Construction of an infectious plasmid clone of Muscovy duck parvovirus by TA cloning and creation of a partially attenuated strain

• Published in: Avian pathology Vol. 44; no. 2; pp. 124 - 128
• Main Authors: Yen, T.-Y, Li, K.-P, Ou, S.-C, Shien, J.-H, Lu, H.-M, Chang, P.-C
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 04.03.2015; Taylor & Francis Ltd
• Subjects: Animal diseases; Animals; Cairina moschata; Cloning; Cloning, Molecular - methods; DNA Primers - genetics; duck eggs; ducklings; Ducks - virology; fibroblasts; genome; Genome, Viral - genetics; Muscovy; Muscovy duck parvovirus; Mutagenesis; Mutation; Parvovirus; Parvovirus - genetics; Parvovirus - pathogenicity; Plasmids; Plasmids - biosynthesis; Plasmids - genetics; Poultry; Real-Time Polymerase Chain Reaction; site-directed mutagenesis; Transfection; vaccine development; Virulence; Viruses
• ISSN: 1465-3338; 0307-9457; 1465-3338
• DOI: 10.1080/03079457.2015.1008399

Muscovy duck parvovirus (MDPV) infection is a highly contagious and fatal disease of Muscovy ducklings. The infectious clone methodology is a valuable tool to study the pathogenic mechanisms of viruses, but no infectious clone of MDPV is yet available. In this study, a plasmid clone containing the full-length genome of MDPV was constructed using the TA cloning methodology. This MDPV clone was found to be infectious after transfection of primary Muscovy duck embryo fibroblast cells and passage in embryonated Muscovy duck eggs. Site-directed mutagenesis showed that the K75N mutation in the VP1 protein of MDPV resulted in the partial attenuation of the virus. The availability of an MDPV infectious clone can facilitate investigation of the pathogenic mechanisms of MDPV and development of vaccines against diseases caused by MDPV.