| 大肠杆菌不耐热肠毒素B亚单位基因的原核表达 |
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| 引用本文： | 梁望旺,周丹娜,刘泽文,伍锐,杨克礼,段正赢,邓均华,徐涤平.大肠杆菌不耐热肠毒素B亚单位基因的原核表达[J].湖北农业科学,2009,48(11). |
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| 作者姓名： | 梁望旺 周丹娜 刘泽文 伍锐 杨克礼 段正赢 邓均华 徐涤平 |
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| 作者单位： | 湖北省农业科学院畜牧兽医研究所,武汉,430064 |
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| 基金项目： | 国家科技支撑计划项目，湖北省农业科技创新中心资助项目 |
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| 摘 要： | 大肠杆菌(Escherichia coli)不耐热肠毒素B亚单位(LTB)不仅是重要的黏膜免疫原,而且作为佐剂在黏膜免疫中起着重要的作用.试验采用PCR方法扩增了肠毒素性大肠杆菌的LTB基因,并将其克隆到pGEX-KG原核表达栽体上,构建了重组质粒pKG-LTB,经酶切和测序鉴定后,转化到大肠杆菌BL21(DE3)进行IPTG诱导表达,经SDS-PAGE和Western-blot分析表明,克隆的LTB基因与谷胱甘肽转移酶(GST)基因获得了高效融合表达,表达的融合蛋白GST-LTB分子量约为35kDa,并具有免疫反应活性.这为进一步研究LTB蛋白黏膜免疫原性和黏膜佐剂的作用奠定了基础.
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| 关 键 词： | 肠毒素大肠杆菌 不耐热肠毒素 克隆 原核表达 |
Prokaryotic Expression of the Escherichia coli Heat-labile Enterotoxin B Subunit Gene |
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| LIANG Wang-wang,ZHOU Dan-na,LIU Ze-wen,WU Rui,YANG Ke-li,DUAN Zheng-ying,DENG Jun-hua,XU Di-ping.Prokaryotic Expression of the Escherichia coli Heat-labile Enterotoxin B Subunit Gene[J].Hubei Agricultural Sciences,2009,48(11). |
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| Authors: | LIANG Wang-wang ZHOU Dan-na LIU Ze-wen WU Rui YANG Ke-li DUAN Zheng-ying DENG Jun-hua XU Di-ping |
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| Abstract: | The Escherichia coli heat-labile enterotoxin B subunit (LTB) was one of the most powerful mucosal immunogens and mucosal adjuvants. In this study, the LTB gene was cloned from enterotoxigenic E. coil and was fused with CST to construct prokaryotic expression plasmid pKG-LTB. After induction by IPTG, a high expression of fusion protein was obtained. SDS-PAGE analysis and Western blot showed that the fusion protein was 35kDa and the protein was specific to anti-CTB polyclonal antibody. The research laid a foundation for the further investigation of mucosal immunogenicity and mucosal adjuvant effect of protein LTB. |
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| Keywords: | Enterotoxigenic Escherichia coli heat-labile enterotoxin (LT) cloning prokaryotic expression |
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