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Open Access 01-12-2017 | Research

Preliminary study of the UL55 gene based on infectious Chinese virulent duck enteritis virus bacterial artificial chromosome clone

Authors: Ying Wu, Yangguang Li, Mingshu Wang, Kunfeng Sun, Renyong Jia, Shun Chen, Dekang Zhu, Mafeng Liu, Qiao Yang, Xinxin Zhao, Xiaoyue Chen, Anchun Cheng

Published in: Virology Journal | Issue 1/2017

Abstract

Background

Lethal Duck Enteritis Virus (DEV) infection can cause high morbidity and mortality of many species of waterfowl within the order Anseriformes. However, little is known about the function of viral genes including the conserved UL55 gene among alpha herpes virus due to the obstacles in maintenance and manipulation of DEV genome in host cells.

Methods

In this paper, we constructed an infectious bacteria artificial chromosome (BAC) clone of the lethal clinical isolate duck enteritis virus Chinese virulent strain (DEV CHv) by inserting a transfer vector containing BAC mini-F sequence and selection marker EGFP into UL23 gene using homologous recombination. UL55 deletion and its revertant mutant were generated by two-step RED recombination in E. coli on basis of rescued recombinant virus. The function of UL55 gene in DEV replication and its effect on distribution of UL26.5 protein were carried out by growth characteristics and co-localization analysis.

Results

The complete genome of DEV CHv can be stably maintained in E. coli as a BAC clone and reconstituted again in DEF cells. The generated UL55 deletion mutant based on DEV CHv-BAC-G displayed similar growth curves, plaque morphology and virus titer of its parental virus in infected Duck Embryo Fibroblast (DEF) cells. Immunofluorescence assay indicated that the loss of UL55 gene do not affect the distribution of UL26.5 protein in intracellular. These data also suggest infectious BAC clone of DEV CHv will facilitate the gene function studies of DEV genome.

Conclusions

We have successfully developed an infectious BAC clone of lethal clinical isolate DEV CHv for the first time. The generated UL55 gene mutant based on that demonstrated this platform would be a very useful tool for functional study of DEV genes. We found the least known DEV UL55 is dispensable for virus replication and UL26.5 distribution, and it could be a very promise candidate locus for developing bivalent vaccine. Experiment are now in progress for testifying the possibility of UL55 gene locus as an exogenous gene insertion site for developing DEV vectored vaccine.

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Title: Preliminary study of the UL55 gene based on infectious Chinese virulent duck enteritis virus bacterial artificial chromosome clone
Authors: Ying Wu
Yangguang Li
Mingshu Wang
Kunfeng Sun
Renyong Jia
Shun Chen
Dekang Zhu
Mafeng Liu
Qiao Yang
Xinxin Zhao
Xiaoyue Chen
Anchun Cheng
Publication date: 01-12-2017
Publisher: BioMed Central
Published in: Virology Journal / Issue 1/2017
Electronic ISSN: 1743-422X
DOI: https://doi.org/10.1186/s12985-017-0748-y

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Preliminary study of the UL55 gene based on infectious Chinese virulent duck enteritis virus bacterial artificial chromosome clone

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Methods

Results

Conclusions

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Abstract

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Conclusions

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