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. 2019 Aug 14;85(17):e01076-19. doi: 10.1128/AEM.01076-19

FIG 4.

FIG 4

The effect of 5′ UTR mutation on the production of Dac by the recombinant B. subtilis WB600. (a) The extracellular Dac activities of the recombinant strains. (b) SDS-PAGE analysis of extracellular Dac of the recombinant strains. M, protein marker; 1, WB-NMK/Dac (mature Dac expressed in B. subtilis WB600 strains harboring p43NMK/Dac plasmid); 2, WB-NMKmut/Dac (mature Dac expressed in B. subtilis WB600 strains harboring p43NMKmut/Dac plasmid). (c) qRT-PCR analysis of the Dac mRNA in the recombinant strains WB-NMK/Dac and WB-NMKmut/Dac at different culture phases. The recombinant strains were grown in fermentation medium (see Materials and Methods) containing 10 mg/liter of kanamycin in 250-ml flask and incubated at 37°C and 220 rpm for 60 h. Error bars indicate the average values from three parallel experiments.