[1]张艳,陈创夫,张辉,等.布鲁氏菌M5-90ΔWboA基因缺失株的构建及免疫效果的初步评价[J].中国预防兽医学报,2011,(07):546-551.
 ZHANG Yan,CHEN Chuang-fu,ZHANG Hui,et al.Construction and preliminary evaluation of immune effects of deletion mutant of WboA gene of Brucella melitensis vaccine M5-90[J].Chinese journal of preventive veterinary medicine,2011,(07):546-551.
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布鲁氏菌M5-90ΔWboA基因缺失株的构建及免疫效果的初步评价
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2011年07期
页码:
546-551
栏目:
免疫学
出版日期:
2011-07-15

文章信息/Info

Title:
Construction and preliminary evaluation of immune effects of deletion mutant of WboA gene of Brucella melitensis vaccine M5-90
作者:
张艳;陈创夫;张辉;张沾;李志强;张俊波;孟仁;王震;
石河子大学动物科技学院/新疆地方与民族高发病教育部重点实验室;
Author(s):
ZHANG Yan CHEN Chuang-fu ZHANG Hui ZHANG Zhan LI Zhi-qiang ZHANG Jun-bo MENG Ren WANG Zhen
Key Laboratory of Xinjiang Endemic and Ethnic Disease, College of Animal Science & Technology, Shihezi University, Shihezi 832003, China
关键词:
布鲁氏菌WboA 基因同源重组免疫原性
Keywords:
Brucella WboA gene homologous recombination immunogenicity
摘要:
为获得毒力较弱并能区分自然感染和疫苗免疫的布鲁氏菌候选疫苗株,本研究用PCR方法扩增WboA基因的上下游同源臂序列,构建重组质粒pGEM-7zf-Δ WboA-Sac,电转化布鲁氏菌M5-90感受态细胞,筛选布鲁氏菌疫苗株M5-90的WboA基因缺失株,并对获得的M5-90Δ WboA遗传稳定性、毒力、免疫保护性、抗体水平等指标进行检测。实验结果表明M5-90Δ WboA株的毒力比M5-90株明显减弱,差异极显著(p<0.01),体液免疫和细胞免疫结果表明M5-90Δ WboA株与亲本M5-90株相比差异不显著(p<0.05),M5-90Δ WboA株和亲本株的保护率分别为10%和20%,表明M5-90Δ WboA株与M5-90株具有相似的保护性。凝集试验和western blot试验显示M5-90Δ WboA株免疫小鼠的血清反应结果为阴性。本研究构建的布鲁氏菌基因缺失株M5-90Δ WboA具有较好的遗传稳定性,毒力比亲本株更弱,免疫保护性与亲本株相当,并能以血清学检测方法区分野毒株感染和缺失疫苗株免疫的动物。
Abstract:
Vaccination is a major measure for prevention of brucellosis, but it is unable to be distinguished from the vaccinated to natural infected animals. In this study, the WboA gene was knocked out of the genomic DNA of Brucella melitensis vaccine M5-90 strain to construct the recombinant M5-90Δ WboA by homologous recombination. The test results showed that M5-90Δ WboA was less virulent than that of the parent M5-90 strain(p<0.01). Humoral immunity and cellular immunity tests showed that there was no significant difference between M5-90Δ WboA and M5-90 parent strain (p<0.05). BALB/c mice were immunized with M5-90 Δ WboA or M5-90 and challenged with virulent strain 16M and the survival rate were 10% and 20%, respectively, indicating that the M5-90Δ WboA provided the similar protection of M5-90 strain. Agglutination test and western blot showed that the serum response of M5-90Δ WboA in vaccinated mice were negative. These results indicated that M5-90ΔWboA strain might be a promising vaccine against brucellosis, and could be distinguished from the vaccine immunization to natural infection in animals by serum test

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备注/Memo

备注/Memo:
基金:973计划项目(2010CB530203);;国家自然科学基金项目(30960288、31001046)
更新日期/Last Update: 2011-09-29