| 兔支气管败血波氏杆菌DNT蛋白的分段表达及其免疫保护性研究 |
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| 引用本文： | 赵宁,王芳,恽时锋,范志宇,胡波. 兔支气管败血波氏杆菌DNT蛋白的分段表达及其免疫保护性研究[J]. 中国预防兽医学报, 2011, 33(8). DOI: 10.3969/j.issn.1008-0589.2011.08.16 |
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| 作者姓名： | 赵宁 王芳 恽时锋 范志宇 胡波 |
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| 作者单位： | 1. 江苏省农业科学院兽医研究所 农业部动物疫病免疫与诊断重点开放实验室,国家兽用生物制品工程技术研究中心,江苏南京210014
2. 南京军区南京总医院比较医学科,江苏南京,210002 |
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| 基金项目： | 现代农业产业技术体系建设专项资金(nycytx-44); 江苏省农业科技自主创新资金[CX(10)214] |
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| 摘 要： | 皮肤坏死毒素(DNT)是支气管败血波氏杆菌(Bb)的主要毒力因子之一。为研究DNT结合位点和催化位点的免疫保护性,本研究分别将其核苷酸序列N-端的1 612 bp片段(DNT1)和C-端的973 bp片段(DNT3)克隆到原核表达载体pET-28a(+)中,经IPTG诱导后在E.coli BL21(DE3)中获得表达。SDS-PAGE和western blot检测表明重组蛋白DNT1、DNT3均具有良好的反应原性。在主动免疫保护试验中,重组蛋白免疫组小鼠均能够产生较高的DNT抗体水平;当使用2 LD50的Bb强毒株BJL0504进行腹腔攻毒后,其存活率为100%,而阴性对照组则为40%;当使用1.07×108 cfu/mL Bb强毒株BJL0504进行滴鼻攻毒后第9 d时,重组蛋白免疫组小鼠已基本清除肺脏内的Bb菌,而阴性对照组小鼠肺脏内的Bb菌落数仍高达1.2×105 cfu/肺。本实验结果表明,DNT的结合位点和催化位点均具有良好的免疫保护性,有希望作为疫苗添加成分,为新型疫苗的研制提供实验依据。
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| 关 键 词： | 支气管败血波氏杆菌 坏死毒素 原核表达 免疫保护 |
The expression and immunoprotection of the truncated dermonecrotic toxin of rabbit Bordetella bronchiseptica |
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| ZHAO Ning,WANG Fang,YUN Shi-feng,FAN Zhi-yu,HU Bo. The expression and immunoprotection of the truncated dermonecrotic toxin of rabbit Bordetella bronchiseptica[J]. Chinese Journal of Preventive Veterinary Medicine, 2011, 33(8). DOI: 10.3969/j.issn.1008-0589.2011.08.16 |
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| Authors: | ZHAO Ning WANG Fang YUN Shi-feng FAN Zhi-yu HU Bo |
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| Affiliation: | ZHAO Ning1,WANG Fang1*,YUN Shi-feng2,FAN Zhi-yu1,HU Bo1(1.Ministry of Agricultural National Center for Engineering Research of Veterinary Bio-products,Key Laboratory of Animal Diseases Diagnostic and Immunology,Institute of Veterinary Medicine,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China,2.Department of Comparative Medicine,Nanjing General Hospital of Nanjing Command,Nanjing 210002,China) |
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| Abstract: | Dermonecrotic toxin(DNT) is identified as one of the main virulence factor of Bordetella bronchiseptica.To study the immunoprotection of the binding and catalytic regions of DNT,the N-terminal 1,612 bp(DNT1) and the C-terminal 973 bp(DNT3) fragments of the DNT were cloned into the pET-28a(+) and expressed in E.coli BL21(DE3),respectively.The expression of recombinant protein DNT1 and DNT3 were identified by SDS-PAGE and western blot analysis.High level antibody against DNT was induced in the mice immunized ... |
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| Keywords: | Bordetella bronchiseptica dermonecrotic toxin prokaryotic expression immunoprotection |
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