Article Figures & Data
Figures
- FIG 1
IIL tolerance of selected bacterial strains. Growth curves (A) and maximum growth rate and maximum cell density (B) are shown for each strain cultured in IIL medium (250 mM [C2C1im][OAc]). The chemical structure of [C2C1im][OAc] is shown in the inset in panel A.
- FIG 2
Identification of genome regions containing IIL tolerance genes. (A) The top blue line depicts the B. cereus loci from which the IILT-fosmid inserts originated. Region 2 is expanded below. (B and C) As indicated by growth curves (B) and maximum growth rate and maximum cell density (C), E. coli is protected from IIL (250 mM [C2C1im]Cl) by both fosmids and pBbS0c plasmids (48) containing B. cereus DNA from these loci.
- FIG 3
IIL tolerance in B. thuringiensis depends upon tandem SMR pump genes. Growth curves (A) and maximum growth rate and maximum cell density (B) are shown for each strain cultured in IIL medium (360 mM [C2C1im][OAc]).
- FIG 4
Point mutations in the guanidine II riboswitch confer IIL tolerance to E. coli. (A) The highly conserved ACGR sequence within the P1 and P2 hairpins of the guanidine II riboswitch that regulates sugE in E. coli. (B and C) Growth curves (B) and maximum growth rate and maximum cell density (C) are shown for the plasmid expression of various sugE constructs that complement ΔsugE in E. coli grown in IIL medium (250 mM [C2C1im]Cl).
- FIG 5
Addition of guanidine protects E. coli from IIL toxicity. E. coli DH10b was cultured in medium containing either 0 or 10 mM guanidine-HCl; 250 mM [C2C1im][OAc] was added at 0, 30, and 180 min after the start of culture. (A) Growth curves, with the addition of IIL indicated by dashed lines; (B) maximum cell density at the initial IIL addition and after 20 h of cultivation.
- FIG 6
Efflux pump library tested in E. coli. Efflux pumps from the various organisms noted were expressed in E. coli DH10b in the IPTG-inducible vector pBbS6k. Growth curves (A) and maximum growth rate and maximum cell density (B) are shown for each strain cultured in medium containing 250 mM [C2C1im]Cl, 0.1 mM IPTG, and 50 mg/ml kanamycin.
Additional Files
Supplemental material
- Supplemental file 1 -
Fig. S1 (Restoration of IILS phenotype in a pump knockout strain of B. thuringiensis), S2 (B. licheniformis pump genes and IIL tolerance in E. coli), S3 (GFP expression), S4 (Rainforest metagenomic DNA and tolerance phenotypes in E. coli), and S5 (Plasmid expression of sugE in E. coli EPI300) and Tables S1 (Strains), S2 (Strains by figure), and S3 (Vectors)
PDF, 437K
- Supplemental file 1 -
