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Identification of a novel B-cell epitope specific for avian leukosis virus subgroup J gp85 protein

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Abstract

Avian leukosis virus subgroup J (ALV-J) is an avian oncogenic retrovirus that has caused severe economic losses in China. Gp85 protein is the main envelope protein and the most variable structural protein of ALV-J. It is also involved in virus neutralization. In this study, a specific monoclonal antibody, 4A3, was produced against the ALV-J gp85 protein. Immunofluorescence assays showed that 4A3 could react with different strains of ALV-J, including the British prototype isolate HPRS103, the American strains, an early Chinese broiler isolate, and layer isolates. A linear epitope on the gp85 protein was identified using a series of partially overlapping fragments spanning the gp85-encoding gene and subjecting them to western blot analysis. The results indicated that 134AEAELRDFI142 was the minimal linear epitope that could be recognized by mAb 4A3. Enzyme-linked immunosorbent assay (ELISA) revealed that chicken anti-ALV-J sera and mouse anti-ALV-J gp85 sera could also recognize the minimal linear epitope. Alignment analysis of amino acid sequences indicated that the epitope was highly conserved among 34 ALV-J strains. Furthermore, the epitope was not conserved among subgroup A and B of avian leukosis virus (ALV). Taken together, the mAb and the identified epitope may provide valuable tools for the development of new diagnostic methods for ALV-J.

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Acknowledgments

This study was funded by the Special Fund for Agro-Scientific Research in the Public Interest (no. 201203056) and the earmarked fund for the Modern Agro-industry Technology Research System (no. nycytx-42-G3-01).

Conflict of interest

The authors declare no conflict of interest.

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Authors and Affiliations

  1. Division of Avian Infectious Diseases, State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 427 Maduan Street, Nan Gang District, Harbin, 150001, Heilongjiang, People’s Republic of China

    Xiaofei Li, Qi Wang, Yanni Gao, Xiaole Qi, Yongqiang Wang, Honglei Gao, Yulong Gao & Xiaomei Wang

  2. Harbin National Engineering Research Center of Veterinary Biologics Co, Ltd, Harbin, 150001, China

    Haibo Zhu & Jiashan Sun

  3. College of Animal Medicine, Northeast Agriculture University, Harbin, 150001, China

    Jiashan Sun

  4. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Disease and Zoonoses, Yangzhou, 225009, China

    Xiaomei Wang

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  1. Xiaofei Li
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  2. Haibo Zhu
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  3. Qi Wang
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  4. Jiashan Sun
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  9. Yulong Gao
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Correspondence to Yulong Gao or Xiaomei Wang.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Materials 1 Primers used for expression of overlapping peptide fragments (PDF 194 kb)

Supplementary Materials 2 Viral strains used for IFA assay (PDF 122 kb)

705_2014_2318_MOESM3_ESM.pdf

Supplementary Materials 3 Viral strains used for amino acid sequence alignment analysis and their accession numbers in GenBank (PDF 138 kb)

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Li, X., Zhu, H., Wang, Q. et al. Identification of a novel B-cell epitope specific for avian leukosis virus subgroup J gp85 protein. Arch Virol 160, 995–1004 (2015). https://doi.org/10.1007/s00705-014-2318-6

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  • DOI: https://doi.org/10.1007/s00705-014-2318-6

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