| 大肠杆菌不耐热肠毒素B亚单位在大肠杆菌中的表达及其特性研究 |
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| 引用本文： | 葛俊伟,姜艳平,杨敏,哈卓,乔薪瑗,唐丽杰,李一经.大肠杆菌不耐热肠毒素B亚单位在大肠杆菌中的表达及其特性研究[J].东北农业大学学报,2010,41(6). |
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| 作者姓名： | 葛俊伟 姜艳平 杨敏 哈卓 乔薪瑗 唐丽杰 李一经 |
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| 作者单位： | 东北农业大学动物医学学院,哈尔滨,150030 |
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| 基金项目： | 国家科技支撑计划项目 |
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| 摘 要： | 将大肠杆菌不耐热性肠毒素B亚单位(LTB)基因eltb克隆到表达载体pET-28a中,转化大肠杆菌BL21/DE3/plyss。采用SDS-PAGE和Western blot分析,结果表明,在大肠杆菌中高效表达了LTB蛋白,其产量约为细菌总蛋白的20%,GM1-ELISA方法鉴定目的重组蛋白可与GM1结合,具有潜在的佐剂活性;由纯化的重组蛋白免疫小鼠制备的抗血清效价为1:16000,并且可以特异性识别天然毒素。结果表明,所表达的LTB有良好的抗原性和佐剂活性,制备的抗血清具有良好的免疫活性,为下一步以LTB作为佐剂的黏膜疫苗研究提供基础材料。
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| 关 键 词： | LTB 表达 活性 |
Expression and characterization of LTB subunit of Escherichia coli in Prokaryocyte |
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| GE Junwei,JIANG Yanping,YANG Min,HA Zhuo,QIAO Xinyuan,TANG Lijie,LI Yijing.Expression and characterization of LTB subunit of Escherichia coli in Prokaryocyte[J].Journal of Northeast Agricultural University,2010,41(6). |
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| Authors: | GE Junwei JIANG Yanping YANG Min HA Zhuo QIAO Xinyuan TANG Lijie LI Yijing |
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| Abstract: | The E.coli heat-labile enterotoxin B subunit gene (eltb) was amplified by the method of PCR, and subcloned into the expressing vector pET-28a. The recombinant plasmid was then transformed into the compound cell BL21/DE3 (plyss). After induced with IPTG, these cells expressed the recombinant protein LTB, identified by SDS-PAGE and Western blot. The results showed that the anti-LTB sera was prepared by immunizing BALB/C mice with purified LTB, and then analyzed by immunoblotting assay. The LTB gene was highly expressed in E.coli, and the amount of the expressed recombinant protein was about 20% of total cells. The titer of the antiserum after the second booster immunization was 1: 16 000 as determined by indirect ELISA. The reasult showed that this antiserum had specific reaction with both wild type LT and the recombinant LTB expressed in E.coli. |
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| Keywords: | LTB |
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