material for the monoclonal antibody.
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