摘要
红斑丹毒丝菌血清型较多,目前商品化疫苗主要为活疫苗和灭活疫苗,血清型为1型和2型,尚无商品化亚单位疫苗。该研究旨在研究一种具有广谱免疫效果的重组蛋白。从临床分离株SerF5中获得的spaA基因,经密码子优化和截短后在spaA N端融合鼠伤寒沙门氏菌的鞭毛蛋白(nfliC),构建大肠埃希氏菌BL21(DE3)重组表达质粒,通过诱导nfliC-SpaA重组蛋白表达和镍柱亲和层析纯化后,分别在小鼠和仔猪上进行免疫效果试验。结果显示,获得的nfliC-SpaA重组蛋白在大肠埃希氏菌中呈可溶性表达,纯化后重组蛋白具有良好的免疫原性,可在小鼠和仔猪体内产生较高的体液免疫抗体水平,且对1型和2型红斑丹毒丝菌菌株混合攻毒的小鼠和仔猪均可提供完全保护。说明获得的nfliC-SpaA重组蛋白具有良好的免疫效果,为猪丹毒亚单位疫苗的开发提供了技术支持。
There are many serotypes of Erysipelothrix rhusiopathiae,and the commercial vaccines are mainly alive and inactivated,with serotype 1 and serotype 2,and there is no commercial subunit vaccine.This study aimed to develop a recombinant protein with broad-spectrum immune effects.In this study,SpaA gene was obtained from clinical isolate SerF5.After codon optimization and truncation,flagellin(nfliC)of Salmonella typhimurium was fused to the N-terminal of SpaA to construct recombinant expression plasmid of Escherichia coli BL21(DE3).The expressed recombinant protein was induced and purified by nickel column affinity chromatography,The immunogenicity analysis was studied in mice and piglets.The results showed that the obtained nfliC-SpaA with adjuvant and immune-enhancing effects was soluble,and the purified recombinant protein had good immunogenicity,and could produce high humoral immune antibody levels in mice and piglets,and could provide complete protection against the mixed challenge of Erysipelothrix rhusiopathiae strains serotype 1 and serotype 2.It can be seen that the nfliC-SpaA recombinant protein successfully obtained has good immune efficacy,which lays a strong technical support for the development of Erysipelothrix rhusiopathiae subunit vaccine.
作者
雷妍丽
严晨光
金云云
关洋
王孟月
王彦伟
黄玉欣
逄文强
田克恭
LEI Yan-li;YAN Chen-guang;JIN Yun-yun;GUAN Yang;WANG Meng-yue;WANG Yan-wei;HUANG Yu-xin;PANG Wen-qiang;TIAN Ke-gong(National Research Center For Veterinary Medicine,Luoyang,Hennan 471000,China;Pulike Biological Engineering,Inc.,Luoyang,Hennan 471000,China)
出处
《动物医学进展》
北大核心
2025年第4期22-27,共6页
Progress In Veterinary Medicine
基金
国家“十四五”重点研发计划项目(2023YFD1800505
2023YFD1800304-4)。
关键词
红斑丹毒丝菌
重组蛋白
免疫保护
佐剂
亚单位疫苗
Erysipelothrix rhusiopathiae
Recombinant protein
Immune protection
Adjuvant
Subunit vaccine
作者简介
雷妍丽(1993-),女,河南省洛阳市人,硕士研究生,工程师,主要从事蛋白表达与纯化研究,同等贡献作者;严晨光(1997-),男,河南省焦作市人,本科生,同等贡献作者;通讯作者:逄文强;通讯作者:田克恭。
