Plasmid miniprep kit

The strains and plasmids used in this study are listed in Table S1. E. coli strains 10G (Lucigen) and TOP10 (Invitrogen) were used for cloning and biosensor expression, respectively. Aeromicrobium erythreum NRRL B3381 and the corresponding knock-out strain were a kind gift from Prof. Eric Miller, NC State University. The plasmid pMLGFP was used as a template for MphR mutagenesis and to express the reporter gene in the biosensor strains. Bacteria were grown in Luria Broth supplemented with ampicillin and tetracycline as appropriate. ErA, clarithromycin, azithromycin, and roxithromycin were from Sigma-Aldrich and pikromycin was from Abcam. YC-17 was a kind gift from Prof. David Sherman, University of Michigan. Each macrolide was prepared in dimethyl sulfoxide (DMSO) to a stock concentration of 50 mM, 5 mM, 500 μM, or 50 μM. All other chemicals were purchased from Sigma-Aldrich unless stated otherwise. PCR products were extracted with a Bio Basic Gel Extraction Kit. Restriction enzymes were purchased from New England Biolabs. Plasmids were isolated using a plasmid miniprep kit from Bio Basic.

All other chemicals were purchased from Sigma-Aldrich unless stated otherwise. PCR products were extracted with a Bio Basic Gel Extraction Kit. Restriction enzymes were purchased from New England Biolabs. Plasmids were isolated using a plasmid miniprep kit from Bio Basic. Other DNA preparation kits (genomic, and gel extraction) were purchased from (NEB). Oligos were synthesized by Integrated DNA Technologies (IDT) and purified by IDT using standard desalting. Polymerases were purchased from Fisher Scientific; all restriction enzymes were purchased from New England Biolabs (NEB). LB media was purchased from Fisher Scientific. Polyacrylamide gels were homemade and prepared using reagents purchased from Fisher Scientific; gels were prepared using 4% acrylamide stacking gel and 20% acrylamide running gel. Dibenzocyclooctyne-fluor (DBCO) 488 reagent was purchased from Sigma Aldrich. Reagents used for buffer preparation were purchased from VWR. Isopropyl-β-D-thiogalactopyranoside (IPTG) was purchased from CalBioChem. Bacillus thuringiensis 4BD1 strain was ordered from the Bacillus Genetic Stock Center (BGSC). For protein expression, E. coli strain BL21(DE3) was used unless stated otherwise. For DNA storage and manipulation, E. coli TOP10 (Invitrogen) was used.

The strains and plasmids used in this study are listed in Supplementary Table S2. E. coli strains 10G (Lucigen) TOP10 (Invitrogen) were used for cloning and biosensor expression, respectively. Bacteria were grown in Luria Broth (Fisher Scientific) supplemented with ampicillin (Fisher Scientific) and tetracycline (Fisher Scientific) as appropriate. ErA and CLA were obtained from Sigma Aldrich. Each macrolide was prepared in dimethyl sulfoxide (DMSO) to a stock concentration of 0.5 μM, 5 μM, 50 μM, or 500 μM. DMSO and 96-deepwell plates were purchased from Fisher Scientific. All other chemicals were purchased from Sigma-Aldrich unless stated otherwise. PCR products were extracted with a Bio Basic Gel Extraction Kit. All enzymes for DNA manipulations were purchased from New England Biolabs. Plasmids were isolated using a plasmid miniprep kit from Bio Basic. All oligonucleotides were purchased from Integrated DNA Technologies. Clear and opaque flat-bottom 96-well plates were purchased from Greiner Bio-One. All other chemicals were reagent grade or better.