中国兽医科学2025，Vol.55Issue(7)：870-876,7.DOI:10.16656/j.issn.1673-4696.2025.0113
宿主蛋白TRIM7对口蹄疫病毒复制的影响
Impact of host protein TRIM7 on the replication of foot-and-mouth disease virus
摘要
Abstract
This study aimed to explore the effect of TRIM7 on the replication of foot-and-mouth disease virus(FMDV).Two single guide RNAs(sgRNAs)targeting the exons of porcine TRIM7 gene were designed and were respectively constructed into the vector PX459.The successfully constructed sgRNA recombinant plasmids were transfected into PK-15 cells.Monoclonal cells were selected by puromycin screening and the limited dilution method.The PK-15 cell lines with the knockout of the TRIM7 gene were identified by sequencing and Western-blot analysis.Subsequently,the TRIM7 gene-knockout cells and control cells were infected with FMDV,respectively.The effect of the knockout of the TRIM7 gene on the replication of FMDV was comprehensively evaluated by Western-blot,RT-qPCR,and TCID50 determination.Meanwhile,TRIM7 was overexpressed in PK-15 cells to detect its effect on virus replication.The results showed that the PK-15 cell line with the knockout of the TRIM7 gene was successfully constructed,and the replication level of FMDV in the TRIM7 gene-knockout cell line was significantly lower than that in the control cells.When TRIM7 was overexpressed,it significantly promoted the replication of the virus,indicating that TRIM7 affected the replication of FMDV.This study laid the foundation for further exploring the biological functions of TRIM7 and the molecular mechanisms of its regulation of viral replication.关键词
CRISPR/Cas9/TRIM7基因/细胞系/过表达/口蹄疫病毒Key words
CRISPR/Cas9/TRIM7 gene/cell line/overexpression/foot-and-mouth disease virus分类
农业科技引用本文复制引用
刘泓屹,张桂红,张伟,蔡建涛,古涛鸣,王良锋,陈治彤,曹伟军,郑海学,杨帆..宿主蛋白TRIM7对口蹄疫病毒复制的影响[J].中国兽医科学,2025,55(7):870-876,7.基金项目
国家自然科学基金项目(32072831,32102639) （32072831,32102639）
国家重点研发计划项目(2021YFD1800300) （2021YFD1800300）
甘肃省杰出青年基金项目(21JR7RA026) （21JR7RA026）
中央高校基本科研业务费专项(lzujbky-2022-ey20) （lzujbky-2022-ey20）
甘肃省科技重大专项(22ZD6NA001) （22ZD6NA001）
