non invasive e coli o157 strains  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability

doi: 10.3390/ijms25179224

Figure Lengend Snippet: Structure and analysis of E. coli O157 LPS molecules. ( A ) Schematic representation of E. coli O157 LPS molecule, showing O-antigen, outer core, inner core, and lipid A. Dotted lines indicate the level of LPS truncation resulting from each mutation (Kdo, 3-deoxy-D-manno-octulosonic acid; PPEtN, pyrophosphorylethanolamine; Hep, heptose; GlcNAc, N-acetylglucosamine; Glc, glucose; Gal, galactose). ( B ) LPS was isolated from wild-type and mutant cells of E. coli O157 43895 and analyzed by silver-stained Tricine–SDS-PAGE, as described in the Materials and Methods section. The expected location of O-antigen regions and the core is indicated on the left side of the gel.

Article Snippet: Unlike non-invasive E. coli O157 strains, ATCC 43895 forms a robust pellicle biofilm at 37 °C.

Techniques: Mutagenesis, Isolation, Staining, SDS Page

Journal: International Journal of Molecular Sciences

Article Title: Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability

doi: 10.3390/ijms25179224

Figure Lengend Snippet: Biofilm formation of E. coli O157 43895 WT and mutant derivatives. ( A ) Pellicle formation at 37 °C. Pellicle rings of a 48 h old floating biofilm of E. coli O157 43895. Mutants were detected by crystal violet (CV) staining. ( B ) Biofilm measurement using a standard microtiter assay. E. coli O157 43895 WT and mutants were incubated at 37 °C in MSM with 4% glucose. Bound CV was solubilized with 95% ethanol and quantified by absorbance at 595 nm. Assays were performed six times for each strain. Mean values with standard deviation are shown. Statistical analysis used one-way ANOVA test and Dunnett post-test. * p < 0.01, ** p < 0.001, compared with the wild type (WT).

Article Snippet: Unlike non-invasive E. coli O157 strains, ATCC 43895 forms a robust pellicle biofilm at 37 °C.

Techniques: Mutagenesis, Staining, Incubation, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability

doi: 10.3390/ijms25179224

Figure Lengend Snippet: The effect of LPS-core truncation on the swimming motility and flagella biosynthesis of E. coli O157 43895. Swimming motility of wild-type and LPS mutants was assessed using soft-agar swarm plates. After 40 h incubation, the diameter of the motility halo was measured. Experiments were repeated three times. Presence of flagella was detected by immunoblot (lower panel). Mean values with standard deviation are shown. Statistical analysis used two-tailed unpaired t -test. * p < 0.01, ** p < 0.001, compared with the wild type (WT).

Article Snippet: Unlike non-invasive E. coli O157 strains, ATCC 43895 forms a robust pellicle biofilm at 37 °C.

Techniques: Incubation, Western Blot, Standard Deviation, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability

doi: 10.3390/ijms25179224

Figure Lengend Snippet: The effect of LPS-core truncation on the curli biosynthesis of E. coli O157 43895. E. coli O157 43895 and mutants were grown on Congo-red (CR) indicator plates at 37 °C for 24 h. Presence of curli on the cell surface was detected by CR binding.

Article Snippet: Unlike non-invasive E. coli O157 strains, ATCC 43895 forms a robust pellicle biofilm at 37 °C.

Techniques: Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability

doi: 10.3390/ijms25179224

Figure Lengend Snippet: The effect of LPS core-OS truncation on cell adherence and invasion. ( A ) The adherence of LPS-core and flagella mutants of E. coli O157 43895 to epithelial cells was examined. The monolayers of MAC-T cells on coverslips in 24-well plates were co-cultured with the bacteria at 37 °C for 3 h. The cells were washed five times with PBS and fixed with 4% paraformaldehyde for 15 min, washed again with PBS, stained with Giemsa stain, and viewed under the microscope (magnification ×1000). ( B ) E. coli O157 43895 and the mutants were tested for epithelial-cell invasion using a MAC-T gentamicin protection assay, as described in the Materials and Methods section. Bacteria were co-cultured with MAC-T monolayers at a multiplicity of infection (MOI) of 10. The experiment was repeated three times with three replicates per strain, and CFUs were determined by plate count and were transformed to log10 value. Mean values with standard deviation are shown. Statistical analysis used one-way ANOVA test and Dunnett post-test. * p < 0.01, ** p < 0.001, compared with the wild type (WT).

Article Snippet: Unlike non-invasive E. coli O157 strains, ATCC 43895 forms a robust pellicle biofilm at 37 °C.

Techniques: Cell Culture, Bacteria, Staining, Giemsa Stain, Microscopy, Infection, Transformation Assay, Standard Deviation

Journal: International Journal of Molecular Sciences

Article Title: Lipopolysaccharide Core Truncation in Invasive Escherichia coli O157:H7 ATCC 43895 Impairs Flagella and Curli Biosynthesis and Reduces Cell Invasion Ability

doi: 10.3390/ijms25179224

Figure Lengend Snippet: E. coli O157 strains and plasmids.

Article Snippet: Unlike non-invasive E. coli O157 strains, ATCC 43895 forms a robust pellicle biofilm at 37 °C.

Techniques: Plasmid Preparation, Mutagenesis