title: A comparison of AMPV subtypes A and B full genomes, gene transcripts and proteins led to reverse-genetics systems rescuing both subtypes.
abstract: Avian metapneumovirus (AMPV) infection of poultry causes serious disease in most countries and subtype A reverse-genetic (RG) systems have allowed a generation of viruses of known sequence, and proved useful in developments towards better control by live vaccines. While subtype B viruses are more prevalent, bacterial cloning issues made subtype B RG systems difficult to establish. A molecular comparison of subtype A and B viruses was undertaken to assess whether subtype A RG components could be partially or fully substituted. AMPV subtype A and B gene-end sequences leading to polyadenylation are, to our knowledge, reported for the first time, as well as several leader and trailer sequences. After comparing these alongside previously reported gene starts and protein sequences, it was concluded that subtype B genome copies would be most likely rescued by a subtype A support system, and this assertion was supported when individual subtype A components were successfully substituted. Application of an advanced cloning plasmid permitted eventual completion of a fully subtype B RG system, and proved that all subtype-specific components could be freely exchanged between A and B systems.
authors.affiliation: Department of Infection Biology, University of Liverpool, Leahurst Campus, Neston, Cheshire, CH64 7TE, UK.
authors.forename: Andrea
authors.initials: A
authors.lastname: Laconi
authors.affiliation: Department of Infection Biology, University of Liverpool, Leahurst Campus, Neston, Cheshire, CH64 7TE, UK.
authors.forename: Jayne
authors.initials: J
authors.lastname: Clubbe
authors.affiliation: Department of Infection Biology, University of Liverpool, Leahurst Campus, Neston, Cheshire, CH64 7TE, UK.
authors.forename: Marco
authors.initials: M
authors.lastname: Falchieri
authors.affiliation: Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra, 50, 40064, Ozzano Emilia, BO, Italy.
authors.forename: Caterina
authors.initials: C
authors.lastname: Lupini
authors.affiliation: Department of Animal Medicine, Production and Health, University of Padua, Viale dell'università, 16, Legnaro, PD, Italy.
authors.forename: Mattia
authors.initials: M
authors.lastname: Cecchinato
authors.affiliation: Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra, 50, 40064, Ozzano Emilia, BO, Italy.
authors.forename: Elena
authors.initials: E
authors.lastname: Catelli
authors.affiliation: Department of Veterinary Medical Sciences, University of Bologna, Via Tolara di Sopra, 50, 40064, Ozzano Emilia, BO, Italy.
authors.forename: Valeria
authors.initials: V
authors.lastname: Listorti
authors.affiliation: Department of Infection Biology, University of Liverpool, Leahurst Campus, Neston, Cheshire, CH64 7TE, UK.
authors.forename: Clive J
authors.initials: CJ
authors.lastname: Naylor
mesh_terms: D003001:Cloning, Molecular; D015870:Gene Expression; D020869:Gene Expression Profiling; D016679:Genome, Viral; D005838:Genotype; D029121:Metapneumovirus / Q000235:genetics* / Q000502:physiology*; D059386:Reverse Genetics / Q000379:methods; D014764:Viral Proteins / Q000235:genetics*; D014779:Virus Replication
chemical_list: D014764:Viral Proteins