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Transposon mutagenesis in Bordetella avium
R D Leyh1, R W Griffith, L H Arp
1Department of Veterinary Microbiology, Iowa State University, Ames 50011.
American Journal of Veterinary Research
|May 1, 1988
Summary
Transposon mutagenesis using Tn5 in Bordetella avium generated auxotrophic and hemagglutination-negative mutants. These stable mutations provide tools for studying B. avium genetics and virulence.
Area of Science:
- Microbiology
- Bacterial genetics
Background:
- Bordetella avium is a significant pathogen in poultry.
- Understanding B. avium genetics is crucial for developing control strategies.
Purpose of the Study:
- To develop a method for generating and identifying specific mutations in Bordetella avium.
- To characterize the nutritional requirements and hemagglutination properties of B. avium mutants.
Main Methods:
- Transposon mutagenesis using the Tn5 element was performed on B. avium.
- A defined minimal medium was utilized to screen for auxotrophic mutants.
- Conjugation from Escherichia coli SM10 via pSUP1011 facilitated Tn5 transfer.
Main Results:
- Approximately 1% of screened mutants exhibited auxotrophy, categorized into 13 distinct classes.
- Two classes of hemagglutination-negative mutants were successfully isolated.
- Tn5 insertions demonstrated high stability with reversion frequencies below 10(-7).
Conclusions:
- Transposon mutagenesis is an effective tool for generating stable mutations in B. avium.
- The identified auxotrophic and hemagglutination-negative mutants are valuable for further genetic studies.
- This work lays the foundation for future research into B. avium virulence factors and host interactions.