Journal: AIChE journal. American Institute of Chemical Engineers
Article Title: Engineering Escherichia coli to produce and secrete colicins for rapid and selective biofilm cell killing.
doi: 10.1002/aic.17466
Figure Lengend Snippet: FIGURE 1 Biofilm cell killing with cell-free produced colicins. Time-course of killing of Escherichia coli K361 biofilm cells with the addition of cell-free produced colicins E1, E2, and E3. Overnight K361 cultures were adjusted to an OD600 nm of 0.01 (approximately 5 106 CFU/ml) in LB medium, incubated at 37C for 24 h to form biofilms, and exposed to each cell-free produced colicin (3.5 nM) for 5 min, 1 h, 4 h, and 8 h at 37C. Viable biofilm cells are quantified in colony-forming units per area (CFU/cm2). Error bars indicate the standard deviation from two independent cultures with three replicates
Article Snippet: AmpR, KanR, and StrR indicate ampicillin-, kanamycin-, and streptomycin-resistant, respectively Strains and plasmids Genotype/relevant characteristics Source Strains E. coli K361 W3110 strain, StrR 16 E. coli BL21Star (DE3) F ompT, hsdSB (rB mB ), gal, dcm, rne131 (DE3) Invitrogen E. coli K964 K91 containing pColE2 17 E. coli TG1 Strain containing colicin plasmid 18 E. coli DH5α fhuA2, Δ(argF-lacZ)U169, phoA, glnV44, Φ80 Δ(lacZ) M15, gyrA96, recA1, relA1, endA1, thi-1, hsdR17 NEB E. coli Nissle 1917 ΔdegP Probiotic E. coli (EcN) lacking degP, KanR 19 Lactococcus lactis 11454 Probiotic Lactococcus lactis subsp. Lactis ATCC strain (ATCC® 11454TM) ATCC VSL#3 Probiotic mixture VSL3 Pharma Plasmids pKSJ331 AmpR, ColE1 operon 18 pKSJ167 AmpR, ColE3 operon 20 pJL1-sfgfp KmR, PT7::sfGFP, C-terminal Strep-tag, pY71-sfGFP 21 pJL1-E1 KmR, PT7::colicin E1 and E1 immunity This study pJL1-E2 KmR, PT7::colicin E2 and E2 immunity This study pJL1-E3 KmR, PT7::colicin E3 and E3 immunity This study pBAD-gfp AmpR, ParaBAD::GFP Addgene (#54764) pBAD-empty AmpR This study pBAD-E2 AmpR, ParaBAD::colicin E2 and E2 immunity This study pBAD-E2+lys AmpR, ParaBAD::colicin E2 and E2 immunity, Ptet::E7 lysis, PproB::tetR This study nloaded from https://aiche.onlinelibrary.w iley.com /doi/10.1002/aic.17466 by E gyptian N ational Sti.
Techniques: Produced, Incubation, Standard Deviation
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