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Development of an improved reverse genetics system for avian metapneumovirus (aMPV): A novel vaccine vector protects against aMPV and infectious bursal disease virus

文献类型： 外文期刊

作者： Meng, Lingzhai 1 ; Chen, Yuntong 1 ; Yu, Mengmeng 1 ; Liu, Peng 1 ; Qi, Xiaole 1 ; Xue, Xiaoxiao 1 ; Guo, Ru 1 ; Zhang, Tao 1 ; Hu, Mingxue 1 ; Fan, Wenrui 1 ; Wang, Ying 1 ; Wang, Suyan 1 ; Zhang, Yanping 1 ; Liu, Yongzhen 1 ; Duan, Yulu 1 ; Cui, Hongyu 1 ; Gao, Yulong 1 ;

作者机构： 1.Chinese Acad Agr Sci, Avian Immunosuppress Dis Div, State Key Lab Anim Dis Control & Prevent, Harbin Vet Res Inst, Harbin 150069, Peoples R China

2.Natl Poultry Lab Anim Resource Ctr, Harbin 150069, Peoples R China

关键词： avian metapneumovirus; reverse genetics system; insertion site; protection; vector vaccines

期刊名称：JOURNAL OF INTEGRATIVE AGRICULTURE （ 影响因子：4.4； 五年影响因子：4.8 ）

ISSN： 2095-3119

年卷期： 2025 年 24 卷 5 期

页码：

收录情况： SCI

摘要： Avian metapneumovirus (aMPV), a paramyxovirus, causes acute respiratory diseases in turkeys and swollen head syndrome in chickens. This study established a reverse genetics system for aMPV subtype B LN16-A strain based on T7 RNA polymerase. Full-length cDNA of the LN16-A strain was constructed by assembling 5 cDNA fragments between the T7 promoter and hepatitis delta virus ribozyme. Transfection of this plasmid, along with the supporting plasmids encoding the N, P, M2-1, and L proteins of LN16-A into BSR-T7/5 cells, resulted in the recovery of aMPV subtype B. To identify an effective insertion site, the enhanced green fluorescent protein (EGFP) gene was inserted into different sites of the LN16-A genome to generate recombinant LN16-As. The results showed that the expression levels of EGFP at the site between the G and L genes of LN16-A were significantly higher than those at the other two sites (between the leader and N genes or replacing the SH gene). To verify the availability of the site between G and L for foreign gene expression, the VP2 gene of very virulent infectious bursal disease virus (vvIBDV) was inserted into this site, and recombinant LN16-A (rLN16A-vvVP2) was successfully rescued. Single immunization of specificprotection against the virulent aMPV subtype B and vvIBDV. Establishing a reverse genetics system here provides an important foundation for understanding aMPV pathogenesis and developing novel vector vaccines.

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