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Fifteen novel immunoreactive proteins of Chinese virulent Haemophilus parasuis serotype 5 verified by an immunoproteomic assay
Y. Yu, G. Wu, Z. Zhai, H. Yao, C. Lu, W. Zhang,
Language English Country United States
Document type Journal Article, Research Support, Non-U.S. Gov't
- MeSH
- Electrophoresis, Gel, Two-Dimensional MeSH
- Bacterial Proteins chemistry genetics immunology MeSH
- Haemophilus parasuis chemistry classification genetics immunology MeSH
- Haemophilus Infections immunology microbiology veterinary MeSH
- Mass Spectrometry MeSH
- Rabbits MeSH
- Molecular Sequence Data MeSH
- Swine Diseases immunology microbiology MeSH
- Swine MeSH
- Proteomics MeSH
- Antibodies, Bacterial analysis immunology MeSH
- Serogroup MeSH
- Blotting, Western MeSH
- Animals MeSH
- Check Tag
- Rabbits MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Geographicals
- China MeSH
Haemophilus parasuis (H. parasuis) is associated with meningitis, polyserositis, polyarthritis and bacterial pneumonia. At present, its prevention and control is difficult because of the lack of suitable subunit vaccines. Nowadays, high-throughput methods, immunoproteomics, are available to screen for more vaccine candidates. A protein extraction method for H. parasuis and two-dimensional electrophoresis (2-DE) were optimized to provide high-resolution profiles covering pH 3 to 10. Twenty immunoreactive spots were excised from gels after strict comparison between 2-DE Western blot membranes and the relevant gels. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) and MALDI-TOF-TOF-MS successfully identified 16 different proteins. Fifteen of them were reported as immunoreactive proteins in H. parasuis for the first time. In addition, recombinant HP5-7 (ABC transporter, periplasmic-binding protein) showed immunoreactivity both with hyperimmune rabbit serum and convalescent swine serum. Four recombinants of the 14 successfully expressed genes showed immunoreactivity with hyperimmune rabbit serum.
Key Lab of Animal Bacteriology Ministry of Agriculture Nanjing Agricultural University Nanjing China
References provided by Crossref.org
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