electrotransformation method  (ATCC)

Journal: Applied and Environmental Microbiology

Article Title: Mutations in Peptidoglycan Synthesis Gene ponA Improve Electrotransformation Efficiency of Corynebacterium glutamicum ATCC 13869

doi: 10.1128/AEM.02225-18

Figure Lengend Snippet: Electrotransformation efficiencies of the ATCC 13869 wild-type (WT) strain and mutants. Competent cells were prepared using LBG medium without cell wall-weakening agents. The number of cells used for each electrotransformation was approximately 108. Error bars indicate standard deviations from the results from three parallel experiments. Asterisks indicate significant changes in electrotransformation efficiency based on a comparison between the mutants and the wild-type strain. **, P ≤ 0.01; ***, P ≤ 0.001 (Student’s two-tailed t test).

Article Snippet: Since the ponA Y489C mutant showed the highest transformation efficiency, it was further studied in the subsequent experiments. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG 1 caption a7 Electrotransformation efficiencies of the ATCC 13869 wild-type (WT) strain and mutants.

Techniques: Comparison, Two Tailed Test

Journal: Applied and Environmental Microbiology

Article Title: Mutations in Peptidoglycan Synthesis Gene ponA Improve Electrotransformation Efficiency of Corynebacterium glutamicum ATCC 13869

doi: 10.1128/AEM.02225-18

Figure Lengend Snippet: Effects of Y489C point mutation and deletion of ponA on electrotransformation efficiency. (A) Function of PonA in transglycosylation and transpeptidation of cell wall synthesis. (B) Electrotransformation efficiency and susceptibility to electroporation of the wild-type (WT), ponAY489C, and ΔponA strains in the absence of cell wall-weakening agents. Competent cells were prepared using LBG medium without cell wall-weakening agents. The cells used for each electrotransformation were approximately 108. After electroporation and recovery, cells were spread on selective LBG plates supplemented with 25 µg/ml kanamycin to determine the electrotransformation efficiency and LBG plates without kanamycin to determine the susceptibility to electroporation. (C) Growth curves of wild-type (WT), ponAY489C, and ΔponA strains. Cells were cultured in LBG medium with an initial OD600 value of 0.2. The cell density was determined every 3 hours. (D) Electrotransformation efficiency of wild-type (WT), ponAY489C, and ΔponA strains in the presence of cell wall-weakening agents. Competent cells were prepared using NCM medium supplemented with glycine, threonine, INH and Tween 80 according to the protocol described previously (12). Error bars indicate standard deviations from three parallel experiments. Asterisks indicate significant changes in electrotransformation efficiency and susceptibility to electroporation based on a comparison between the mutants and the wild-type strain. **, P ≤ 0.01; ***, P ≤ 0.001 (Student’s two-tailed t test). NS indicates nonsignificant change between the wild-type strain and the mutant based on Student’s two-tailed t test (P > 0.05).

Article Snippet: Since the ponA Y489C mutant showed the highest transformation efficiency, it was further studied in the subsequent experiments. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window FIG 1 caption a7 Electrotransformation efficiencies of the ATCC 13869 wild-type (WT) strain and mutants.

Techniques: Mutagenesis, Electroporation, Cell Culture, Comparison, Two Tailed Test