马链球菌兽疫亚种PepO蛋白原核表达及其免疫保护试验  

Expression and immunogenicity of PepO protein of Streptococcus equis subspecies zooepidemicus

作　　者：孙莹 尚天甜 刘旭明 韩莹 摆淑玲 彭婕[1] SUN Ying;SHANG Tiantian;LIU Xuming;HAN Ying;BAI Shuling;PENG Jie(College of Veteri-nary Medicine,Gansu Agriculture University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物医学院,甘肃兰州730070

出　　处：《中国兽医学报》2022年第1期95-99,113,共6页Chinese Journal of Veterinary Science

基　　金：甘肃农业大学公招博士科研启动基金资助项目(GAU-KYQD-2018-26);甘肃农业大学省级大学生创新创业训练计划资助项目(202010733010);甘肃农业大学学生科研训练基金资助项目(SRTP)(202003014);兰州市人才创新创业基金资助项目(2019-RC-115)。

摘　　要：以马链球菌兽疫亚种(Streptococcus equi subsp.zooepidemicus,SEZ)ATCC35246株基因组为模板,根据NCBI数据库已公布的pepO基因序列设计引物进行扩增,利用同源重组技术连接至pCold-SUMO质粒进行原核表达及纯化,并以纯化的蛋白制备兔抗SEZ PepO多克隆抗体。将兔多克隆抗体与SEZ野生菌株混合后进行mBMEC细胞的被动抗体保护试验,同时以不加抗体组作为对照,结果显示PepO多抗能够在细菌黏附侵袭细胞的过程中对细胞形成有效保护。经纯化后蛋白免疫的BALB/c小鼠腹腔注射5×10^(4)CFUATCC35246菌液(5×LD_(50)),连续观察并记录7日内小鼠的临床表现及存活情况,结果显示PepO免疫组小鼠存活率达到70%。结果表明,SEZ PepO具有良好的免疫保护效力,能够作为SEZ亚单位疫苗的候选抗原。The pepO gene of Streptococcus equi subsp.zooepidemicus(SEZ)was amplified and ligated to pCold-SUMO plasmid using homologous recombination technology.After expression and purification,the purified protein was used to prepare rabbit anti-SEZ PepO polyclonal antibody.mBMEC cell adhesion and invasion test were carried out using rabbit polyclonal antibody mixed with SEZ wild strain.The group without antibody was set as the control.The results showed that PepO polyclonal antibody could effectively protect cells from being invaded.BALB/c mice immunized with purified protein were intraperitoneally injected with 5×10^(4)CFU ATCC35246 bacterial solution(5×LD_(50)).The clinical manifestation and survival of mice were observed and recorded within 7 d.The results showed that the survival rate of mice reached 70%in the PepO group.The results indicate that SEZ PepO has good immune protection efficacy and can be used as a candidate antigen for SEZ subunit vaccines.

关 键 词：马链球菌兽疫亚种 PepO 致病机制 免疫保护 

分 类 号：S852.4[农业科学—基础兽医学] S852.61[农业科学—兽医学]