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Multiplex PCR Assay for Typing of Staphylococcal Cassette Chromosome Mec Types I to V in Methicillin-resistant Staphylococcus aureus
Published on: September 5, 2013
Genetic analysis of Staphylococcus aureus with Tn4001.
G G Mahairas1, B R Lyon, R A Skurray
1Department of Microbiology, Iowa State University, Ames 50011.
Journal of Bacteriology
|July 1, 1989
Summary
The Tn4001 transposon facilitates gentamicin resistance in Staphylococcus aureus by moving to various DNA locations. Researchers developed efficient methods to isolate these insertions, aiding genetic studies.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Tn4001 is a composite transposon conferring antibiotic resistance in Staphylococcus aureus.
- Understanding transposon behavior is crucial for combating antibiotic resistance.
Purpose of the Study:
- To develop and optimize methods for isolating Tn4001 chromosomal insertions in S. aureus.
- To characterize the transposition and excision of Tn4001.
Main Methods:
- Isolation of Tn4001 insertions using UV irradiation and selection for thermoresistant, gentamicin-resistant (Gmr) isolates.
- Comparison of transposition efficiency in recombination-proficient and deficient hosts.
- Analysis of insertional auxotrophs and their reversion frequencies.
Main Results:
- Efficient isolation of Tn4001 transposition events, especially in recombination-deficient hosts.
- Successful generation of insertional auxotrophs with low reversion frequencies.
- Identification of precise Tn4001 excision and intergenic suppression as mechanisms for prototrophic reversion.
Conclusions:
- Tn4001 transposition can be efficiently isolated from S. aureus chromosomes.
- The developed methods facilitate genetic manipulation and the study of antibiotic resistance mechanisms.
- Tn4001 excision and suppression contribute to strain variability and adaptation.
