布鲁氏菌毒力缺失疫苗株的构建

发布时间：2018-03-23 02:02

本文选题：基因工程　切入点：布鲁氏菌　出处：《基因组学与应用生物学》2017年01期 　论文类型：期刊论文

【摘要】：本研究在S19弱毒菌株的基础上通过一系列手段获得布鲁氏菌Bp26蛋白、Bmp18蛋白双缺失株,待遗传学鉴定合格后用于小鼠实验。将120只小鼠分为实验组、S19对照组和空白对照组3组,将双基因缺失株和S19弱毒株培养24 h后离心收集菌体,使用PBS清洗重悬后进行细菌计数,实验组小鼠腹腔接种检验合格的双基因缺失株5×10~5 CFU的剂量,病菌对照组腹腔接种S19弱毒菌株5×10~5 CFU的剂量,空白对照组腹腔注射同等体积的PBS溶液,观察分析Bp26蛋白做抗原的免疫原性、双基因缺失菌株的毒力情况和免疫应答情况。最终构建出能够区分自然免疫和人工免疫并具低毒性的布鲁氏弱毒菌株。双基因缺失株具有遗传稳定性,采用Bp26蛋白做抗原对3组小鼠的免疫原性测定,表明实验组40只小鼠均未表现出免疫原性,S19对照组40只小鼠中有28只表现出了免疫原性,占70%,空白对照组40只小鼠均未表现出免疫原性,双基因缺失株与原S19弱毒株的免疫应答基本一致,且更为安全。
[Abstract]:On the basis of S19 attenuated strain, the double deletion strain of Bp26 protein and Bmp18 protein of Brucella spp was obtained by a series of methods, and was used in mouse experiment after genetic identification. 120 mice were divided into three groups: the experimental group, S19 control group, and the blank control group, respectively. The double gene deletion strain and S19 attenuated strain were cultured for 24 hours and then centrifuged to collect the bacteria. The bacteria were cleaned by PBS and counted. The mice in the experimental group were inoculated intraperitoneally with the dose of 5 脳 10 5 CFU. The control group was intraperitoneally inoculated with S19 attenuated strain 5 脳 10 ~ (5) CFU, and the blank control group was intraperitoneally injected with the same volume of PBS solution. The immunogenicity of Bp26 protein as antigen was observed and analyzed. The virulence and immune response of the double gene deletion strain. Finally, a low toxicity Brucella attenuated strain, which can distinguish natural and artificial immunity, was constructed. The double gene deletion strain has genetic stability. The immunogenicity of the three groups of mice was determined by using Bp26 protein as antigen. The results showed that none of the 40 mice in the experimental group showed immunogenicity, and 28 of the 40 mice in the control group showed immunogenicity. None of the 40 mice in the blank control group showed immunogenicity, and the immune response of the double gene deletion strain was basically consistent with that of the original S19 attenuated strain, and it was safer than that of the original S19 attenuated strain.
【作者单位】：内蒙古民族大学动物科技学院;吉林大学动物科学学院;
【基金】：国家自然科学基金项目(31260608) 内蒙古自治区高等学校科学技术研究重点项目(NJZZ12117) 内蒙古通辽市校科技合作项目(SXZD2012131) 内蒙古自治区自然科学基金项目(2015MS0339)共同资助
【分类号】：S852.4

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