Figure S6.
Effect of SPARTA on plasmid DNA and bacteriophages, related to Figure 6
(A) E. coli cells expressing MapSPARTA or MapSPARTATIR-E77A or harboring an empty BAC were transformed with pUC-empty or pUC-mRFPXylS-Pm, and the transformation efficiency was calculated. The average of three biological replicates is shown; error bars indicate standard deviations.
(B) E. coli cultures harboring pUC-empty and expressing MapSPARTA or MapSPARTATIR-E77A or harboring an empty BAC were grown without selective pressure (Ampicillin) to maintain pUC-empty. The fraction of cells containing pUC-empty was determined on a daily basis for nine consecutive days. The average of four biological replicates is shown; error bars indicate standard deviations.
(C) E. coli cultures expressing MapSPARTA or MapSPARTATIR-E77A or harboring an empty BAC were grown in agar plates in the presence of various bacteriophages, and the number of plaques relative to the strain harboring an empty BAC was determined. The average of three biological replicates is shown; data points indicate results from individual replicates.
(D) E. coli cultures expressing MapSPARTA or MapSPARTATIR-E77A or harboring an empty BAC were grown in the presence of various bacteriophages (bacteriophage type indicated above each graph), and the OD600 nm was monitored over time. Bacteriophages were added to E. coli cells with the following multiplicity of infection: T1—1.4 ∗ 10−7; T4—2 ∗ 10−7; T7—1 ∗ 10−7; Lambda—3 ∗ 10−7; Nami—1 ∗ 10−7. The average of three biological replicates is shown; shadings indicate standard deviations.
(E) E. coli JM109(DE3) cultures expressing MapSPARTA or MapSPARTATIR-E77A or harboring an empty vector were grown in the presence of chronic-infecting phage M13. The number of infectious M13 phage virions excreted into the bacterial culture was determined at multiple timepoints using E. coli ER2738 in which phage M13 is lytic. The average of three biological replicates is shown; error bars indicate standard deviations.