Construction and rescue of Muscovy duck-origin goose parvovirus from an infectious clone containing an E-box deletion within the left terminal region

• Published in: Molecular and cellular probes 2018-12, Vol.42, p.32-35
• Main Authors: Wang, Shao, Xiao, Shifeng, Cheng, Xiaoxia, Chen, Shaoying, Zhu, Xiaoli, Lin, Fengqiang, Chen, Shilong
• Format: Article
• Language: English
• Subjects: E-box motif; Goose parvovirus; Infectious clone; Muscovy duck
• ISSN: 0890-8508; 1096-1194
• DOI: 10.1016/j.mcp.2018.09.003

To obtain a deletion mutant of Muscovy duck-origin goose parvovirus (MDGPV) and to analyze its biological characteristics, the pMDGPVPT plasmid, which contains a full-length DNA infectious clone of the MDGPV PT strain, was used in this study as the template. The E-box at nt 315 of the left inverted terminal repeat sequence (L-ITR) was deleted by overlap extension PCR to obtain the infectious recombinant plasmid p-PTΔE315. The p-PTΔE315 plasmid was transfected into 9-day-old non-immune Muscovy duck embryos via the yolk sac and the rescued deletion mutant virus r-PTΔE315 was generated. Experiments to demonstrate the novel deletion mutant virus’ biological characteristics showed that r-PTΔE315 can cause typical lesions after infection of Muscovy duck embryos. Compared with its parent strain PT, the virulence of r-PTΔE315 and its proliferation ability in Muscovy duck embryos were attenuated, but its ability to replicate in MDEF cells was enhanced. This study laid the foundation for further understanding of the relationship between E-box deletion in the L-ITR and MDGPV virulence. •In this study, we constructed a full-length infectious clone (pMDGPVPT).•Deletion of E-box (E315) may improve the adaptability of MDGPV to MDEF cells.•Deletion of E-box (E315) decreased MDGPV replication in Muscovy duck embryos.