title: Construction of an infectious plasmid clone of Muscovy duck parvovirus by TA cloning and creation of a partially attenuated strain.
abstract: 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.
authors.affiliation: a Graduate Institute of Microbiology and Public Health , National Chung Hsing University , Taichung , Taiwan.
authors.forename: T-Y
authors.initials: TY
authors.lastname: Yen
authors.forename: K-P
authors.initials: KP
authors.lastname: Li
authors.forename: S-C
authors.initials: SC
authors.lastname: Ou
authors.forename: J-H
authors.initials: JH
authors.lastname: Shien
authors.forename: H-M
authors.initials: HM
authors.lastname: Lu
authors.forename: P-C
authors.initials: PC
authors.lastname: Chang
mesh_terms: D000818:Animals; D003001:Cloning, Molecular / Q000379:methods; D017931:DNA Primers / Q000235:genetics; D004372:Ducks / Q000821:virology*; D016679:Genome, Viral / Q000235:genetics*; D017992:Parvovirus / Q000235:genetics* / Q000472:pathogenicity; D010957:Plasmids / Q000096:biosynthesis / Q000235:genetics*; D060888:Real-Time Polymerase Chain Reaction; D014162:Transfection; D014774:Virulence
chemical_list: D017931:DNA Primers