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Identification and characterization of the duck enteritis virus (DEV) US2 gene
1Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu, Sichuan, China.
Genetics and Molecular Research : GMR
|November 5, 2015
Summary
Duck enteritis virus US2 protein is a late gene, transcribed and translated after infection. This protein is packaged into virions and found in the cytoplasm and plasma membrane of infected cells.
Area of Science:
- Virology
- Molecular Biology
Background:
- The US2 protein's role in duck enteritis virus (DEV) infection is known, but its expression kinetics, cellular localization, and presence in virions were uncharacterized.
- Understanding these aspects is crucial for elucidating DEV pathogenesis and viral assembly.
Purpose of the Study:
- To investigate the expression kinetics and cellular localization of the DEV US2 protein during infection.
- To determine if the US2 protein is incorporated into DEV virions.
Main Methods:
- Gene amplification via PCR, recombinant protein expression, and polyclonal antibody generation.
- Quantitative real-time PCR, ganciclovir (GCV) and cycloheximide (CHX) treatment, Western blot, and indirect immunofluorescence assays.
- Protease protection assay to assess virion packaging.
Main Results:
- US2 gene transcription began at 24 hours post-infection (pi) and peaked at 72 hours pi.
- US2 protein expression was detected from 48 hours pi, inhibited by GCV or CHX, indicating it's a viral gene product.
- US2 protein localized to the plasma membrane and cytoplasm of infected cells and was packaged into DEV virions.
Conclusions:
- DEV US2 is a late viral gene, with its protein product potentially serving as a tegument component.
- The findings provide insights into DEV replication, virion structure, and the evolutionary conservation of US2 homologs in alphaherpesviruses.
