中国动物传染病学报2024，Vol.32Issue(6)：182-186,5.
基于T7RNA聚合酶的犬瘟热病毒微型基因组构建与拯救
Construction and Rescue of the Minigenome of Canine Distemper Virus Based on T7 Promoter Expression System
摘要
Abstract
To develop a reverse genetic system for canine distemper virus(CDV)and recover the minigenome of CDV3 strain,the genome 3'-leader sequence,5'-tailer sequence and enhanced green fluorescent protein(EGFP)gene sequence were synthesized based on the genomic sequence of the commercial attenuated live vaccine CDV3 strain and reversely cloned into the plasmid PCR script regulated by the T7 promoter.Hammerhead ribozyme and hepatitis D ribozyme sequences were respectively introduced at the 5'and 3'ends of the minigenome to construct a minigenome plasmid pCR-mCDV3-EGFP.The pCR-mCDV3-EGFP plasmid and three helper plasmids expressing measles virus(MeV)N,P and L proteins were co-transfected into BSR cells and EGFP fluorescence was observed by fluorescence microscope.After 48 hours of co-transfection with plasmids,specific green fluorescence was observed in BSR cells.The successful development of a reverse genetics system of CDV minigenome was expected to be used for recovering CDV in the later period This study laid the foundation for the development of a new CDV vaccine and study of pathogenesis of infection.关键词
犬瘟热病毒/微型基因组/病毒反向遗传技术Key words
Canine distemper virus/minigenome/reverse genetics分类
农业科技引用本文复制引用
王慧,孙艺洋,王哲,赵建军..基于T7RNA聚合酶的犬瘟热病毒微型基因组构建与拯救[J].中国动物传染病学报,2024,32(6):182-186,5.基金项目
国家重点研发计划项目(2023YFD1800700) （2023YFD1800700）
黑龙江省大学生省级创新训练项目(s202310223041) （s202310223041）
