表达猫衣原体momp蛋白的重组狂犬病病毒SRV9-MOMP的构建与鉴定  

Construction and identification of recombinant rabies virus SRV9 strains expressing Chlamydia felis momp

作　　者：阮晓凤 齐宇 张艳艳 陈腾 周鑫涛 张守峰 扈荣良[1,2] RUAN Xiao-feng;QI Yu;ZHANG Yan-yan;CHEN Teng;ZHOU Xin-tao;ZHANG Shou-feng;HU Rong-liang(College of Animal Science and Technology,Jilin Agricultural University,Changchun 130118,China;Military Veterinary Institute,Academy of Military Medical Sciences,Academy of Military Sciences,Changchun 130122,China)

机构地区：[1]吉林农业大学动物科学技术学院,吉林长春130118 [2]军事科学院军事医学研究院军事兽医研究所,吉林长春130122

出　　处：《中国兽医科学》2019年第7期904-909,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31472176)

摘　　要：为构建表达猫衣原体主要外膜蛋白(momp)的重组狂犬病病毒,以狂犬病病毒SRV9株反向遗传质粒pUC57-SRV9为骨架,在P/M基因间隔序列处插入MOMP基因,获得正确的重组全长转录质粒pUC57-SRV9-MOMP(P/M)。再将全基因组质粒与pUC57-N(SRV9)、pUC57-P(SRV9)、pUC57-G(SRV9)、pUC57-L(SRV9)4个辅助质粒共转染BSR-T7细胞系。经直接免疫荧光法验证,成功拯救出病毒SRV9-MOMP。Western-blot和RT-PCR检测表明,momp在重组病毒SRV9-MOMP感染的细胞内被稳定表达。本研究证明狂犬病病毒SRV9株具有作为载体表达外源蛋白的潜力,为研制高效、安全、经济的猫衣原体病疫苗奠定了基础。To construct recombinant rabies virus(RABV)expressing the main outer membrane protein(momp)of Chlamydia felis,the MOMP gene was inserted into untranslated regioin between P and M genes in the pUC57-SRV9 including the cDNA of the RABV-SRV9 genome.The recombinant plasmid was co-transfected into BSR-T7 cell line with the helper plasmids pUC57-N(SRV9),pUC57-P(SRV9),pUC57-G(SRV9),and pUC57-L(SRV9).The recombinant virus SRV9-MOMP was successfully rescued by direct immunofluorescence assay.Western-blot and RT-PCR analysis showed that momp was stably expressed in BSR-T7 cells infected with SRV9-MOMP.This study demonstrated that the RABV SRV9 strain can be used to develop recombinant vaccines as a vector expressing foreign proteins,which lays the foundation for the development of effective,safe and economical vaccine against C.felis infection.

关 键 词：猫衣原体 主要外膜蛋白 重组狂犬病病毒 

分 类 号：S852.659.5[农业科学—基础兽医学]