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Expression and characterization of duck enteritis virus gI gene
Lijuan Li1, Anchun Cheng, Mingshu Wang
1Institute of Preventive Veterinary Medicine, Sichuan Agricultural University, Wenjiang, Chengdu city, Sichuan, 611130, PR China.
Researchers successfully expressed the duck enteritis virus (DEV) gI protein in E. coli, confirming its antigenicity and providing insights into its transcription and localization. This work aids in understanding DEV pathogenesis.
Area of Science:
- Virology
- Molecular Biology
- Protein Expression
Background:
- Alphaherpesvirus gI gene and protein roles in cell-to-cell spread are known.
- Limited information exists on the duck enteritis virus (DEV) gI gene characteristics.
Purpose of the Study:
- To express and characterize the DEV gI protein.
- To investigate the transcription and intracellular localization of DEV gI.
Main Methods:
- Cloning DEV gI ORF into pET-32a(+) vector for prokaryotic expression.
- Overexpression in E. coli BL21(DE3) induced by IPTG.
- Analysis by SDS-PAGE, PCR, sequencing, antibody generation, real-time PCR, and indirect immunofluorescence.
Main Results:
- Successfully constructed the pET-32a(+)-gI recombinant vector.
- Expressed a ~61 kDa His6-tagged gI protein in E. coli.
- Confirmed protein antigenicity, late-phase transcription, and cytoplasmic localization in infected cells.
Conclusions:
- DEV gI gene expression vector was successfully constructed.
- The expressed DEV gI protein retains antigenicity.
- Basic transcription and localization data provide clues for further functional analysis of DEV gI.
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