Purification of Mfa1 and FimA Fimbriae from P. gingivalis Mutants

Mfa1 and FimA fimbriae were purified from P. gingivalis mutant JI-1 (fimA deleted) [36 (link),37 (link)] and SMF-1 (mfa1 deleted) [38 (link),39 (link)] derived from ATCC 33277. JI-1 and SMF-1 cells were cultured under anaerobic conditions at 37 °C on Brucella HK agar medium (KYOKUTO, Tokyo, Japan) prepared by mixing 5% [volume/volume (v/v)] laked rabbit blood, 2.5 μg/mL hemin (Sigma-Aldrich, St. Louis, MO, USA), 5 μg/mL menadione (Sigma-Aldrich), and distilled water. Liquid medium was prepared by mixing trypticase soy broth (Thermo Fisher Scientific), 0.25% yeast extract (Thermo Fisher Scientific), and distilled water, followed by sterilization and the addition of 2.5 μg/mL hemin and 5 μg/mL menadione.

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Suzuki Y., Kikuchi T., Goto H., Takayanagi Y., Kawamura S., Sawada N., Naiki Y., Kondo H., Hayashi J.I., Hasegawa Y, & Mitani A. (2022). Porphyromonas gingivalis Fimbriae Induce Osteoclastogenesis via Toll-like Receptors in RAW264 Cells. International Journal of Molecular Sciences, 23(23), 15293.

Publication 2022

hemin menadione trypticase soy broth yeast extract

Agar Blood Brucella Cells Fimbriae Hemin Menadione Rabbit Sterilization Trypticase soy broth Yeast

Corresponding Organization : Aichi Gakuin University

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Variable analysis

independent variables

Deletion of fimA gene in P. gingivalis mutant JI-1
Deletion of mfa1 gene in P. gingivalis mutant SMF-1

dependent variables

Purification of Mfa1 and FimA fimbriae from P. gingivalis mutants JI-1 and SMF-1, respectively

control variables

Culture conditions for JI-1 and SMF-1 cells (anaerobic conditions at 37 °C on Brucella HK agar medium with 5% laked rabbit blood, 2.5 μg/mL hemin, and 5 μg/mL menadione)
Liquid medium composition (trypticase soy broth, 0.25% yeast extract, 2.5 μg/mL hemin, and 5 μg/mL menadione)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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