A comparison of AMPV subtypes A and B full genomes, gene transcripts and proteins led to reverse-genetics systems rescuing both subtypes

Laconi, Andrea, Clubbe, Jayne, Falchieri, Marco, Lupini, Caterina, Cecchinato, Mattia, Catelli, Elena, Listorti, Valeria and Naylor, Clive J (2016) A comparison of AMPV subtypes A and B full genomes, gene transcripts and proteins led to reverse-genetics systems rescuing both subtypes. JOURNAL OF GENERAL VIROLOGY, 97 (6). pp. 1324-1332.

Text resubmission - JGV-D-16-00019_R1.pdf - Author Accepted Manuscript Download (2MB)

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.

Item Type:	Article
Uncontrolled Keywords:	Metapneumovirus, Viral Proteins, Cloning, Molecular, Gene Expression Profiling, Virus Replication, Gene Expression, Genotype, Genome, Viral, Reverse Genetics
Depositing User:	Symplectic Admin
Date Deposited:	18 May 2017 10:14
Last Modified:	08 Feb 2023 04:35
DOI:	10.1099/jgv.0.000450
Related URLs:	Author Publisher
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3007531