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Updated: Nov 29, 2025

Author Spotlight: Exploring Cloning Techniques for Full-Length DNA Fragments
Published on: May 17, 2024
Development and utilization of an infectious clone for porcine deltacoronavirus strain USA/IL/2014/026
Xufang Deng1, Alexandra C Buckley2, Angela Pillatzki3
1Department of Microbiology and Immunology, Loyola University Chicago, Stritch School of Medicine, Maywood, IL, 60153, USA.
Abstract:
We report the generation of a full-length infectious cDNA clone for porcine deltacoronavirus strain USA/IL/2014/026. Similar to the parental strain, the infectious clone virus (icPDCoV) replicated efficiently in cell culture and caused mild clinical symptoms in piglets. To investigate putative viral interferon (IFN) antagonists, we generated two mutant viruses: a nonstructural protein 15 mutant virus that encodes a catalytically-inactive endoribonuclease (icEnUmut), and an accessory gene NS6-deletion virus in which the NS6 gene was replaced with the mNeonGreen sequence (icDelNS6/nG). By infecting PK1 cells with these recombinant PDCoVs, we found that icDelNS6/nG elicited similar levels of type I IFN responses as icPDCoV, however icEnUmut stimulated robust type I IFN responses, demonstrating that the deltacoronavirus endoribonuclease, but not NS6, functions as an IFN antagonist in PK1 cells. Collectively, the construction of a full-length infectious clone and the identification of an IFN-antagonistic endoribonuclease will aid in the development of live-attenuated deltacoronavirus vaccines.

