Rapid Construction of Recombinant PDCoV Expressing
an Enhanced Green Fluorescent Protein for the Antiviral Screening
Assay Based on Transformation-Associated Recombination Cloning in
Yeast
posted on 2024-01-05, 20:49authored byYucan Zhang, Chenxi Shi, Jin Yuan, Yue Zhang, Xiaohui Jin, Shaopo Zu, Honglei Zhang, Hui Hu
Porcine
deltacoronavirus (PDCoV) is an emerging enteropathogenic
coronavirus that mainly causes diarrhea and death in suckling piglets
and also has the potential for cross-species transmission, threatening
public health. However, there is still no effective vaccine or drug
to prevent PDCoV infection. In order to accelerate the development
of antiviral drugs, we established a high-throughput screening platform
using a novel genome editing technology called transformation-associated
recombination cloning in yeast. The recombinant PDCoV and PDCoV reporter
virus expressing enhanced green fluorescent protein were both rapidly
rescued with stable genealogical characteristics during passage. Further
study demonstrated that the reporter virus can be used for high-throughput
screening of antiviral drugs with a Z-factor of 0.821–0.826.
Then, a medicine food homology compound library was applied, and we
found that three compounds were potential antiviral reagents. In summary,
we have established a fast and efficient reverse genetic system of
PDCoV, providing a powerful platform for the research of antiviral
drugs.