养猪2024,Issue(2) :41-47.

副猪嗜血杆菌OmpP2基因分子特征及重组乳酸菌的构建与鉴定

Molecular characteristics of OmpP2 gene in Glaesserella parasuis and construction and identification of its recombinant lactic acid bacteria

韩敬娴 周媛媛 贾永超 韩凯龙 刘子蔚 尹荣兰 金悦 尹荣焕
养猪2024,Issue(2) :41-47.

副猪嗜血杆菌OmpP2基因分子特征及重组乳酸菌的构建与鉴定

Molecular characteristics of OmpP2 gene in Glaesserella parasuis and construction and identification of its recombinant lactic acid bacteria

韩敬娴 1周媛媛 1贾永超 1韩凯龙 1刘子蔚 1尹荣兰 2金悦 1尹荣焕1
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作者信息

  • 1. 重要家畜疫病研究教育部重点实验室/沈阳农业大学动物科学与医学学院,辽宁 沈阳 110866
  • 2. 吉林省畜牧兽医科学研究院,吉林 长春 130062
  • 折叠

摘要

猪格拉瑟病是由副猪嗜血杆菌引起的猪的主要细菌性传染病.构建安全、有效的新型疫苗是防控该病的研究热点.本研究对OmpP2 基因编码的蛋白质进行生物信息学预测分析后,应用无缝克隆技术把PCR扩增获得的OmpP2 基因连接到pNZ8148 载体,构建出pNZ8148-OmpP2重组载体,转入乳酸乳球菌感受态细胞,获得pNZ8148-OmpP2 重组乳酸乳球菌;再进行蛋白表达与鉴定.结果表明:生物信息学预测到OmpP2 蛋白具备优势B细胞和T细胞表位,是具有潜力的候选抗原;经重组乳酸乳球菌蛋白表达与鉴定,检测到大小约为38.5 kD的目的条带,与预期大小相符.说明乳酸乳球菌能作为递送载体成功表达G.parasuis异源蛋白.本实验为后续G.parasuis乳酸菌活载体疫苗的研制奠定了基础.

Abstract

Glässer's disease is a major bacterial disease affecting the development of pig farming.Constructing a novel vaccine that can safely and effectively control the disease has been a hot topic in epidemic prevention and control research.In this study,following bioinformatics prediction and analysis,we constructed the pNZ8148-OmpP2 recombinant vector by ligating the OmpP2 gene into the pNZ8148 vector utilizing seamless cloning technolo-gy.Then transfected the recombinant vector into Lactococcus lactis receptor cells to obtain pNZ8148-OmpP2 re-combinant Lactococcus lactis.The protein expression and characterization of the recombinant bacteria were then performed.The results indicate that the OmpP2 protein has advantageous B and T cell epitopes which are potential candidate antigen.The recombinant Lactobacillus was successfully detected a target band with the size of approxi-mately 38.5 kD,which was consistent with the expected size,indicating that Lactococcus lactis could be used as a delivery vector to express G.parasuis heterologous protein successfully.Our research laid the foundation for the subsequent development of the live recombinant Lactococcus lactis vaccine for G.parasuis.

关键词

副猪嗜血杆菌/OmpP2基因/乳酸乳球菌/活载体疫苗

Key words

Glaesserella parasuis/OmpP2 gene/Lactococcus lactis/Live vector vaccine

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基金项目

国家自然科学基金(32172861)

辽宁省自然科学基金(2021-MS-221)

辽宁省教育厅科研项目(LSNJC201917)

出版年

2024
养猪
东北养猪研究会

养猪

影响因子:0.395
ISSN:1002-1957
参考文献量1
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