Fig. 1
Overproduction of VP1GFP in mutant strains defective in deacetylation (ΔcobB) or acetylation (ΔackA-pta). Bacteria were grown at 37 °C in LB medium supplemented with 100 µg/ml ampicillin. 1 mM IPTG was added at an OD595 = 0.5 to induce the synthesis of VP1GFP. a After 3, 6, and 24 h, whole-cell extracts were resolved by SDS-PAGE, subjected to immunodetection using anti-GFP antibodies and analyzed by densitometry to estimate the relative level of VP1GFP. b Representative growth curves of wt, ΔcobB, and ΔackA-pta cells. c GFP fluorescence in whole-cell extracts was recorded after 3 h of induction using EnSpire plate reader (PerkinElmer). Specific fluorescence emission of VP1GFP was estimated as described in the “Methods” section. d The amounts of VP1GFP in IBs were calculated in relation to the total VP1GFP level in bacterial extracts (100%). IBs were isolated from E. coli cells after induction of VP1GFP synthesis by 1 mM IPTG at the times indicated in the figure. Error bars represent the standard deviation of three values. AU-arbitrary units