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Immunoproteomic Analysis of Bordetella bronchiseptica Outer Membrane Proteins and Identification of New Immunogenic Proteins |
LIU Yan, QIN Feng-yan, BAO Guo-lian, CHEN Hui, XIAO Chen-wen, WEI Qiang , JI Quan-an |
Animal Husbandry and Veterinary Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, P.R.China |
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摘要 Bordetella bronchiseptica is a Gram-negative pathogen that causes acute and chronic respiratory infection in a variety of animals. To identify useful antigen candidates for diagnosis and subunit vaccine of B. bronchiseptica, immunoproteomic analysis was adopted to analyse outer membrane proteins of it. The outer membrane proteins extracted from B. bronchiseptica were separated by two-dimensional gel electrophoresis and analyzed by Western blotting for their reactivity with the convalescent serum against two strains. Immunogenic proteins were identified by matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS), a total of 14 proteins are common immunoreactive proteins, of which 1 was known antigen and 13 were novel immunogenic proteins for B. bronchiseptica. Putative lipoprotein gene was cloned and recombinantly expressed. The recombinant protein induced high titer antibody, but showed low protective indices against challenges with HB (B. bronchiseptica strain isolated from a infected rabbit). The mortality of mice was 80% compared to 100% of positive controls. The identification of these novel antigenic proteins is an important resource for further development of a new diagnostic test and vaccine for B. bronchiseptica.
Abstract Bordetella bronchiseptica is a Gram-negative pathogen that causes acute and chronic respiratory infection in a variety of animals. To identify useful antigen candidates for diagnosis and subunit vaccine of B. bronchiseptica, immunoproteomic analysis was adopted to analyse outer membrane proteins of it. The outer membrane proteins extracted from B. bronchiseptica were separated by two-dimensional gel electrophoresis and analyzed by Western blotting for their reactivity with the convalescent serum against two strains. Immunogenic proteins were identified by matrix-assisted laser desorption/ionization time of flight-mass spectrometry (MALDI-TOF-MS), a total of 14 proteins are common immunoreactive proteins, of which 1 was known antigen and 13 were novel immunogenic proteins for B. bronchiseptica. Putative lipoprotein gene was cloned and recombinantly expressed. The recombinant protein induced high titer antibody, but showed low protective indices against challenges with HB (B. bronchiseptica strain isolated from a infected rabbit). The mortality of mice was 80% compared to 100% of positive controls. The identification of these novel antigenic proteins is an important resource for further development of a new diagnostic test and vaccine for B. bronchiseptica.
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Received: 01 November 2012
Accepted:
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Fund: This work was supported by the China Agricultural Research System (nycytx-44-3-2), the Zhejiang Key Project on Agricultural Development through Science and Technology (2011C12028), the National Natural Science Foundation ofChina (31302068) and the Zhejiang Provincial Natural Science Foundation of China (LQ13C180002). |
Corresponding Authors:
BAO Guo-lian, Tel: +86-571-86404215, Fax: +86-571-86400836, E-mail: baoguolian@163.com
E-mail: baoguolian@163.com
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About author: LIU Yan, Mobile: 13136184186, E-mail: ly-liuyan@163.com |
Cite this article:
LIU Yan, QIN Feng-yan, BAO Guo-lian, CHEN Hui, XIAO Chen-wen, WEI Qiang , JI Quan-an.
2014.
Immunoproteomic Analysis of Bordetella bronchiseptica Outer Membrane Proteins and Identification of New Immunogenic Proteins. Journal of Integrative Agriculture, 13(9): 2010-2018.
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