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    ATCC p gingivalis atcc 33277
    Macrophage migration-inhibitory factor (MIF) acted as a regulator in Porphyromonas <t>gingivalis</t> ( P. gingivalis )-induced intercellular adhesion molecule-1 (ICAM-1) expression. a Western blot analysis of ICAM-1 in EA.hy926 cells. b Quantitative analyses of the optical density relative to the internal reference protein GAPDH in Western blot analysis. c Quantitative Real-time PCR analysis of ICAM-1 mRNA. Infection with P. gingivalis <t>ATCC</t> 33277 (MOI = 100, 24 h) could induce the expression of ICAM-1 and ICAM-1 mRNA in EA.hy926 cells, this inductive effect of P. gingivalis was blocked by the MIF antagonist ISO-1. Sufficient exogenous recombinant MIF (rMIF) supplementation rescued the ICAM-1 expression. Data are presented as means ± SD from three independent experiments. * P
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    Macrophage migration-inhibitory factor (MIF) acted as a regulator in Porphyromonas gingivalis ( P. gingivalis )-induced intercellular adhesion molecule-1 (ICAM-1) expression. a Western blot analysis of ICAM-1 in EA.hy926 cells. b Quantitative analyses of the optical density relative to the internal reference protein GAPDH in Western blot analysis. c Quantitative Real-time PCR analysis of ICAM-1 mRNA. Infection with P. gingivalis ATCC 33277 (MOI = 100, 24 h) could induce the expression of ICAM-1 and ICAM-1 mRNA in EA.hy926 cells, this inductive effect of P. gingivalis was blocked by the MIF antagonist ISO-1. Sufficient exogenous recombinant MIF (rMIF) supplementation rescued the ICAM-1 expression. Data are presented as means ± SD from three independent experiments. * P

    Journal: BMC Microbiology

    Article Title: Porphyromonas gingivalis ATCC 33277 promotes intercellular adhesion molecule-1 expression in endothelial cells and monocyte-endothelial cell adhesion through macrophage migration inhibitory factor

    doi: 10.1186/s12866-018-1156-1

    Figure Lengend Snippet: Macrophage migration-inhibitory factor (MIF) acted as a regulator in Porphyromonas gingivalis ( P. gingivalis )-induced intercellular adhesion molecule-1 (ICAM-1) expression. a Western blot analysis of ICAM-1 in EA.hy926 cells. b Quantitative analyses of the optical density relative to the internal reference protein GAPDH in Western blot analysis. c Quantitative Real-time PCR analysis of ICAM-1 mRNA. Infection with P. gingivalis ATCC 33277 (MOI = 100, 24 h) could induce the expression of ICAM-1 and ICAM-1 mRNA in EA.hy926 cells, this inductive effect of P. gingivalis was blocked by the MIF antagonist ISO-1. Sufficient exogenous recombinant MIF (rMIF) supplementation rescued the ICAM-1 expression. Data are presented as means ± SD from three independent experiments. * P

    Article Snippet: In contrast, cell adhesion was decreased in ISO-1-treated cells compared with those infected with P. gingivalis ATCC 33277 ( P < 0.01).

    Techniques: Migration, Expressing, Western Blot, Real-time Polymerase Chain Reaction, Infection, Recombinant

    Macrophage migration-inhibitory factor (MIF) regulated the increased monocyte adhesion to endothelial cells infected with Porphyromonas gingivalis ( P. gingivalis ) . a THP-1 cell adhesion to EA.hy926 cells was labeled by calcein-AM and visualized. Pictures are representative fields captured by flurencence microscope (upper line) or microscope (lower line) of three independent experiments (magnification × 100). b The histogram of the evaluation of adhered THP-1 cell assessed by cell count assay. Compared with uninfected cells, P. gingivalis ATCC 33277 infection (MOI = 100, 24 h) markedly increased THP-1 cell adhesion to endothelial cells ( P

    Journal: BMC Microbiology

    Article Title: Porphyromonas gingivalis ATCC 33277 promotes intercellular adhesion molecule-1 expression in endothelial cells and monocyte-endothelial cell adhesion through macrophage migration inhibitory factor

    doi: 10.1186/s12866-018-1156-1

    Figure Lengend Snippet: Macrophage migration-inhibitory factor (MIF) regulated the increased monocyte adhesion to endothelial cells infected with Porphyromonas gingivalis ( P. gingivalis ) . a THP-1 cell adhesion to EA.hy926 cells was labeled by calcein-AM and visualized. Pictures are representative fields captured by flurencence microscope (upper line) or microscope (lower line) of three independent experiments (magnification × 100). b The histogram of the evaluation of adhered THP-1 cell assessed by cell count assay. Compared with uninfected cells, P. gingivalis ATCC 33277 infection (MOI = 100, 24 h) markedly increased THP-1 cell adhesion to endothelial cells ( P

    Article Snippet: In contrast, cell adhesion was decreased in ISO-1-treated cells compared with those infected with P. gingivalis ATCC 33277 ( P < 0.01).

    Techniques: Migration, Infection, Labeling, Microscopy, Cell Counting

    Porphyromonas gingivalis ( P. gingivalis ) ATCC 33277 infection enhances MIF secretion in EA.hy926 cells . EA.hy926 were challenged with Escherichia coli ( E. coli ) lipopolysaccharide (LPS, 1 μg/mL) or P. gingivalis (MOI = 100) for 4, 10 or 24 h. MIF levels was analyzed using ELISA. Infection of P. gingivalis significantly increased MIF secretion in EA.hy926 cells. EA.hy926 were incubated with medium only as a control. E.coli LPS was used as a positive control. The I bar shows the standard deviation. * P

    Journal: BMC Microbiology

    Article Title: Porphyromonas gingivalis ATCC 33277 promotes intercellular adhesion molecule-1 expression in endothelial cells and monocyte-endothelial cell adhesion through macrophage migration inhibitory factor

    doi: 10.1186/s12866-018-1156-1

    Figure Lengend Snippet: Porphyromonas gingivalis ( P. gingivalis ) ATCC 33277 infection enhances MIF secretion in EA.hy926 cells . EA.hy926 were challenged with Escherichia coli ( E. coli ) lipopolysaccharide (LPS, 1 μg/mL) or P. gingivalis (MOI = 100) for 4, 10 or 24 h. MIF levels was analyzed using ELISA. Infection of P. gingivalis significantly increased MIF secretion in EA.hy926 cells. EA.hy926 were incubated with medium only as a control. E.coli LPS was used as a positive control. The I bar shows the standard deviation. * P

    Article Snippet: In contrast, cell adhesion was decreased in ISO-1-treated cells compared with those infected with P. gingivalis ATCC 33277 ( P < 0.01).

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Incubation, Positive Control, Standard Deviation

    rprY and pat are co-transcribed as an operon in P. gingivalis ATCC 33277. (a) Schematic of the rprY-pat locus in strain ATCC 33277. Locations of RT-PCR primers are indicated by small arrows and corresponding amplicons are numbered. (B and C) RT-PCR assays for expression of rprY-pat genes. (b) G enomic DNA was used as the positive control template for PCR. Amplicon numbers correspond to those indicated in panel A. (c) cDNA was used as the template for PCR.

    Journal: Journal of Oral Microbiology

    Article Title: Post-translational regulation of a Porphyromonas gingivalis regulator

    doi: 10.1080/20002297.2018.1487743

    Figure Lengend Snippet: rprY and pat are co-transcribed as an operon in P. gingivalis ATCC 33277. (a) Schematic of the rprY-pat locus in strain ATCC 33277. Locations of RT-PCR primers are indicated by small arrows and corresponding amplicons are numbered. (B and C) RT-PCR assays for expression of rprY-pat genes. (b) G enomic DNA was used as the positive control template for PCR. Amplicon numbers correspond to those indicated in panel A. (c) cDNA was used as the template for PCR.

    Article Snippet: In a previous report, we presented data indicating that the response regulator RprY of P. gingivalis strain ATCC 33277 played a role as a repressor in the oxidative stress response, with genes encoding AhpC, GroES, ClpB, and DnaK included in its regulon [ ].

    Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Positive Control, Polymerase Chain Reaction, Amplification

    The arrangement of porR and its neighbouring genes in P. gingivalis ATCC 33277 genome. porR (PGN_1236) and its neighbouring genes are flanked by IS5 family transposons that formed a composite transposon of 70 kbp in length. The genes that involve in biosynthesis of A-LPS are represented by yellow rectangles while the gene that does not involve is represented by brown rectangle. The genes for hypothetical proteins are represented by white rectangles. The genes for IS5 family transposases are represented by cyan rectangles. The purple triangles represented 12 bp inverted repeats that flanked the genes for IS5 family transposases. Name of proteins encoded by the genes are shown under rectangles that represented the genes. The slashes indicated gaps in the genome.

    Journal: PeerJ

    Article Title: Phylogenetic comparison between Type IX Secretion System (T9SS) protein components suggests evidence of horizontal gene transfer

    doi: 10.7717/peerj.9019

    Figure Lengend Snippet: The arrangement of porR and its neighbouring genes in P. gingivalis ATCC 33277 genome. porR (PGN_1236) and its neighbouring genes are flanked by IS5 family transposons that formed a composite transposon of 70 kbp in length. The genes that involve in biosynthesis of A-LPS are represented by yellow rectangles while the gene that does not involve is represented by brown rectangle. The genes for hypothetical proteins are represented by white rectangles. The genes for IS5 family transposases are represented by cyan rectangles. The purple triangles represented 12 bp inverted repeats that flanked the genes for IS5 family transposases. Name of proteins encoded by the genes are shown under rectangles that represented the genes. The slashes indicated gaps in the genome.

    Article Snippet: Arrangement of porR and its neighbouring genes in P. gingivalis ATCC 33277 genome As shown in , porR and its neighbouring genes are flanked by IS5 family transposons.

    Techniques:

    Biofilm-forming capacity of P. gingivalis ATCC 33277 is reduced compared with that of vimA mutant FLL451. FLL451 showed a five times higher biofilm-forming capacity (∗, P

    Journal: Infection and Immunity

    Article Title: VimA-Dependent Modulation of Acetyl Coenzyme A Levels and Lipid A Biosynthesis Can Alter Virulence in Porphyromonas gingivalis

    doi: 10.1128/IAI.06062-11

    Figure Lengend Snippet: Biofilm-forming capacity of P. gingivalis ATCC 33277 is reduced compared with that of vimA mutant FLL451. FLL451 showed a five times higher biofilm-forming capacity (∗, P

    Article Snippet: DNA fragments containing an upstream regulatory region and open reading frame (ORF) for vimA were amplified from P. gingivalis ATCC 33277 chromosomal DNA using the appropriate primer set (Table S1 in the supplemental material).

    Techniques: Mutagenesis

    (A) Polyacrylamide gel electrophoresis of the lipid A-associated proteins of P. gingivalis strains. Lane 1, P. gingivalis W83; lane 2, FLL92 ( P. gingivalis vimA mutant in W83); lane 3, FLL451 ( P. gingivalis vimA mutant in ATCC 33277; protein bands found

    Journal: Infection and Immunity

    Article Title: VimA-Dependent Modulation of Acetyl Coenzyme A Levels and Lipid A Biosynthesis Can Alter Virulence in Porphyromonas gingivalis

    doi: 10.1128/IAI.06062-11

    Figure Lengend Snippet: (A) Polyacrylamide gel electrophoresis of the lipid A-associated proteins of P. gingivalis strains. Lane 1, P. gingivalis W83; lane 2, FLL92 ( P. gingivalis vimA mutant in W83); lane 3, FLL451 ( P. gingivalis vimA mutant in ATCC 33277; protein bands found

    Article Snippet: DNA fragments containing an upstream regulatory region and open reading frame (ORF) for vimA were amplified from P. gingivalis ATCC 33277 chromosomal DNA using the appropriate primer set (Table S1 in the supplemental material).

    Techniques: Polyacrylamide Gel Electrophoresis, Mutagenesis

    Ultrastructural studies on the invasion of P. gingivalis strains. (A) Invasion of epithelial cells by P. gingivalis ATCC 33277 showing invasion into the cells; (B) invasion of P. gingivalis W83 showing intact morphology of the pathogen; (C) P. gingivalis

    Journal: Infection and Immunity

    Article Title: VimA-Dependent Modulation of Acetyl Coenzyme A Levels and Lipid A Biosynthesis Can Alter Virulence in Porphyromonas gingivalis

    doi: 10.1128/IAI.06062-11

    Figure Lengend Snippet: Ultrastructural studies on the invasion of P. gingivalis strains. (A) Invasion of epithelial cells by P. gingivalis ATCC 33277 showing invasion into the cells; (B) invasion of P. gingivalis W83 showing intact morphology of the pathogen; (C) P. gingivalis

    Article Snippet: DNA fragments containing an upstream regulatory region and open reading frame (ORF) for vimA were amplified from P. gingivalis ATCC 33277 chromosomal DNA using the appropriate primer set (Table S1 in the supplemental material).

    Techniques:

    Immunoblot analysis of the PGN_1234 mutant. Whole-cell lysates of wild-type P. gingivalis ATCC 33277, the PGN_1234 mutant, and the complemented PGN_1234 + strains were separated by SDS-PAGE and immunoblotted using various primary antibodies, as shown. MAb 1B5 detects A-LPS while MAb TDC-5-2-1 detects both A-LPS and O-LPS.

    Journal: mBio

    Article Title: Type IX Secretion System Cargo Proteins Are Glycosylated at the C Terminus with a Novel Linking Sugar of the Wbp/Vim Pathway

    doi: 10.1128/mBio.01497-20

    Figure Lengend Snippet: Immunoblot analysis of the PGN_1234 mutant. Whole-cell lysates of wild-type P. gingivalis ATCC 33277, the PGN_1234 mutant, and the complemented PGN_1234 + strains were separated by SDS-PAGE and immunoblotted using various primary antibodies, as shown. MAb 1B5 detects A-LPS while MAb TDC-5-2-1 detects both A-LPS and O-LPS.

    Article Snippet: The C-terminal coding region of the mfa2 gene (mfa2 C ) was amplified by PCR using the primer pair mfa2-F and mfa2-R and P. gingivalis ATCC 33277 genomic DNA as the template.

    Techniques: Mutagenesis, SDS Page

    Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with heat inactivated and viable P. gingivalis wild type strain ATCC 33277, ΔKgp, and ΔRgpArgpB mutants (a) and heat-inactivated and viable P. gingivalis wild-type strain W50, ΔKgp, and ΔRgpArgpB mutants and with addition of KYT-36 and KYT-1 peptides (b). *=significantly different from wild-type strain of the same MOI ( p

    Journal: Journal of Oral Microbiology

    Article Title: The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro

    doi: 10.3402/jom.v7.27543

    Figure Lengend Snippet: Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with heat inactivated and viable P. gingivalis wild type strain ATCC 33277, ΔKgp, and ΔRgpArgpB mutants (a) and heat-inactivated and viable P. gingivalis wild-type strain W50, ΔKgp, and ΔRgpArgpB mutants and with addition of KYT-36 and KYT-1 peptides (b). *=significantly different from wild-type strain of the same MOI ( p

    Article Snippet: Viable W83 and W50 inhibited cell migration more than their heat-inactivated variants (not statistically significant for W50 at MOI 10); however, no differences were found between heat-inactivated and viable P. gingivalis ATCC 33277 (p > 0.05).

    Techniques:

    Percentage closure of a scratch in oral epithelial cells challenged with P. gingivalis ATCC 33277 and medium only.

    Journal: Journal of Oral Microbiology

    Article Title: The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro

    doi: 10.3402/jom.v7.27543

    Figure Lengend Snippet: Percentage closure of a scratch in oral epithelial cells challenged with P. gingivalis ATCC 33277 and medium only.

    Article Snippet: Viable W83 and W50 inhibited cell migration more than their heat-inactivated variants (not statistically significant for W50 at MOI 10); however, no differences were found between heat-inactivated and viable P. gingivalis ATCC 33277 (p > 0.05).

    Techniques:

    Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with heat-inactivated and viable P. gingivalis strains ATCC 33277, W83, and W50. a=significantly different from control ( p

    Journal: Journal of Oral Microbiology

    Article Title: The impact of virulence factors of Porphyromonas gingivalis on wound healing in vitro

    doi: 10.3402/jom.v7.27543

    Figure Lengend Snippet: Mean relative closure (±SEM) from all biological replicates of scratches in oral epithelial cells challenged with heat-inactivated and viable P. gingivalis strains ATCC 33277, W83, and W50. a=significantly different from control ( p

    Article Snippet: Viable W83 and W50 inhibited cell migration more than their heat-inactivated variants (not statistically significant for W50 at MOI 10); however, no differences were found between heat-inactivated and viable P. gingivalis ATCC 33277 (p > 0.05).

    Techniques:

    Bacterial adhesion and invasion of OK ‐F6 monolayers by wild‐type, ∆ ompA1, ∆ ompA2 and ∆ ompA1A2 mutants. P. gingivalis was incubated with a monolayer of OK ‐F6 at a MOI 1:100 as described for invasion assays. Invasion was defined as the percentage of the inoculum protected from metronidazole killing. Total association was defined as the number of bacteria that have adhered to the OK ‐F6 cell and invaded. Adherence was calculated from subtracting invasion CFU s from the total association. Each % value was determined by calculating the CFU s recovered as a percentage of the viability of that strain, and corrected to wild‐type P. gingivalis total association (=1). Wild‐type and mutant strains were evaluated for invasion and adherence efficiency (A), and the complemented ompA2 mutant (B) assessed. Statistical significance was determined by students’ t ‐test and designated as * p

    Journal: MicrobiologyOpen

    Article Title: Role of OmpA2 surface regions of Porphyromonas gingivalis in host–pathogen interactions with oral epithelial cellsRole of OmpA2 surface regions of Porphyromonas gingivalis in host–pathogen interactions with oral epithelial cells

    doi: 10.1002/mbo3.401

    Figure Lengend Snippet: Bacterial adhesion and invasion of OK ‐F6 monolayers by wild‐type, ∆ ompA1, ∆ ompA2 and ∆ ompA1A2 mutants. P. gingivalis was incubated with a monolayer of OK ‐F6 at a MOI 1:100 as described for invasion assays. Invasion was defined as the percentage of the inoculum protected from metronidazole killing. Total association was defined as the number of bacteria that have adhered to the OK ‐F6 cell and invaded. Adherence was calculated from subtracting invasion CFU s from the total association. Each % value was determined by calculating the CFU s recovered as a percentage of the viability of that strain, and corrected to wild‐type P. gingivalis total association (=1). Wild‐type and mutant strains were evaluated for invasion and adherence efficiency (A), and the complemented ompA2 mutant (B) assessed. Statistical significance was determined by students’ t ‐test and designated as * p

    Article Snippet: We then used these peptides alongside wild‐type P. gingivalis ATCC 33277 in adhesion and invasion blocking studies to establish which OmpA2 loops are important in mediating interactions with host cells.

    Techniques: Incubation, Mutagenesis

    OmpA2 extracellular loops display direct binding to oral epithelial cells. Antibiotic protection assays were carried out with wild‐type P. gingivalis in the presence of each extracellular loop individually at 50 μg ml −1 (A), or at 50 μg ml −1 total concentration for all four loops (B). (C) Extracellular loop peptides were bound to NeutrAvidin ® ‐green fluorescent microspheres at 50 μg ml −1 and incubated with a monolayer of OK ‐F6 cells and the total fluorescence at 488 nm /515 nm (ex/em) recorded as a measure of the quantity of extracellular loop peptides bound to cells, relative to BSA ‐coated microspheres. (D) A scrambled peptide was used as a control. (E) Immunofluorescence images of peptide 4‐bound microspheres (P4) incubated with OK ‐F6 monolayers and imaged at ×100 magnification, BSA ‐coated microspheres ( BSA ) and scrambled‐peptide‐bound microspheres (P4‐S). NeutrAvidin ® ‐green microspheres are visualised in the Green channel (488 nm) with WGA ‐Texas Red ® (red, 549 nm) highlighting cell membranes and DAPI (blue) for cell nuclei. Statistical significance was determined by students’ t ‐test and designated as ** p

    Journal: MicrobiologyOpen

    Article Title: Role of OmpA2 surface regions of Porphyromonas gingivalis in host–pathogen interactions with oral epithelial cellsRole of OmpA2 surface regions of Porphyromonas gingivalis in host–pathogen interactions with oral epithelial cells

    doi: 10.1002/mbo3.401

    Figure Lengend Snippet: OmpA2 extracellular loops display direct binding to oral epithelial cells. Antibiotic protection assays were carried out with wild‐type P. gingivalis in the presence of each extracellular loop individually at 50 μg ml −1 (A), or at 50 μg ml −1 total concentration for all four loops (B). (C) Extracellular loop peptides were bound to NeutrAvidin ® ‐green fluorescent microspheres at 50 μg ml −1 and incubated with a monolayer of OK ‐F6 cells and the total fluorescence at 488 nm /515 nm (ex/em) recorded as a measure of the quantity of extracellular loop peptides bound to cells, relative to BSA ‐coated microspheres. (D) A scrambled peptide was used as a control. (E) Immunofluorescence images of peptide 4‐bound microspheres (P4) incubated with OK ‐F6 monolayers and imaged at ×100 magnification, BSA ‐coated microspheres ( BSA ) and scrambled‐peptide‐bound microspheres (P4‐S). NeutrAvidin ® ‐green microspheres are visualised in the Green channel (488 nm) with WGA ‐Texas Red ® (red, 549 nm) highlighting cell membranes and DAPI (blue) for cell nuclei. Statistical significance was determined by students’ t ‐test and designated as ** p

    Article Snippet: We then used these peptides alongside wild‐type P. gingivalis ATCC 33277 in adhesion and invasion blocking studies to establish which OmpA2 loops are important in mediating interactions with host cells.

    Techniques: Binding Assay, Concentration Assay, Incubation, Fluorescence, Immunofluorescence, Whole Genome Amplification

    Involvement of SigP in the transcription of T9SS components. ( A ) Correlation in gene expression between porX mutant (KDP363) and sigP mutant (KDP314). Gene expression was measured by the custom tiling microarrays spanning the whole genome of P. gingivalis ATCC 33277. The expression level for each coding sequence was normalized with the constant from the 16S rRNA gene and represented as a ratio to that from wild type. Experiments were performed three times with independently prepared labelled cDNAs. The genes encoding T9SS components are represented in red. ( B ) EMSA assay of the promoter regions of T9SS component genes by rSigP. Probes corresponding to the possible promoter regions were generated by PCR and labelled with digoxigenin. Binding specificity was tested by competition with 100-fold excess of the appropriate unlabelled probe. PGN numbers for genes are indicated in parenthesis.

    Journal: Scientific Reports

    Article Title: A two-component system regulates gene expression of the type IX secretion component proteins via an ECF sigma factor

    doi: 10.1038/srep23288

    Figure Lengend Snippet: Involvement of SigP in the transcription of T9SS components. ( A ) Correlation in gene expression between porX mutant (KDP363) and sigP mutant (KDP314). Gene expression was measured by the custom tiling microarrays spanning the whole genome of P. gingivalis ATCC 33277. The expression level for each coding sequence was normalized with the constant from the 16S rRNA gene and represented as a ratio to that from wild type. Experiments were performed three times with independently prepared labelled cDNAs. The genes encoding T9SS components are represented in red. ( B ) EMSA assay of the promoter regions of T9SS component genes by rSigP. Probes corresponding to the possible promoter regions were generated by PCR and labelled with digoxigenin. Binding specificity was tested by competition with 100-fold excess of the appropriate unlabelled probe. PGN numbers for genes are indicated in parenthesis.

    Article Snippet: Probes used in this assay were designed from the possible transcription starting sites in the tiling microarray, and PCR-amplified from P. gingivalis ATCC 33277 DNA using primers shown in .

    Techniques: Expressing, Mutagenesis, Sequencing, Generated, Polymerase Chain Reaction, Binding Assay

    Structure and localization of PorX and PorY. ( A ) Schematic representation of the functional domains of PorX and PorY. The symbols denote the following, RR, response regulatory domain; PglZ, PglZ motif; ALP, alkaline phosphatase-like core domain; SS, signal sequence; TM, transmembrane domain; HK1, histidine kinase phosphoacceptor domain; HK2, histidine kinase ATPase domain. Arrows below the schema indicate the recombinant proteins generated in this study. The domains were predicted by the Kyoto Encyclopedia of Genes and Genomes (KEGG) Sequence Similarity DataBase (KEGG SSDB). ( B ) Subcellular localization of PorX and PorY in P. gingivalis. Fractionated cell lysates of the wild-type strain ATCC 33277 were subjected to immunodetection with antisera against PorX and PorY. WC, whole cell lysate; C/P, cytoplasm/periplasm; CE, cell envelope; IM, inner membrane; OM, outer membrane. Arrowheads indicate the immunoreacting PorX and PorY bands. ( C ) SDS-PAGE profile of the purified recombinant proteins.

    Journal: Scientific Reports

    Article Title: A two-component system regulates gene expression of the type IX secretion component proteins via an ECF sigma factor

    doi: 10.1038/srep23288

    Figure Lengend Snippet: Structure and localization of PorX and PorY. ( A ) Schematic representation of the functional domains of PorX and PorY. The symbols denote the following, RR, response regulatory domain; PglZ, PglZ motif; ALP, alkaline phosphatase-like core domain; SS, signal sequence; TM, transmembrane domain; HK1, histidine kinase phosphoacceptor domain; HK2, histidine kinase ATPase domain. Arrows below the schema indicate the recombinant proteins generated in this study. The domains were predicted by the Kyoto Encyclopedia of Genes and Genomes (KEGG) Sequence Similarity DataBase (KEGG SSDB). ( B ) Subcellular localization of PorX and PorY in P. gingivalis. Fractionated cell lysates of the wild-type strain ATCC 33277 were subjected to immunodetection with antisera against PorX and PorY. WC, whole cell lysate; C/P, cytoplasm/periplasm; CE, cell envelope; IM, inner membrane; OM, outer membrane. Arrowheads indicate the immunoreacting PorX and PorY bands. ( C ) SDS-PAGE profile of the purified recombinant proteins.

    Article Snippet: Probes used in this assay were designed from the possible transcription starting sites in the tiling microarray, and PCR-amplified from P. gingivalis ATCC 33277 DNA using primers shown in .

    Techniques: Functional Assay, ALP Assay, Sequencing, Recombinant, Generated, Immunodetection, SDS Page, Purification

    Electron micrograph showing changes in surface structures of P. gingivalis ATCC 33277 and W83. Bacterial cells grown to the log phase (OD 600 of 0.7–0.9) were processed for electron microscopic examination using formvar-carbon coated grids (500 mesh) and were examined using Philips Tecnai 12 TEM. Fimbriae were lacking in the vimF mutant FLL476 when compared with the wild ATCC33277 and the complemented strain FLL476C’. A thick glycocalyx along with vesicles and a well-defined outer membrane was observed in W83. FLL95 showed hazy outer membrane with reduced visicles. In the complemented strain FLL95C’ the outer membrane morphology was restored.

    Journal: PLoS ONE

    Article Title: In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose

    doi: 10.1371/journal.pone.0063367

    Figure Lengend Snippet: Electron micrograph showing changes in surface structures of P. gingivalis ATCC 33277 and W83. Bacterial cells grown to the log phase (OD 600 of 0.7–0.9) were processed for electron microscopic examination using formvar-carbon coated grids (500 mesh) and were examined using Philips Tecnai 12 TEM. Fimbriae were lacking in the vimF mutant FLL476 when compared with the wild ATCC33277 and the complemented strain FLL476C’. A thick glycocalyx along with vesicles and a well-defined outer membrane was observed in W83. FLL95 showed hazy outer membrane with reduced visicles. In the complemented strain FLL95C’ the outer membrane morphology was restored.

    Article Snippet: The vimF -defective mutant (FLL476) in the P. gingivalis ATCC 33277 genetic background showed a phenotype similar to that of the vimF -defective mutant (FLL95) in the P. gingivalis W83 genetic background.

    Techniques: Transmission Electron Microscopy, Mutagenesis

    Comparison of biofilm formation, autoaggregation, hemagglutination and invasion assay. A. Biofilm formation of ATCC 33277, FLL476 and FLL476C’ were compared. Biofilm assay was done by staining adherent cells of overnight cultures grown in microtiter plates with 0.5% (w/v) crystal violet. Blank contained only media. Biofilm forming ability corresponded to OD 595 . B. Autoaggregation of 33277, FLL476 and FLL476C’ corresponded to change in OD 600 monitored for about three hours after cells were washed and suspended in PBS. A representative sample is shown. C. Hemagglutination activities of ATCC 33277, FLL476 and FLL476C’ were assessed by serially diluting cells in PBS and incubating with sheep RBCs for 3 h at 4°C. Dilutions are listed above and last dilution showing matt formation was taken as the titer. The blank contained only media. D. Antibiotic Protection Assay was used to quantify invasion. P. gingivalis cells that were able to invade HeLa cell monolayers were released by lysis and cultured on BA plates. Infectivity was taken as the percentage of cells recovered. (* = p

    Journal: PLoS ONE

    Article Title: In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose

    doi: 10.1371/journal.pone.0063367

    Figure Lengend Snippet: Comparison of biofilm formation, autoaggregation, hemagglutination and invasion assay. A. Biofilm formation of ATCC 33277, FLL476 and FLL476C’ were compared. Biofilm assay was done by staining adherent cells of overnight cultures grown in microtiter plates with 0.5% (w/v) crystal violet. Blank contained only media. Biofilm forming ability corresponded to OD 595 . B. Autoaggregation of 33277, FLL476 and FLL476C’ corresponded to change in OD 600 monitored for about three hours after cells were washed and suspended in PBS. A representative sample is shown. C. Hemagglutination activities of ATCC 33277, FLL476 and FLL476C’ were assessed by serially diluting cells in PBS and incubating with sheep RBCs for 3 h at 4°C. Dilutions are listed above and last dilution showing matt formation was taken as the titer. The blank contained only media. D. Antibiotic Protection Assay was used to quantify invasion. P. gingivalis cells that were able to invade HeLa cell monolayers were released by lysis and cultured on BA plates. Infectivity was taken as the percentage of cells recovered. (* = p

    Article Snippet: The vimF -defective mutant (FLL476) in the P. gingivalis ATCC 33277 genetic background showed a phenotype similar to that of the vimF -defective mutant (FLL95) in the P. gingivalis W83 genetic background.

    Techniques: Invasion Assay, Biofilm Production Assay, Staining, Lysis, Cell Culture, Infection

    Comparison of growth and gingipain activities of wild-type, vimF mutant and complemented strains of W83 and ATCC 33277. Growth rate of P. gingivalis ATCC 33277 ( A ) and W83 ( C ) were compared with their respective vimF -defective isogenic mutants (FLL476 and FLL95 ) and complemented strains (FLL476C’ and FLL95C’). The data shown is an average of three independent replicates. Error bars represent the SD. Gingipain activity of W83 ( D ) and ATCC 33277 ( B ) were compared with respective mutants and complemented strains. The activities were normalized to W83 and ATCC 33277 being 100% and the mutants reported as a percentage thereof. Error bars represent SD.

    Journal: PLoS ONE

    Article Title: In Porphyromonas gingivalis VimF Is Involved in Gingipain Maturation through the Transfer of Galactose

    doi: 10.1371/journal.pone.0063367

    Figure Lengend Snippet: Comparison of growth and gingipain activities of wild-type, vimF mutant and complemented strains of W83 and ATCC 33277. Growth rate of P. gingivalis ATCC 33277 ( A ) and W83 ( C ) were compared with their respective vimF -defective isogenic mutants (FLL476 and FLL95 ) and complemented strains (FLL476C’ and FLL95C’). The data shown is an average of three independent replicates. Error bars represent the SD. Gingipain activity of W83 ( D ) and ATCC 33277 ( B ) were compared with respective mutants and complemented strains. The activities were normalized to W83 and ATCC 33277 being 100% and the mutants reported as a percentage thereof. Error bars represent SD.

    Article Snippet: The vimF -defective mutant (FLL476) in the P. gingivalis ATCC 33277 genetic background showed a phenotype similar to that of the vimF -defective mutant (FLL95) in the P. gingivalis W83 genetic background.

    Techniques: Mutagenesis, Activity Assay

    Differential gene expression by comparative microarray analyses (represented in log 10 ) when comparing planktonic Porphyromonas gingivalis ATCC 33277 cells either in presence of a growing biofilm or in absence of a biofilm. Control planktonic cell gene expression (X-axis) is plotted against test cells (Y-axis) with a 1.5 fold change (up or down) and p -value

    Journal: BMC Microbiology

    Article Title: Comparative gene expression analysis of planktonic Porphyromonas gingivalis ATCC 33277 in the presence of a growing biofilm versus planktonic cells

    doi: 10.1186/s12866-019-1423-9

    Figure Lengend Snippet: Differential gene expression by comparative microarray analyses (represented in log 10 ) when comparing planktonic Porphyromonas gingivalis ATCC 33277 cells either in presence of a growing biofilm or in absence of a biofilm. Control planktonic cell gene expression (X-axis) is plotted against test cells (Y-axis) with a 1.5 fold change (up or down) and p -value

    Article Snippet: Slides specific for the strain P. gingivalis ATCC 33277 [Agilent Oligo Microarrays 8x15K (074976)] were used.

    Techniques: Expressing, Microarray

    Overview of experimental design. Porphyromonas gingivalis ATCC 33277 strain was maintained on blood agar plates and grown in modified Brain Heart Infusion (BHI) medium. After 24 h, optical density was measured, and a pure culture containing 10 8 colony forming units per milliliter (CFU/mL) was set. Two culture conditions were then prepared: Test cells, depositing P. gingivalis cells in presence of Hydroxyapatite (HA) disc, and Control cells, depositing P. gingivalis cells in the wells without HA discs. After 96 h of incubation of multi-well plates under anaerobic conditions, free floating P. gingivalis cells from both test and control condition were harvested, examined by CLSM, processed and total RNA extracted and purified. Agilent Oligo Microarrays 8x15K (074976) for P. gingivalis ATCC 33277 were used for hybridizations, (this slides also contents probes against the whole genome of P. gingivalis W83), and RT-qPCR analyses were performed to confirm the results. The experiments were repeated three times and each experimental condition was pooled into the three biological replicates and processed for the transcriptomic analysis. (Images for Fig. 1 were taken from https://smart.servier.com/ under a creative commons licence)

    Journal: BMC Microbiology

    Article Title: Comparative gene expression analysis of planktonic Porphyromonas gingivalis ATCC 33277 in the presence of a growing biofilm versus planktonic cells

    doi: 10.1186/s12866-019-1423-9

    Figure Lengend Snippet: Overview of experimental design. Porphyromonas gingivalis ATCC 33277 strain was maintained on blood agar plates and grown in modified Brain Heart Infusion (BHI) medium. After 24 h, optical density was measured, and a pure culture containing 10 8 colony forming units per milliliter (CFU/mL) was set. Two culture conditions were then prepared: Test cells, depositing P. gingivalis cells in presence of Hydroxyapatite (HA) disc, and Control cells, depositing P. gingivalis cells in the wells without HA discs. After 96 h of incubation of multi-well plates under anaerobic conditions, free floating P. gingivalis cells from both test and control condition were harvested, examined by CLSM, processed and total RNA extracted and purified. Agilent Oligo Microarrays 8x15K (074976) for P. gingivalis ATCC 33277 were used for hybridizations, (this slides also contents probes against the whole genome of P. gingivalis W83), and RT-qPCR analyses were performed to confirm the results. The experiments were repeated three times and each experimental condition was pooled into the three biological replicates and processed for the transcriptomic analysis. (Images for Fig. 1 were taken from https://smart.servier.com/ under a creative commons licence)

    Article Snippet: Slides specific for the strain P. gingivalis ATCC 33277 [Agilent Oligo Microarrays 8x15K (074976)] were used.

    Techniques: Modification, Incubation, Confocal Laser Scanning Microscopy, Purification, Quantitative RT-PCR

    Citrullinating activity in cell cultures. Activity of P. gingivalis strains containing variants PPAD‐T1 (ATCC 33277), PPAD‐T2 (ATCC T2; strain ATCC 33277 after introducing mutations G 231 N, E 232 T, and N 235 D), and a control, in which the antibiotic cassette used in ATCC T2 was introduced in wild‐type ATCC 33277 (ATCC T1 Ctrl). The results are shown as relative activity determined in triplicates using three independent cultures of each strain and displayed as mean ± SD. The statistical difference between strains was analyzed by the analysis of variance; ns, not significant; *** P

    Journal: Protein Science : A Publication of the Protein Society

    Article Title: Structure, function, and inhibition of a genomic/clinical variant of Porphyromonas gingivalis peptidylarginine deiminase

    doi: 10.1002/pro.3571

    Figure Lengend Snippet: Citrullinating activity in cell cultures. Activity of P. gingivalis strains containing variants PPAD‐T1 (ATCC 33277), PPAD‐T2 (ATCC T2; strain ATCC 33277 after introducing mutations G 231 N, E 232 T, and N 235 D), and a control, in which the antibiotic cassette used in ATCC T2 was introduced in wild‐type ATCC 33277 (ATCC T1 Ctrl). The results are shown as relative activity determined in triplicates using three independent cultures of each strain and displayed as mean ± SD. The statistical difference between strains was analyzed by the analysis of variance; ns, not significant; *** P

    Article Snippet: Along this line, the recent structure of PPAD from P. gingivalis reference strain ATCC 33277, hereafter PPAD‐T1, revealed the general architecture and modus operandi of this unique bacterial PAD., Moreover, recent studies reported that PPAD is present as point‐mutant variants differing from PPAD‐T1.

    Techniques: Activity Assay

    Fimbriae are not involved in P. gingivalis -mediated regulation of Angpt1 and Angpt2 production in AoSMCs. Quantitative real-time PCR results demonstrate relative transcription levels for Angpt1 (A) and Angpt2 (B) in AoSMCs stimulated with wild-type P.

    Journal: Infection and Immunity

    Article Title: Gingipains from the Periodontal Pathogen Porphyromonas gingivalis Play a Significant Role in Regulation of Angiopoietin 1 and Angiopoietin 2 in Human Aortic Smooth Muscle Cells

    doi: 10.1128/IAI.00498-15

    Figure Lengend Snippet: Fimbriae are not involved in P. gingivalis -mediated regulation of Angpt1 and Angpt2 production in AoSMCs. Quantitative real-time PCR results demonstrate relative transcription levels for Angpt1 (A) and Angpt2 (B) in AoSMCs stimulated with wild-type P.

    Article Snippet: The wild-type P. gingivalis strains ATCC 33277 and W50 significantly increased ETS1 expression in AoSMCs after 16 and 24 h ( ).

    Techniques: Real-time Polymerase Chain Reaction

    P. gingivalis and its gingipains and fimbrial mutants upregulate ETS1 in AoSMCs. Quantitative real-time PCR results demonstrate relative transcription levels of ETS1 in AoSMCs stimulated with wild-type ATCC 33277 and W50 and the corresponding W50 gingipain

    Journal: Infection and Immunity

    Article Title: Gingipains from the Periodontal Pathogen Porphyromonas gingivalis Play a Significant Role in Regulation of Angiopoietin 1 and Angiopoietin 2 in Human Aortic Smooth Muscle Cells

    doi: 10.1128/IAI.00498-15

    Figure Lengend Snippet: P. gingivalis and its gingipains and fimbrial mutants upregulate ETS1 in AoSMCs. Quantitative real-time PCR results demonstrate relative transcription levels of ETS1 in AoSMCs stimulated with wild-type ATCC 33277 and W50 and the corresponding W50 gingipain

    Article Snippet: The wild-type P. gingivalis strains ATCC 33277 and W50 significantly increased ETS1 expression in AoSMCs after 16 and 24 h ( ).

    Techniques: Real-time Polymerase Chain Reaction

    P. gingivalis and its gingipains regulate Angpt1 and Angpt2 expression in AoSMCs. (A and B) Quantitative real-time PCR results demonstrate relative transcription levels for Angpt1 (A) and Angpt2 (B) in AoSMCs stimulated with wild-type P. gingivalis (ATCC

    Journal: Infection and Immunity

    Article Title: Gingipains from the Periodontal Pathogen Porphyromonas gingivalis Play a Significant Role in Regulation of Angiopoietin 1 and Angiopoietin 2 in Human Aortic Smooth Muscle Cells

    doi: 10.1128/IAI.00498-15

    Figure Lengend Snippet: P. gingivalis and its gingipains regulate Angpt1 and Angpt2 expression in AoSMCs. (A and B) Quantitative real-time PCR results demonstrate relative transcription levels for Angpt1 (A) and Angpt2 (B) in AoSMCs stimulated with wild-type P. gingivalis (ATCC

    Article Snippet: The wild-type P. gingivalis strains ATCC 33277 and W50 significantly increased ETS1 expression in AoSMCs after 16 and 24 h ( ).

    Techniques: Expressing, Real-time Polymerase Chain Reaction