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Journal of Bacteriology, October 2009, p. 5988-6002, Vol. 191, No. 19
0021-9193/09/$08.00+0     doi:10.1128/JB.00469-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Characterization and Comparative Analysis of the Genes Encoding Haemophilus parasuis Outer Membrane Proteins P2 and P5{triangledown} ,{dagger}

Michael A. Mullins,1 Karen B. Register,1* Darrell O. Bayles,2 Crystal L. Loving,1 Tracy L. Nicholson,1 Susan L. Brockmeier,1 David W. Dyer,3 and Gregory J. Phillips4

Respiratory Diseases of Livestock Research Unit,1 Bacterial Diseases of Livestock Research Unit, USDA Agricultural Research Service National Animal Disease Center, P.O. Box 70, Ames, Iowa 50010,2 Microbial Pathogenesis and Microbial Genomics, University of Oklahoma Health Sciences Center Biomedical Research Center, Oklahoma City, Oklahoma 73104,3 Department of Veterinary Microbiology and Preventive Medicine, Iowa State University College of Veterinary Medicine, Ames, Iowa 500114

Received 6 April 2009/ Accepted 17 July 2009

Haemophilus parasuis is a swine pathogen of significant industry concern, but little is known about how the organism causes disease. A related human pathogen, Haemophilus influenzae, has been better studied, and many of its virulence factors have been identified. Two of these, outer membrane proteins P2 and P5, are known to have important virulence properties. The goals of this study were to identify, analyze, and compare the genetic relatedness of orthologous genes encoding P2 and P5 proteins in a diverse group of 35 H. parasuis strains. Genes encoding P2 and P5 proteins were detected in all H. parasuis strains evaluated. The predicted amino acid sequences for both P2 and P5 proteins exhibit considerable heterogeneity, particularly in regions corresponding to predicted extracellular loops. Twenty-five variants of P2 and 17 variants of P5 were identified. The P2 proteins of seven strains were predicted to contain a highly conserved additional extracellular loop compared to the remaining strains and to H. influenzae P2. Antigenic-site predictions coincided with predicted extracellular loop regions of both P2 and P5. Neighbor-joining trees constructed using P2 and P5 sequences predicted divergent evolutionary histories distinct from those predicted by a multilocus sequence typing phylogeny based on partial sequencing of seven housekeeping genes. Real-time reverse transcription-PCR indicated that both genes are expressed in all of the strains.

* Corresponding author. Mailing address: Respiratory Diseases of Livestock Research Project, USDA/ARS/National Animal Disease Center, P.O. Box 70, 2300 Dayton Road, Ames, IA 50010. Phone: (515) 663-7700. Fax: (515) 663-7458. E-mail: Karen.Register{at}ars.usda.gov

{triangledown} Published ahead of print on 24 July 2009.

{dagger} Supplemental material for this article may be found at http://jb.asm.org/.

Journal of Bacteriology, October 2009, p. 5988-6002, Vol. 191, No. 19
0021-9193/09/$08.00+0     doi:10.1128/JB.00469-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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