猪瘟病毒C株表位突变毒株的构建及拯救  

The Construction and Rescue of Epitope Mutant Strain of Classical Swine Fever Virus C Strain

作　　者：刘元杰 徐璐[1] 朱元源[1] 徐嫄[1] 张乾义[1] 李翠[1] 李明[1] 夏应菊 王琴[1] 刘业兵[1] 赵启祖[1] 邹兴启[1] LIU Yuanjie;XU Lu;ZHU Yuanyuan;XU Yuan;ZHANG Qianyi;LI Cui;LI Ming;XIA Yingju;WANG Qin;LIU Yebing;ZHAO Qizu;ZOU Xingqi(National Swine Fever Reference Laboratory of China Institute of Veterinary Drug Control,Beijing 100081,China)

机构地区：[1]中国兽医药品监察所国家猪瘟参考实验室,北京100081

出　　处：《畜牧兽医学报》2024年第2期698-705,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：中国兽医药品监察所“兽药行业公益性重点专项”-猪瘟疫苗免疫与自然感染(野毒感染)鉴别诊断技术研究。

摘　　要：本研究旨在突变猪瘟病毒C株WH303位点,构建感染性克隆,为猪瘟病毒C株标记疫苗的研究提供候选毒株。本试验以猪瘟病毒C株为研究材料,经重叠PCR方法突变WH303位点,再分别连接至C株和C-flag株全长感染性克隆。体外转录RNA,转染PK15细胞,细胞连续传代完成病毒的拯救和增殖。结果显示,经特异性限制内切酶酶切反应验证,感染性克隆构建成功。经RT-PCR、过氧化物酶单层细胞试验检测,表明病毒成功拯救。病毒细胞传代至17代,检测到病毒,表明病毒可以稳定传代。综上,本研究成功构建了C株WH303突变感染性克隆MC和MC-flag,并成功拯救MC株,得到可以稳定传代的病毒株。An infectious clone was constructed by mutating the WH303 site of CSFV C strain to provide candidate strains for the study of marker vaccine of CSFV C strain.In this study,CSFV C strain was used as the research material,and WH303 site was mutated by overlapping PCR method,and then cs-full and CS-full-flag were cloned into the full-length infectious clones of CSFV C strain.In vitro transcription RNA,and then transfected into PK15 cells,continuous passage of cells to.through the passage to complete the cell rescue and proliferation.results through The results showed that the specific restriction endonuclease digestion reaction verified the successful construction of infectious clone.verify the success of infectious clone construction RT-PCR and peroxidase monolayer cell test detection,indicating the success of the virus rescue indicated that the virus had been rescued successfully.The virus can still be detected in the 17th generation of virus cell,indicating that the virus can be stably passaged.In summary,MC and MC-flag of C strain WH303 mutated infectious clones were constructed successfully in this study,MC strain was saved successfully,and the stable virus strain was obtained.

关 键 词：猪瘟病毒 C株 WH303位点 突变 感染性克隆 

分 类 号：S852.651[农业科学—基础兽医学]