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Transposition of Tn4351 in Porphyromonas gingivalis
C I Hoover1, E Abarbarchuk, C Y Ng
1Department of Stomatology, University of California, San Francisco 94143-0512.
Plasmid
|May 1, 1992
Summary
Transposon Tn4351 successfully integrated into the Porphyromonas gingivalis genome, enabling genetic manipulation. This advancement overcomes previous limitations in studying this bacterium, crucial for periodontitis research.
Area of Science:
- Microbiology
- Bacterial Genetics
- Molecular Biology
Background:
- Genetic analysis of Porphyromonas gingivalis is challenging due to a lack of natural genetic tools.
- Previous studies demonstrated Tn4351 transposition in Bacteroides uniformis using a suicide vector.
Purpose of the Study:
- To investigate the utility of transposon Tn4351 for genetic manipulation of Porphyromonas gingivalis.
- To establish a method for generating well-defined mutants of P. gingivalis.
Main Methods:
- Conjugation of Escherichia coli HB101 containing plasmid R751::*omega 4 with P. gingivalis 33277.
- Selection of erythromycin-resistant transconjugants.
- Southern blot hybridization to analyze the integration of Tn4351 and R751 sequences.
Main Results:
- Erythromycin-resistant P. gingivalis transconjugants were obtained at a frequency of 1.6 x 10(-7).
- Approximately 50% of transconjugants showed simple Tn4351 insertions, while the other 50% contained both Tn4351 and R751 sequences.
- Integration sites of Tn4351 varied across different transconjugants, indicating chromosomal insertion.
Conclusions:
- Transposon Tn4351 can transpose into the P. gingivalis chromosome.
- Tn4351 is a promising tool for generating insertion mutants in P. gingivalis.
- This method facilitates genetic studies of P. gingivalis, a key pathogen in periodontitis.