Journal: Nucleic Acids Research

Article Title: A WYL transcriptional regulator activates the DNA damage response pathway in Acinetobacter species

doi: 10.1093/nar/gkag250

Figure Lengend Snippet: A C-terminal 3xFLAG tag constitutively activates AciT. ( A ) Representative microscopy images of indicated strains. Scale bars, 10 µm. ( B ) Representative western blot on indicated strains grown with or without 2 µg/ml MMC. Blot was probed with ⍺-RpoA antibody as a loading control and ⍺-FLAG antibody to probe for AciT expression. ( C ) Growth curves of indicated strains grown with indicated concentrations of vancomycin in 96-well plates over 12 h. Arrows indicate concentration used for selection experiments. Each dot shows the mean of three independent, biological replicates, and error bars show SEM.

Article Snippet: Then, 1 μl of overnight cultures were subcultured into 200 μl fresh LB medium with vancomycin (MP Biomedicals) (0 μg/ml, 25 μg/ml, 50 μg/ml, 100 μg/ml, 250 μg/ml) grown in a 96-well plate in a Synergy H1 multimode plate reader (BioTek) utilizing Gen 6 software at 30°C shaking for 16 h. OD 600 was measured every 10 min.

Techniques: Microscopy, Western Blot, Control, Expressing, Concentration Assay, Selection