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Published in final edited form as: Curr Genet. 2018 Jun 16;65(1):179–192. doi: 10.1007/s00294-018-0858-0

Fig. 3 — a. Fluorescence emission distribution in the presence of antibiotic, as an approximate indication of plasmid copy number distribution in the population. Cells transformed with WT pGFPuv (WT, grey line), DAS-pGFPuv (DAS, black line), pGFPuv DAS-G180A (DAS180A, blue line), or pGFPuv DAS-G512A (DAS512, red line), were grown in the presence carbenicillin and analyzed by flow cytometry. Fluorescence emission and scatter for 250,000 single cells was analyzed by flow cytometry as described in Methods. All distributions were centered in the geometric mean to allow comparison. Three subpopulations were defined based on the fluorescence emission profile as described in Methods. The results show one representative experiment. b. Plasmid retention. Cells carrying the WT pGFPuv plasmid (WT), the pGFPuv-DAS plasmid (DAS) or the pGFPuv-DAS plasmid with the G512A substitution (DAS_G512A) were grown in the presence of carbenicillin, analyzed by flow cytometry, and placed into one of the three categories following the parameters defined for the control: no plasmid (white), low plasmid (light grey), or high plasmid (dark grey). c. Same analysis performed after a single passage in the absence of carbenicillin as described in the legend for Fig. 2 and in Methods. d. Same analysis performed after four passages in the absence of carbenicillin as described in the legend for Fig. 2 and in Methods.

Fig. 3 — a. Fluorescence emission distribution in the presence of antibiotic, as an approximate indication of plasmid copy number distribution in the population. Cells transformed with WT pGFPuv (WT, grey line), DAS-pGFPuv (DAS, black line), pGFPuv DAS-G180A (DAS180A, blue line), or pGFPuv DAS-G512A (DAS512, red line), were grown in the presence carbenicillin and analyzed by flow cytometry. Fluorescence emission and scatter for 250,000 single cells was analyzed by flow cytometry as described in Methods. All distributions were centered in the geometric mean to allow comparison. Three subpopulations were defined based on the fluorescence emission profile as described in Methods. The results show one representative experiment. b. Plasmid retention. Cells carrying the WT pGFPuv plasmid (WT), the pGFPuv-DAS plasmid (DAS) or the pGFPuv-DAS plasmid with the G512A substitution (DAS_G512A) were grown in the presence of carbenicillin, analyzed by flow cytometry, and placed into one of the three categories following the parameters defined for the control: no plasmid (white), low plasmid (light grey), or high plasmid (dark grey). c. Same analysis performed after a single passage in the absence of carbenicillin as described in the legend for Fig. 2 and in Methods. d. Same analysis performed after four passages in the absence of carbenicillin as described in the legend for Fig. 2 and in Methods.