Journal: Frontiers in Microbiology

Article Title: Budding and explosive membrane vesicle production by hypervesiculating Escherichia coli strain Δ rodZ

doi: 10.3389/fmicb.2024.1400434

Figure Lengend Snippet: Escherichia coli strains and plasmids used in this study.

Article Snippet: The WT E. coli K-12 strain BW25113 and rodZ -deficient mutant were obtained from the National BioResource Project (National Institute of Genetics [NIG], Mishima, Japan) ( ).

Techniques: Cloning, Plasmid Preparation

Journal: Frontiers in Microbiology

Article Title: Budding and explosive membrane vesicle production by hypervesiculating Escherichia coli strain Δ rodZ

doi: 10.3389/fmicb.2024.1400434

Figure Lengend Snippet: Relative mreB expression in Escherichia coli strains with CRISPRi system. The expression level was normalized against that in wild-type (WT) cells. The vertical bars indicate standard deviations (calculated from more than three independent experiments). Statistically significant difference from the WT strain ( p < 0.05) is marked with asterisk.

Article Snippet: The WT E. coli K-12 strain BW25113 and rodZ -deficient mutant were obtained from the National BioResource Project (National Institute of Genetics [NIG], Mishima, Japan) ( ).

Techniques: Expressing

Journal: Frontiers in Microbiology

Article Title: Budding and explosive membrane vesicle production by hypervesiculating Escherichia coli strain Δ rodZ

doi: 10.3389/fmicb.2024.1400434

Figure Lengend Snippet: Cell growth and relative outer membrane vesicle (OMV) production by each E. coli strain. (A) OD 660 values of culture of each E. coli strain after 24 h. (B) Relative OMV production by each E. coli strain, evaluated by staining with FM4-64 dye. (C) Relative OMV production by each E. coli strain, evaluated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). OMV production was normalized to that by the WT strain. Vertical bars indicate standard deviations (calculated from more than three independent experiments). Statistically significant differences from the WT strain ( p < 0.05) are marked with asterisks.

Article Snippet: The WT E. coli K-12 strain BW25113 and rodZ -deficient mutant were obtained from the National BioResource Project (National Institute of Genetics [NIG], Mishima, Japan) ( ).

Techniques: Membrane, Staining, Polyacrylamide Gel Electrophoresis, SDS Page

Journal: Frontiers in Microbiology

Article Title: Budding and explosive membrane vesicle production by hypervesiculating Escherichia coli strain Δ rodZ

doi: 10.3389/fmicb.2024.1400434

Figure Lengend Snippet: Surface structure of the E. coli WT, Δ rodZ , Δ rodZ + prodZ (complementary strain), and mreB R3 (expression of mreB repressed—see ) cells visualized via quick-freeze, deep-etch electron microscopy (QFDE-EM). Field image of WT (A) , Δ rodZ (C) , Δ rodZ + prodZ (E) , and mreB R3 (G) cells. Magnified image of the surface of WT (B) , Δ rodZ (D) , Δ rodZ + prodZ (F) , and mreB R3 (H) cells. The cells were collected after 24 h of culture.

Article Snippet: The WT E. coli K-12 strain BW25113 and rodZ -deficient mutant were obtained from the National BioResource Project (National Institute of Genetics [NIG], Mishima, Japan) ( ).

Techniques: Expressing, Electron Microscopy

Journal: Frontiers in Microbiology

Article Title: Budding and explosive membrane vesicle production by hypervesiculating Escherichia coli strain Δ rodZ

doi: 10.3389/fmicb.2024.1400434

Figure Lengend Snippet: Aberrant surface structures of E. coli Δ rodZ cells. (A,B) Cells with a budding surface. (C) Magnified image of the surface structure of the budding vesicles. The arrowheads indicate vesicles budding from the cell surface. (D) Cells with the budding vesicles and a dented surface. (E) Magnified image of a budding vesicle; the arrowhead indicates the budding vesicle. (F) Magnified image of a dented surface; the arrowheads indicate the dents on the surface. (G,H) Cells with the curved pattern on the surface. (I) Magnified image of a curved pattern. The arrowhead indicates the curved pattern, which randomly appeared on the cell surface.

Article Snippet: The WT E. coli K-12 strain BW25113 and rodZ -deficient mutant were obtained from the National BioResource Project (National Institute of Genetics [NIG], Mishima, Japan) ( ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Budding and explosive membrane vesicle production by hypervesiculating Escherichia coli strain Δ rodZ

doi: 10.3389/fmicb.2024.1400434

Figure Lengend Snippet: Cross-section of various fractured E. coli cells. Field image of the structure of WT (A) , Δ rodZ (C) , and mreB R3 (E) cells. Magnified image of the surface of WT (B) , Δ rodZ (D) , and mreB R3 (F) cells. The cells were collected after 24 h of culture.

Article Snippet: The WT E. coli K-12 strain BW25113 and rodZ -deficient mutant were obtained from the National BioResource Project (National Institute of Genetics [NIG], Mishima, Japan) ( ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Budding and explosive membrane vesicle production by hypervesiculating Escherichia coli strain Δ rodZ

doi: 10.3389/fmicb.2024.1400434

Figure Lengend Snippet: Distribution of cell volumes for each E. coli strain. The cells were harvested after 24 h of culture. Cell volume was determined using the qNano system.

Article Snippet: The WT E. coli K-12 strain BW25113 and rodZ -deficient mutant were obtained from the National BioResource Project (National Institute of Genetics [NIG], Mishima, Japan) ( ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Budding and explosive membrane vesicle production by hypervesiculating Escherichia coli strain Δ rodZ

doi: 10.3389/fmicb.2024.1400434

Figure Lengend Snippet: Model of the proposed mechanism underlying the enhanced production of vesicles by the E. coli Δ rodZ strain. The Δ rodZ cells become spherical and expanded by osmotic pressure due to their incomplete peptidoglycan (PG) structure; budding vesicles are generated from the sites of holes in the PG. Budding traces were also observed by QFDE-EM. In parallel, cell burst and the reassociation of membrane fragments result in explosive vesicle production. This model was supported by the restored OD 660 value and decreased vesicle production in conditions of osmotic support.

Article Snippet: The WT E. coli K-12 strain BW25113 and rodZ -deficient mutant were obtained from the National BioResource Project (National Institute of Genetics [NIG], Mishima, Japan) ( ).

Techniques: Generated, Membrane