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Transposon Tn916 mutagenesis in Bacillus anthracis
B E Ivins1, S L Welkos, G B Knudson
1Division of Bacteriology, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland.
Infection and Immunity
|January 1, 1988
Summary
The streptococcal tetracycline resistance transposon Tn916 can be transferred to Bacillus anthracis, enabling genetic manipulation. This transposon facilitates mutagenesis, aiding in the study of B. anthracis genetics and virulence.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Bacillus anthracis is a significant human pathogen.
- Genetic manipulation of B. anthracis is crucial for understanding its virulence and developing countermeasures.
- Transposons offer a tool for introducing genetic modifications.
Purpose of the Study:
- To describe the mutagenesis of Bacillus anthracis using the streptococcal tetracycline resistance transposon Tn916.
- To establish efficient methods for transferring Tn916 into B. anthracis.
- To investigate the potential of Tn916 for generating auxotrophic mutants in B. anthracis.
Main Methods:
- Conjugation experiments were performed using filter mating procedures.
- Pretreatment of donor and recipient cells with nafcillin enhanced transfer frequency.
- Hybridization experiments confirmed the presence of the tetM gene in transconjugants.
Main Results:
- Tn916 was successfully transferred from Streptococcus faecalis to B. anthracis.
- Nafcillin pretreatment significantly increased the frequency of Tn916 transfer.
- Tn916 was also transferred to Bacillus subtilis.
- Mutagenesis of B. anthracis resulted in auxotrophic mutants, including phenylalanine auxotrophs.
Conclusions:
- Tn916 is an effective tool for mutagenesis in Bacillus anthracis.
- Optimized conjugation conditions enhance transposon transfer efficiency.
- Tn916-mediated mutagenesis can generate valuable auxotrophic mutants for genetic studies of B. anthracis.