生物技术通报2025，Vol.41Issue(12)：95-105,11.DOI:10.13560/j.cnki.biotech.bull.1985.2025-0496
中国丁型流感病毒株D/JY3002反向遗传操作系统的构建与功能验证
Establishment and Functional Validation of a Reverse Genetics System for the Chinese Influenza D Virus D/JY3002
摘要
Abstract
[Objective]The current prevalent influenza D virus in Chinese cattle herds primarily belongs to the D/Yama2019 genetic lineage.The development of a reverse genetics system for the D/Yama2019 genetic lineage influenza D virus offers a crucial research tool for the in-depth investigation of its replication and pathogenic mechanisms.[Method]The Chinese influenza D virus strain D/bovine/CHN/JY3002/2022(Abbreviated as D/JY3002),belonging to the D/Yama2019 genetic lineage,was utilized as the research material.The DNA fragments corresponding to the genomic segments that encode the primary antigen of the virus,Hemagglutinin-Esterase-Fusion(HEF),were successfully cloned into the engineered bidirectional expression vector pCC1-DualPro.The DNA fragments corresponding to the remaining genomic segments were seamlessly cloned into the widely utilized bidirectional expression vector pHW2000.Subsequently,the recombinant influenza D virus was successfully rescued through improved and optimized operational procedures.[Result]The rescued influenza D virus strain(rD/JY3002)demonstrated a replication ability comparable to that of the naturally isolated Chinese influenza D virus strain(D/JY3002),and the replication kinetics of the two viruses are similar.Moreover,the rD/JY3002 virus can be stably propagated for at least five generations.Using the established reverse genetics system,recombinant influenza D viruses containing the green fluorescence reporter gene(GFP)and the HEF genes from other genetic lineages(D/OK and D/660)were successfully generated.They were respectively designated as rD/JY3002-GFP,rD/JY3002-D/OK-HEF,and rD/JY3002-D/660-HEF.These results confirmed that reassortment occurred among influenza D viruses from different genetic lineages.[Conclusion]An efficient and stable reverse genetics system for the influenza D virus Chinese strain has been established.This system can be utilized to further advance exogenous gene presentation technology,employing the influenza D virus as a vector.关键词
丁型流感病毒/反向遗传操作系统/基因组节段/载体Key words
influenza D virus/reverse genetics system/genomic segment/vector引用本文复制引用
HU Wan-ke,YU Jie-shi,CHEN Yun-xia,LUO Di-zhou,WU Si-yu,LI Jian-bo,ZHAI Shao-lun,JU Xiang-hong,LIAO Ming,WEI Wen-kang..中国丁型流感病毒株D/JY3002反向遗传操作系统的构建与功能验证[J].生物技术通报,2025,41(12):95-105,11.基金项目
国家自然科学基金青年项目(32202795),广东省自然科学基金面上项目(2024A1515011049),广东省农业科学院"中青年学科带头人""金颖之星"培养项目(R2023PY-JX025),广东省农业科学院人才专项(青年科技骨干)项目(R2021YJ-QG008) （32202795）
