MOESM4 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 4: Figure S3. (A) Sequence alignment of three alanine racemases from S. Enteritidis (Alr, DadX, SEN3897). Alr (SEN4016) showed 91% with Y274f Alanine Racemase from E. Coli (PDB-ID-4WR3), DadX (SEN1235) showed 48% identity with Alanine Racemase from Pseudomonas aeruginosa, (PDB-ID-1RCQ) and SEN3897 showed 42% identity with Chain A, Alanine Racemase from Bartonella henselae (PDB-ID-3KW3). (B) Ribbon protein structures determined by I-TASSER. Superimposed alanine racemase structures of Alr (green), DadX (red) and SEN3897 (turquoise Blue) denote their structural similarities with residue conservation

Interscientific Integration is in the Comprehension of Mnogourovnevosti Sociokul'turnikh Regulyativov of Human Activity

У статті окреслено основні історичні ступені дослідження сфери регуляції людської діяльності засобами природознавства, неврології, психології. Доведено, що існує специфічний напрям знання, в якому втілюються різноманітні методологічні підходи, парадигми, інструментальні технології вивчення внутрішніх механізмів регуляції людської діяльності.The basic historical stages of research of sphere of regulation of human activity are marked by facilities of natural science, neurology, psychology. It is well-proven that exists specific direction of knowledge, various metodzhology approaches, paradigms, instrumental technologies of study of internal mechanisms of regulation of human activity, will be realized in which

MOESM10 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 10: Figure S8. Colonization characteristics of SEN ∆alr, SEN ∆dadX, SEN ∆3897, SEN ∆alr∆dadX and WT: Feces from each group of 5 mice were collected and suitable dilutions (1:200, 1:40,000) were plated on MacConkey agar plates with required antibiotics at 24 h and 48 h p.i for assessing the bacterial counts. Statistical significance: *P < 0.05, **P < 0.01

MOESM6 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 6: Figure S4. Expression of SEN3897 in WT and SEN ∆alr∆dadX bacterial strains through qRT-PCR analysis. All experiments were performed in triplicates with data represented as mean ± SD. Statistical significance: **P < 0.01

MOESM9 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 9: Figure S7. Fluorescence microscopy images of murine macrophages RAW264.7 infected with SEN ∆alr, SEN ∆dadX, SEN ∆3897 and WT for 1 h at MOI 10, gentamicin treated for 24 h and fixed. Cell nuclei were stained with DAPI (blue) and Salmonella were GFP-tagged (green). Scale bar: 10 μm

MOESM7 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 7: Figure S5. Adhesion assay of SEN ∆alr, SEN∆dadX, SEN ∆3897 and WT in HCT116 cell-lines for 30 min at MOI 10. Statistical significance: *P < 0.05

MOESM2 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 2: Figure S1. PCR screening of alanine racemase gene inactivation. (A) Confirmation of PCR mediated gene replacement of alr, dadX and SEN ∆3897 in Salmonella Enteritidis by kanamycin cassette with internal kanamycin specific primers and confirmatory primers (B) PCR confirmation of double deletions (SEN∆alr∆dadX) in S. Enteritidis by phage transduction with FRT specific primers for kanamycin and chloramphenicol cassettes of 1.5 kb and 1.1 kb size respectively. These ethidium bromide stained gels were analyzed by agarose gel electrophoresis. The 1 kb DNA ladder (Invitrogen) with the desired product sizes is indicated with arrows

MOESM1 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 1: Table S1. Primer list used in this study

MOESM5 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 5: Table S2. Active site residues of alanine racemase proteins (Alr, DadX and SEN3897) of Salmonella Enteritidis from CASTp analysis

MOESM11 of Identification of a new alanine racemase in Salmonella Enteritidis and its contribution to pathogenesis

Additional file 11: Figure S9. (A) Invasion assay of SEN ∆alr∆3897 and WT in HCT116 cell-lines. SEN ∆invC acts as the negative control for invasion assay (B) Uptake Assay and (C) Survival Assay of SEN ∆alr∆3897 and WT in RAW264.7 in murine macrophages at 2 h and 24 h time-points respectively. Statistical significance: *P < 0.05, ****P < 0.0001