多聚胞嘧啶结合蛋白PCBP1在轮状病毒复制中的作用研究  

Study on the role of poly (C)-binding protein 1 in rotavirus replication

作　　者：纳璐 唐娅娅 周艳[1] 鲁辰星 刘军营 肖琳 李鸿钧[1] NA Lu;TANG Ya-ya;ZHOU Yan;LU Chen-xing;LIU Jun-ying;XIAO Lin;LI Hong-jun(Institute of Medical Biology,Chinese Academy of Medical Science and Peking Union Medical College,Kunming,Yunnan 650018,China)

机构地区：[1]中国医学科学院,北京协和医学院医学生物学研究所,云南省重大传染病疫苗研发重点实验室,云南昆明650018

出　　处：《中国病毒病杂志》2022年第1期15-20,共6页Chinese Journal of Viral Diseases

基　　金：国家自然科学基金(31700154);中央高校基本科研业务费项目(3332019109);云南省科技计划项目(2016FB034,2018ZF006);云南省科技厅科技计划项目(2019FB020);中国医学科学院医学与健康科技创新工程协同创新团队项目(2016-I2M-3-026)。

摘　　要：目的初步研究多聚胞嘧啶结合蛋白[poly (C)-binding protein, PCBP]1在轮状病毒复制过程中的作用和调控机制。方法构建携带有PCBP1基因的真核表达质粒pcDNA3.1-PCBP1,合成针对PCBP1的siRNA。通过转染pcDNA3.1-PCBP1和筛选出能够下调PCBP1基因的siRNA后,在MA104细胞中分别上调和下调PCBP1的表达,通过免疫荧光分析PCBP1对轮状病毒复制的作用。结果成功构建pcDNA3.1-PCBP真核表达质粒并且能在MA104细胞中过表达,通过免疫荧光发现上调PCBP1的表达水平后能够促进轮状病毒蛋白的表达,下调PCBP1后,病毒的复制减弱。结论初步证明PCBP1可促进轮状病毒复制,其具体的作用机制还有待进一步研究。Objective To study the role and regulatory mechanism of poly(C)-binding protein(PCBP) 1 in rotavirus replication. Methods The eukaryotic expression plasmid PCDNA3.1-PCBP1 carrying PCBP1 gene was constructed and the siRNA targeting PCBP1 was synthesized. After transfection of PCDNA3.1-PCBP1 and screening of siRNAs that can down-regulate PCBP1 gene, the expression of PCBP1 was up-regulated and down-regulated in MA104 cells, and the effect of PCBP1 on rotavirus replication was analyzed by immunofluorescence. Results The eukaryotic expression plasmid PCDNA3.1-PCBP was successfully constructed and overexpressed in MA104 cells. Immunofluorescence showed that upregulation of PCBP1 could promote the expression of rotavirus protein, while down-regulation of PCBP1 could weaken the replication of the virus. Conclusion PCBP1 promotes rotavirus replication, and the specific mechanism remains to be further studied.

关 键 词：多聚胞嘧啶结合蛋白(PCBP1) 轮状病毒 真核表达质粒 siRNA 病毒复制 

分 类 号：R373.2[医药卫生—病原生物学]