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ORIGINAL RESEARCH article

Front. Microbiol.
Sec. Microbiotechnology
Volume 15 - 2024 | doi: 10.3389/fmicb.2024.1476171
This article is part of the Research Topic Synthetic Biology for Non-Model Microbes View all articles

A Simple and Cost-effective Transformation System for Porphyromonas gingivalis via Natural Competence

Provisionally accepted
Kimihiro Abe Kimihiro Abe 1,2*Hiroko Yahara Hiroko Yahara 3*Ryoma Nakao Ryoma Nakao 2*Takehiro Yamaguchi Takehiro Yamaguchi 2Yukihiro Akeda Yukihiro Akeda 2
  • 1 National Institute of Infectious Diseases (NIID), Tokyo, Japan
  • 2 Department of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan
  • 3 Genome Medical Science Project, Research Institute, Tokyo, Japan

The final, formatted version of the article will be published soon.

    Porphyromonas gingivalis is a major oral bacterial pathogen responsible for severe periodontal diseases. Numerous studies have used genetic approaches to elucidate the molecular mechanisms underlying its pathogenicity. Typically, electroporation and conjugation are utilized for mutagenesis of P. gingivalis; however, these techniques require specialized equipment such as high-voltage electroporators, conjugative plasmids and donor strains. In this study, we present a simple, cost-effective transformation method for P. gingivalis without any special equipment by exploiting its natural DNA competence.P. gingivalis ATCC 33277 was grown to the early-exponential phase and mixed with a donor DNA cassette. This mixture was then spotted onto a BHI-HM blood-agar plate and incubated for one day to promote colony biofilm formation. The resulting colony biofilm was suspended in a liquid medium and spread onto antibiotic-containing agar plates.Transformants appeared within 4 to 5 days, achieving a maximum efficiency of 7.710 6 CFU/g. Although we optimized the transformation conditions using a representative strain ATCC 33277, but the method was also effective for other P. gingivalis strains, W83 and TDC60. Additionally, we discovered that deletion of PGN_0421 or PGN_0519, encoding putative ComEA and ComEC, abolished competency, indicating that these gene products are essential for the natural competence.

    Keywords: P. gingivalis Transformation Porphyromonas gingivalis, Natural competence, transformation, horizontal gene transfer, Genetic Engineering

    Received: 05 Aug 2024; Accepted: 03 Oct 2024.

    Copyright: © 2024 Abe, Yahara, Nakao, Yamaguchi and Akeda. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Kimihiro Abe, National Institute of Infectious Diseases (NIID), Tokyo, Japan
    Hiroko Yahara, Genome Medical Science Project, Research Institute, Tokyo, Japan
    Ryoma Nakao, Department of Bacteriology I, National Institute of Infectious Diseases, Tokyo, Japan

    Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.