三株递呈猪流行性腹泻病毒S基因的减毒鼠伤寒沙门菌的生物学特性研究  

Study on the Biological Characteristics of Three Attenuated Salmonella typhimurium Strains Harboring Porcine Epidemic Diarrhea Virus S Gene

作　　者：梁恩涛 陈杰[1] 黄小波[1,2] 文心田[1,2] 曹三杰[1,2] 吴学婧 朱书权[1] 

机构地区：[1]四川农业大学动物医学院动物传染病与基因芯片实验室,雅安625014 [2]四川农业大学人兽共患病研究室与猪病防治研究中心,成都611130

出　　处：《畜牧兽医学报》2015年第2期279-287,共9页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31072144;30901084);国家公益性行业(农业)科研专项(20123056)

摘　　要：为了探究减毒沙门菌递呈不同长度猪流行性腹泻病毒(PEDV)S基因的可行性,本研究对3株携带不同长度S基因片段的减毒鼠伤寒沙门菌进行了生物学特性评估。主要进行了对所构建的重组减毒沙门菌菌株SL7207(pVAX-S499-650)(长456bp,编码S蛋白N末端499—650位氨基酸)、SL7207(pVAX-S499-789)(长873bp,编码S蛋白N末端499—789位氨基酸)和前期构建的菌株SL7207(pVAXD-S1-789)(长2 367bp,编码S蛋白N末端1—789位氨基酸)的生物学特性研究,包括生长曲线测定、携带质粒的体内/外稳定性、回肠组织内的转录、在小鼠体内动态增殖规律、小鼠口服的安全性等。结果表明:SL7207(pVAX-S499-650)、SL7207(pVAX-S499-789)、SL7207(pVAXDS1-789)3种口服菌株均具有较高的稳定性;携带的目的基因可在肠道组织内转录;菌株在小鼠肝、脾中增殖约5周后逐渐被机体清除;小鼠口服具有良好的安全性。本研究结果表明减毒鼠伤寒沙门菌SL7207可递呈不同长度的S基因抗原片段,为后续开展3种菌株的实际口服免疫效果比较研究奠定了基础。To explore the feasibility of attenuated Salmonella typhimurium as the carrier of porcine epidemic diarrhea virus(PEDV)Sgene,the biological characteristics of three recombinant attenuated Salmonellastrains harboring varying length of PEDVSgene were researched.The biological characteristics research of SL7207(pVAX-S499-650)(456bp,encoding 499-650 amino acid of N-termanal S protein,SL7207(pVAX-S499-789)(873bp,encoding 499-789 amino acid of N-termanal S protein),SL7207(pVAXD-S1-789)(2 367 bp,encoding 1-789 amino acid of N-termanal S protein)which harboring PEDVSgenes in varying length plasmid DNA showed:SL7207(pVAXS499-650),SL7207(pVAX-S499-789)and SL7207(pVAXD-S1-789)were steady in vivo/vitro;the target genes could be transcribed in the ileum tissue of mouse;recombinant strains could colonize in liverand spleen of mice and gradually eliminated by host;safety to BALB/c mouse at the dosage of 1×109 CFU by oral administration.All of this,demonstrating the attenuated S.typhimuriumcould be the carrier of PEDVSgene and laying the foundation for evaluating the immunogenicity of recombinant attenuated Salmonellastrains in piglet.

关 键 词：猪流行性腹泻病毒 S基因 减毒鼠伤寒沙门菌 口服 生物学特性 

分 类 号：S858.28[农业科学—临床兽医学]