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  • Title: Purification of Native Mfa1 Fimbriae from Porphyromonas gingivalis.
    Author: Hasegawa Y, Nagano K, Murakami Y, Lamont RJ.
    Journal: Methods Mol Biol; 2021; 2210():75-86. PubMed ID: 32815129.
    Abstract:
    Fimbriae of the periodontal pathogen Porphyromonas gingivalis mediate its colonization through associations with other bacteria and host tissues. P. gingivalis generally expresses two distinct fimbrial types, FimA and Mfa1. In P. gingivalis ATCC 33277, FimA fimbriae are present as long filaments easily detached from cells, whereas Mfa1 fimbriae are short filaments compactly bound to the cell surface. Because of this unique characteristic, FimA fimbriae have been selectively and easily isolated from the bacterial cell surface through mechanical shearing such as by pipetting and stirring. However, P. gingivalis ATCC 33277 harbors a mutation in the gene encode the fimbrial length regulator, FimB, and thus produces unusually long FimA fimbriae length. Hence, mechanical shearing to remove FimA is potentially applicable only for this type strain. Here we present protocols to purify intact Mfa1 fimbriae from a fimA-deficient mutant strain. Mfa1 fimbriae are purified from cell lysates, using a French pressure cell and through ion-exchange chromatography. The purity of Mfa1 fimbriae can be confirmed through sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblotting, and electron microscopy.
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