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Updated: May 22, 2026
Porphyromonas gingivalis as a Model Organism for Assessing Interaction of Anaerobic Bacteria with Host Cells
Published on: December 17, 2015
[Construction PG0839 gene-defective mutant of Porphyromonas gingivalis]
Jingbo Liu1, Yaping Pan, Chen Li
1Dept. of Periodontology, School of Stomatology, China Medical University, Shenyang 110002, China.
Objective:
In order to determine the function of PG0839 gene from Porphyromonas gingivalis (P. gingivalis) W83 strains, we intended to create a mutant in the PG0839 gene by homologous recombination.
Methods:
1 584bp PG0839 gene fragment was amplified, digested by BamH I and EcoR I, purified and ligated to pUC19. The recombinant plasmid was designated as pPG0839-1. The erm cassette (2 101 bp) was inserted into the EcoR V restriction site of the PG0839 gene. The resultant recombinant plasmid, pPG0839-2, was used as a donor in the electroporation of P. gingivalis W83. After electroporated and selected on erythromycin brain heart infusion plates, a single colony was collected and designated as PG0839 gene-defective mutant.
Results:
A mutant in PG0839 gene was created by insertional inactivation, and inactivation of PG0839 gene was confirmed by restriction endonuclease digestive, sequencing, polymerase chain reaction (PCR) and reverse transcription PCR.
Conclusion:
A PG0839 gene-defective mutant was created successfully.
Insights
Researchers successfully created a Porphyromonas gingivalis PG0839 gene-defective mutant using homologous recombination. This genetic modification is confirmed through various molecular techniques, paving the way for functional studies.
Area of Science:
- Microbiology
- Molecular Biology
- Genetics
Background:
- Porphyromonas gingivalis (P. gingivalis) is a key pathogen in periodontitis.
- Understanding the function of specific genes, like PG0839, is crucial for elucidating virulence mechanisms.
Purpose of the Study:
- To generate a PG0839 gene-defective mutant of P. gingivalis W83.
- To facilitate future studies on the role of the PG0839 gene in P. gingivalis.
Main Methods:
- Homologous recombination was employed to create the mutant.
- A PG0839 gene fragment was amplified, cloned into pUC19, and an erythromycin resistance cassette was inserted.
- Electroporation into P. gingivalis W83 followed by selection yielded the gene-defective mutant.
Main Results:
- A PG0839 gene-defective mutant was successfully generated through insertional inactivation.
- Confirmation of gene inactivation was achieved via restriction digestion, sequencing, PCR, and RT-PCR.
Conclusions:
- A PG0839 gene-defective mutant of P. gingivalis W83 was successfully created.
- The established mutant provides a valuable tool for investigating the PG0839 gene's function.
