e coli ec1000  (Addgene inc)

Journal: The Journal of Cell Biology

Article Title: Growth-dependent concentration gradient of the oscillating Min system in Escherichia coli

doi: 10.1083/jcb.202406107

Figure Lengend Snippet: MinD oscillation in cells of different lengths. (A) Time-lapse micrographs illustrating the pole-to-pole oscillation of sfGFP-MinD in cells of different lengths. The exponentially growing E. coli strain FW1541 was imaged at 12-s intervals for 10 min. Scale bar: 2 μm. (B) Standing wave features are demonstrated at 10% and 25% of the cell length, which was equivalent to the positions at 90% and 75%, respectively, along the medial axes of cells. (C) Kymographs demonstrating changes in the intensity profiles with length. (D) The standard deviation (SD) of the oscillation period is reduced in longer cells. (E and G) The correlation plots between the oscillation period and cell length when cultured with 0.416% (E) and 0% glucose (G). The oscillation periods range from 37 to 66 s for 0.416% glucose and from 41 to 99 s for 0% glucose. (F) Correlation between the oscillation velocity and the cell cycle duration. In A–C, MinD oscillations in three representative cells with varying lengths are presented. In D–G, analyses based on pooled data from multiple image fields are presented to demonstrate the oscillatory dynamics across cells of different lengths. Correlations were calculated using the nonparametric Spearman correlation method with 95% confidence intervals. The red dashed lines represent the lines fitted by simple linear regression. R 2 , goodness of fit; r, Spearman coefficient; P: two-tailed probability; n , population size. D–F, n = 130; G, n = 101.

Article Snippet: The E. coli strains MC1000 ( araD139 Δ(araABC-leu)7679 galU galK Δ(lac)X74 rpsL thi ; RRID: Addgene_71852) ( ) and DH5α ( ΔlacZ ΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17(rK-mK+) supE44 thi-1 gyrA96 relA1 ; RRDI: SCR_006368) ( ) were used for general cloning purposes.

Techniques: Standard Deviation, Cell Culture, Two Tailed Test

Journal: The Journal of Cell Biology

Article Title: Growth-dependent concentration gradient of the oscillating Min system in Escherichia coli

doi: 10.1083/jcb.202406107

Figure Lengend Snippet: Characteristic features of E. coli strains FW1541 and W3110 cultured in M9 medium supplemented with 0.416% glucose at 30°C. (A) Comparison of cell length, width, and aspect ratio. Data are presented as median values with interquartile ranges. Pooled data from several image fields were used for the analyses. Statistical comparisons were made using unpaired nonparametric Mann–Whitney tests. The median values are labeled and shown as the dashed lines, while the solid lines indicate the interquartile ranges. The P values of each pairwise comparison are labeled on top. W3110, n = 943; FW1541, n = 941. (B) Cell area (left) and aspect ratio (right), respectively, plotted against length and width. Correlations were calculated using the nonparametric Spearman correlation method with 95% confidence intervals. The same image dataset as in was used for the analysis. r, Spearman coefficient; P: two-tailed probability; n , population size. (C) A positive correlation between cell length and time was observed based on experimental data collected at 15-min intervals. The same image dataset as in , covering a complete cell cycle, was used for the analysis. Simple linear regression was applied to determine the length increase over time. The red line represents the best-fit regression line, while the blue dashed lines represent the errors. (D) A correlation plot of the period versus time was generated based on data from and . The Spearman nonparametric correlation method was used to calculate the correlation with 95% confidence intervals. The red dashed line represents the best-fit regression line. (B−D) R 2 , goodness of fit; r, Spearman coefficient; P: two-tailed; probability. n , population size. (E) Illustration of the cell volume calculation of FW1541.

Article Snippet: The E. coli strains MC1000 ( araD139 Δ(araABC-leu)7679 galU galK Δ(lac)X74 rpsL thi ; RRID: Addgene_71852) ( ) and DH5α ( ΔlacZ ΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17(rK-mK+) supE44 thi-1 gyrA96 relA1 ; RRDI: SCR_006368) ( ) were used for general cloning purposes.

Techniques: Cell Culture, Comparison, MANN-WHITNEY, Labeling, Two Tailed Test, Generated

Journal: The Journal of Cell Biology

Article Title: Growth-dependent concentration gradient of the oscillating Min system in Escherichia coli

doi: 10.1083/jcb.202406107

Figure Lengend Snippet: Examination of FtsA localization, which marks the division site, in relation to cell length and MinD oscillation period in E. coli strain FW1541. (A) Representative images demonstrating simultaneous observation of FtsA-mScarlet-I, sfGFP-MinD, and cell morphology. (B and C) The ring-like structure of FtsA-mScarlet-I had a relative position of 47.5 ± 2.1% from either pole (B), regardless of the cell length and the period of the MinD oscillations (C). The cell center is set at position 0.5 for normalization of the FtsA position. Pooled data from several image fields were used for the analysis. The Spearman nonparametric correlation method was used to calculate the correlation with 95% confidence intervals. The red dashed line represents the best-fit linear regression line. n , population size; R 2 , goodness of fit; r, Spearman coefficient; P: two-tailed probability.

Article Snippet: The E. coli strains MC1000 ( araD139 Δ(araABC-leu)7679 galU galK Δ(lac)X74 rpsL thi ; RRID: Addgene_71852) ( ) and DH5α ( ΔlacZ ΔM15 Δ(lacZYA-argF) U169 recA1 endA1 hsdR17(rK-mK+) supE44 thi-1 gyrA96 relA1 ; RRDI: SCR_006368) ( ) were used for general cloning purposes.

Techniques: Two Tailed Test

Journal: Cell

Article Title: Vaginal Lactobacillus fatty acid response mechanisms reveal a metabolite-targeted strategy for bacterial vaginosis treatment

doi: 10.1016/j.cell.2024.07.029

Figure Lengend Snippet:

Article Snippet: For plasmid construction, E. coli EC1000 strain (Addgene #71852), a kanamycin resistant strain carrying a single copy of the repA gene in the glgB gene, was used as the cloning host for all pORI28-based plasmids, and E. coli MC1061 strain (Molecular Cloning Laboratories #MC1061) was used as the cloning host for all pTRK892-based plasmids.

Techniques: Virus, Cloning, Molecular Cloning, Mutagenesis, Expressing, Plasmid Preparation, Recombinant, Electron Microscopy, Purification, Saline, Reverse Transcription, Clarification Assay, Viscosity, Gel Extraction, Transformation Assay, Electroporation, RNA Sequencing, Sequencing, Software, Transmission Assay, Microscopy