Bacterial Strains, Plasmids and Growth Media
Genotypes of the bacterial strains, plasmid descriptions, and sequences
of primers used in this study are listed in Tables S2-S3. Bacteria were
grown in DifcoTM LB Broth (BD #240230) consisting of
10 g/L tryptone, 5 g/L yeast extract, and 5 g/L sodium chloride. LB agar
plates were prepared from DifcoTM LB Agar (BD
#240110). The growth media was supplemented with an appropriate amount
of antibiotics (100 μg/mL ampicillin or carbenicillin, 25 μg/mL
chloramphenicol, 50 μg/mL kanamycin, 12.5 μg/mL tetracycline, 25-50
μg/mL zeocin) when necessary for the selection of strains and
maintenance of plasmids.
Keio collection strains, including ΔybeX , ΔybeY ,ΔybeZ , ΔrimM , ΔyjeQ and ΔksgA , andEscherichia coli wild-type BW25113 were used in this study (Babaet al. , 2006). We also reconstructed the ybeX single
deletion mutant using E. coli MG1655 and BW25113 via lambda red
recombination (Datsenko and Wanner, 2000). The kanamycin resistance gene
(kan) was removed from the bacterial chromosome using the pCP20 plasmid.
E. coli DH5α strain was used for plasmid cloning and propagation.
The TransBac library, an unpublished new E. coli overexpression
library based on a single-copy vector, was obtained from Dr Hirotada
Mori (Nara Institute of Science and Technology, Japan) as a stab stock
(Otsuka et al. , 2015).