Fig 2From: Development of Pgrac100-based expression vectors allowing high protein production levels in Bacillus subtilis and relatively low basal expression in Escherichia coli Expression of BgaB in E. coli OmniMax and B. subtilis 1012 on X-gal plates and in liquid medium. a, Bacterial cells containing pHT01 (Pgrac, negative control), pHT01-bgaB (Pgrac01-bgaB), pHT100 (Pgrac100-bgaB) and pHT212 (Pgrac212-bgaB) were spotted on X-gal LB agar plates containing appropriate antibiotics and 0.01 mM IPTG for B. subtilis and without IPTG for E. coli at 30 °C for 48 h. Then, pictures were taken and single colonies are shown. b, The bacterial cells were grown in liquid LB medium at 37 °C to the mid-logarithmic growth phase, and then induced with 0.01 mM IPTG for B. subtilis and kept un-induced for E. coli. The cells were collected after 4 h of induction, and the BgaB activities were measured. The ratio of β-galactosidase activities of the samples were calculated from induced B. subtilis cells and un-induced E. coli cells. The ratio was set as one when the BgaB activities from both E. coli and B. subtilis were identical [4, 5]. c, E. coli cells containing pHT01 (Pgrac, negative control), pHT100 (Pgrac100-bgaB), pHCMC04-bgaB (PxylA-bgaB), pHCMC05-bgaB, pHT01-bgaB (Pgrac01-bgaB were spotted on X-gal LB agar plates containing ampicillin. d, the E. coli cells were grown in liquid LB medium at 37 °C to the mid-logarithmic growth phase, then the growing cells collected and the BgaB activities were measured. The ratio of β-galactosidase activities of the samples were calculated from different constructs to Pgrac01-bgaB Back to article page