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

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38.5kD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。

Glasser'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-OmpP2re-combinant Lactococcus lactis.The protein expression and characterization of the recombinant bacteria were then performed.The results indicate that the OmpP2 protein has advantageous Band T cell epitopes which are potential candidate antigen.The recombinant Lactobacillus was successfully detected a target band with the size of approximately 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.