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Fig. 2 | Microbial Cell Factories

Fig. 2

From: Enhancing surfactin production by using systematic CRISPRi repression to screen amino acid biosynthesis genes in Bacillus subtilis

Fig. 2

Construction of the CRISPRi system and assay of the relative expression levels. a Schematic diagram of the integration of dCas9 and all sgRNA expression cassettes in the B. subtilis strain BS168NU-S. The dCaS9 cassette, which was expressed under the control of the inducible promoter of Pxyl, was integrated into the lacA site of the genome. The sgRNA was expressed by the constitutive promoter of Pveg and integrated into the amyE site of the genome. b Characterization of the expression levels of the inhibited genes through qRT-PCR at 24 h. The relative transcription level of the target gene was quantified by the 2−ΔΔCT method using the ccpA gene as the internal control and BS168NU-Sd as the calibrator

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