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. 2019 Feb 15;47(7):e40. doi: 10.1093/nar/gkz072

Figure 6.

Figure 6.

Improve BLA expression in B. subtilis 6-102 by simultaneously activating molecular chaperone expression and repressing proteases expression by dCas9-ω. (A) Effects of activation of individual molecular chaperone and repression of individual protease on BLA production. GroES (intracellular molecular chaperone, Swiss-Prot: P28599), PrsA (extracellular molecular chaperone, Swiss-Prot: P24327) and six well-known extracellular protease genes (Bpr, bacillopeptidase F, Swiss-Prot: P16397; Vpr, extracellular serine protease, Swiss-Prot: P29141; NprB, extracellular neutral protease B, Swiss-Prot: P39899; Epr, extracellular serine protease, Swiss-Prot: P16396; WprA, cell wall-associated protein precursor, Swiss-Prot: P54423) were selected. (B) The relative expression of prsA, bpr, vpr and nprB measured by qPCR after expressing the dCas9-ω and sgRNAs. The strain 6-102/pHT01-dCas9-ω expressing sgRNA without target sequence was served as the control (C). (C) The synergistic effects of effective molecular chaperone and proteases on BLA production. The error bars indicate the standard deviations of biological triplicates.