摘要
旨在筛选并鉴定与猪繁殖与呼吸综合征病毒(PRRSV)核衣壳蛋白(N蛋白)相互作用的宿主蛋白。本试验通过构建猪肺酵母cDNA文库、酵母双杂交、免疫共沉淀、激光共聚焦、过表达和干扰试验等手段,筛选与N蛋白互作的宿主蛋白并研究其对PRRSV复制的影响。结果显示:N蛋白和核糖体蛋白S20(RPS20)在酵母细胞内发生互作,免疫共沉淀和激光共聚焦试验进一步确定N蛋白和RPS20蛋白互作并共定位于细胞质中。过表达和干扰试验证实RPS20能够影响HP-PRRSV在细胞内的增殖。本研究丰富了PRRSV与宿主因子互作的网络,为进一步研究RPS20蛋白在PRRSV感染过程中的生物学功能奠定了基础。
The present study aimed to screen and identify host proteins that interact with the nucleocapsid protein(N protein) of pathogenic porcine reproductive and respiratory syndrome virus(PRRSV). In this experiment, we screened host proteins that interact with N protein and investigated their effects on PRRSV replication by constructing a porcine lung yeast cDNA library, yeast two-hybrid, immunoprecipitation, laser confocal, overexpression and interference assays. The results showed that N protein and ribosomal protein S20(RPS20) interacted with each other in yeast cells. The overexpression and interference assays also provided preliminary evidence that RPS20 can affect the proliferation of PRRSV in cells. This study enriches the network of PRRSV-host factor interactions and lays the foundation for further studies on the biological functions of RPS20 protein during PRRSV infection.
作者
王君君
宋慧敏
李倬伟
姜平
周双海
李焕荣
刘雪威
WANG Junjun;SONG Huimin;LI Zhuowei;JIANG Ping;ZHOU Shuanghai;LI Huanrong;LIU Xuewei(College of Animal Science and Technology Beijing University of Agriculture,Beijing 102206,China;College of Veterinary Medicine,Nanjing Agricultural University,Nanjing 210095,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2023年第1期293-303,共11页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
北京农学院青年教师科研创新能力提升计划(QJKC2022016)
国家自然科学青年科学基金项目(32102663)。
