畜牧与兽医2025，Vol.57Issue(6)：98-104,7.
猪TRIM56 E3泛素连接酶活性突变体的构建及功能研究
Construction and function of swine TRIM56 E3 ubiquitin ligase activity mutant
摘要
Abstract
This study was to construct a swine TRIM56 E3 ubiquitin ligase activity mutant and to verify its role in the STING-mediated type I interferon signaling pathway.Here,homology comparison and amino acid sequence analysis of swine and human TRIM56 were performed to identify the swine TRIM56 E3 ubiquitin ligase active site.Then,primers for the swine TRIM56 E3 ubiquitin ligase activity mutant were de-signed,the swine TRIM56-E3 ubiquitin ligase active sites were site-mutated,the swine TRIM56 E3 ubiquitin ligase activity mutant plasmid was constructed,and its expression was verified by Western blot.Finally,the role of swine TRIM56 and its E3 ubiquitin ligase activity mu-tant in the STING-mediated type Ⅰ IFN signaling pathway was explored by Luciferase assay,Real-time PCR assay and Co-IP assay.The results showed that the swine TRIM56 E3 ubiquitin ligase activity mutant plasmid was successfully constructed,that swine TRIM56 promoted the STING-mediated IFN-β mRNA level and promoter activity,and catalyzed the K63-linked polyubiquitination of STING;while the swine TRIM56 E3 ubiquitin ligase activity mutant had no significant effect.This study provided an effective tool for future research on the role of swine TRIM56 in antiviral natural immunity.关键词
猪TRIM56/E3泛素连接酶/定点突变/STINGKey words
swine TRIM56/E3 ubiquitin ligase/site-specific mutation/STING分类
农业科技引用本文复制引用
张金霞,周萌,王先炜,杜以军,曹莹,张昊文,刘德鹏,李铎,王晓冉,田似报,吴香菊,齐静..猪TRIM56 E3泛素连接酶活性突变体的构建及功能研究[J].畜牧与兽医,2025,57(6):98-104,7.基金项目
中央引导地方基金发展项目(YDZX2024019) （YDZX2024019）
山东省科技型中小企业创新能力提升项目(2024TSGC0274) （2024TSGC0274）
山东省农业科学院农业科技创新项目(CXGC2024D10,CXGC2025A04) （CXGC2024D10,CXGC2025A04）
