[1]赵宁,王芳,恽时锋,等.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,(08):648-652.
 ZHAO Ning,WANG Fang*,YUN Shi-feng,et al.The expression and immunoprotection of the truncated dermonecrotic toxin of rabbit Bordetella bronchiseptica[J].Chinese journal of preventive veterinary medicine,2011,(08):648-652.
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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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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年08期
页码:
648-652
栏目:
免疫学
出版日期:
2011-08-15

文章信息/Info

Title:
The expression and immunoprotection of the truncated dermonecrotic toxin of rabbit Bordetella bronchiseptica
作者:
赵宁;王芳;恽时锋;范志宇;胡波;
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;南京军区南京总医院比较医学科;
Author(s):
ZHAO Ning1WANG Fang1*YUN Shi-feng2FAN Zhi-yu1HU 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
关键词:
支气管败血波氏杆菌坏死毒素原核表达免疫保护
Keywords:
Bordetella bronchiseptica dermonecrotic toxin prokaryotic expression immunoprotection
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。
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 with the recombinant proteins,and all the mice in immunized groups were survived when challenged intraperitoneally with virulent B.bronchiseptic strain BJL0504 at 2 dosages of LD50,while the mice mortility were 60% in control group.Furthermore,the bacteria in lung was eliminated completely from the immunned mice post challenging intranasally with B.bronchiseptic(1.07×108 cfu/mL) at 9 days,on the contrary,the bactera counts in mice lung were up to 1.2×105 cfu in control group.These results shown that the DNT1 and DNT3 had strong immunoprotection against B.bronchiseptic infection and could be used as a subunit vaccine against B.bronchiseptic

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备注/Memo

备注/Memo:
基金: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]
更新日期/Last Update: 2011-09-29