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Molecular cloning and characterization of the UL31 gene from Duck enteritis virus

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Abstract

Using a combination of bioinformation analysis and Dot blot technique, a gene, designated hereafter as the duck enteritis virus (DEV) UL31 gene (GenBank accession number EF643559), was identified from the DEV CHv genomic library. Then, the UL31 gene was cloned and sequenced, which was composed of 933 nucleotides encoding 310 amino acids. Multiple sequence alignment suggested that the UL31 gene was highly conserved in Alphaherpesvirinae and similar to the other herpesviral UL31. Phylogenetic analysis showed that the gene had a close evolutionary relationship with the avian herperviruses, and DEV should be placed into a single cluster within the subfamily Alphaherpesvirinae. Antigen prediction indicated that several potential B-cell epitopes sites located in the UL31 protein. To further study, the UL31 gene was cloned into a pET prokaryotic expression vector and transformed into Escherichia coli BL21 (DE3). A 55 kDa fusion protein was induced by the further culture at 37°C after addition of 0.8 mM IPTG. Polyclonal antibody raised against the recombinant UL31 from rabbit was prepared. A protein about 55 kDa that reacted with the antibody was detected in immunoblots of bacterial proteins, suggesting that the 55 kDa protein was the product of the UL31 gene. Immunofluorescence analysis revealed that the protein was localized in very fine punctate forms in the nuclei of infected cells. Our results may provide some insight for further research about the gene and also enrich the database of herpesvirus.

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Abbreviations

DEV:

Duck enteritis virus

HSV-1:

Herpes simplex virus 1

HCMV:

Human cytomegalovirus

EBV:

Epstein-barr virus

DVE:

Duck viral enteritis

HHV-3:

Human herpesvirus 3

MDV-1:

Marek’s disease virus type 1

CeHV-9:

Cercopithecine herpesvirus 9

MeHV-1:

Meleagrid herpesvirus 1

EHV-1:

Equid herpesvirus 1

BoHV-5:

Bovine herpesvirus 5

MDV-2:

Marek’s disease virus type 2

ILTV:

Infection laryngotracheitis virus

CeHV-16:

Cercopithecine herpesvirus 16

HHV-2:

Human herpesvirus 2

BoHV-1:

Bovine herpesvirus 1

CeHV-1:

Cercopithecine herpesvirus 1

SuHV-1:

Suid herpesvirus 1

HHV-1:

Human herpesvirus 1

HHV-7:

Human herpesvirus 7

PCMV:

Porcine cytomegalovirus

MCMV:

Murine cytomegalovirus

HHV-6:

Human herpesvirus 6

HHV-5:

Human herpesvirus 5

HHV-8:

Human herpesvirus 8

BoHV-4:

Bovine herpesvirus 4

SaHV-2:

Saimiriine herpesvirus 2

EHV-2:

Equid herpesvirus 2

MDPV:

Muscovy duck parvovirus

Prv:

Pseudorabies virus

Gpv:

Goose parvovirus

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Acknowledgments

The research was supported by the National Natural Science Foundation of China (30771598), Changjiang Scholars and Innovative Research Team in University(IRT0848), the earmarked fund for Modern Agro-industry Technology Research System (2009–2013), New Century Excellent Talents Program in University (NCET-06-0818), the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China (706050), the Cultivation Fund of the Key Scientific and Technical Innovation Project, department of Education of Sichuan Province (07ZZ028), Sichuan Province Outstanding Youths Fund (07ZQ026-132), and Sichuan Province Basic Research Program (07JY029-016/07JY029-017).

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Authors and Affiliations

  1. Avian Diseases Research Center, College of Veterinary Medicine, Sichuan Agricultural University, Ya’an, Sichuan, 625014, China

    Wei Xie & Anchun Cheng

  2. Key Laboratory of Animal Diseases and Human Health of Sichuan Province, Ya’an, Sichuan, 625014, China

    Anchun Cheng, Mingshu Wang, Hua Chang, Dekang Zhu & Qihui Luo

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  1. Wei Xie
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  2. Anchun Cheng
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  3. Mingshu Wang
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  4. Hua Chang
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Correspondence to Anchun Cheng.

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Xie, W., Cheng, A., Wang, M. et al. Molecular cloning and characterization of the UL31 gene from Duck enteritis virus. Mol Biol Rep 37, 1495–1503 (2010). https://doi.org/10.1007/s11033-009-9546-y

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