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A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits
Published on: February 16, 2014
Comparative functional genomic analysis of Pasteurellaceae adhesins using phage display
Lisa M Mullen1, Sean P Nair, John M Ward
1Division of Microbial Diseases, UCL Eastman Dental Institute, University College London, London WC1X 8LD, United Kingdom. l.mullen@eastman.ucl.ac.uk
Veterinary Microbiology
|January 30, 2007
Summary
Researchers identified novel bacterial adhesin genes in Pasteurellaceae pathogens using phage display. This discovery advances understanding of bacterial adhesion and virulence factors in animal disease.
Area of Science:
- Microbiology
- Bacterial Pathogenesis
- Molecular Biology
Background:
- The Pasteurellaceae family includes significant animal pathogens causing diverse diseases.
- Bacterial adhesion is a critical virulence factor, yet adhesins in Pasteurellaceae are poorly understood.
- Extracellular matrix components are common targets for bacterial adhesins.
Purpose of the Study:
- To identify genes encoding adhesins targeting host extracellular matrix molecules in key Pasteurellaceae species.
- To investigate the potential role of these identified genes as novel adhesins.
Main Methods:
- Construction of phage display libraries from genomic DNA of Haemophilus influenzae, Actinobacillus pleuropneumoniae, Pasteurella multocida, and Aggregatibacter actinomycetemcomitans.
- Screening of libraries against human/porcine fibronectin, serum albumin, and a commercial extracellular matrix (collagen, laminin, heparin sulfate).
- Identification and characterization of genes encoding putative adhesins.
Main Results:
- Four novel genes encoding putative adhesins were identified.
- These include a 34 kDa serum albumin-binding protein from H. influenzae and a 12.8 kDa fibronectin-binding protein from P. multocida.
- Additionally, a 13.7 kDa fibronectin-binding protein from A. actinomycetemcomitans and a 9.5 kDa serum albumin-binding protein from A. pleuropneumoniae were found.
Conclusions:
- Phage display is effective for identifying novel adhesin genes in Pasteurellaceae.
- The identified genes represent potential new adhesins previously not associated with this function.
- This research provides insights into bacterial adhesion mechanisms and potential virulence factors in animal pathogens.
