Fig. 3

The genome editing efficiency of the all-in-one plasmid. a PCR verification of erm gene knockout. Lanes 1–32 represent the PCR product of mutant (1583 bp), respectively. All 32 clones were shown to harbor the desired deletion by colony PCR; CK represents the PCR product of wild-type strain (1949 bp). b PCR verification of amyE gene knockout. Lanes 1–32 represent the PCR product of mutant (1114 bp), respectively. All 32 clones were shown to harbor the desired deletion by colony PCR; CK represents the PCR product of wild-type strain (1614 bp). c Phenotypic verification of erm gene knockout. The wild-type strain grew well on the erythromycin plate, while △erm mutant strain was sensitive to erythromycin due to the knockout of the erythromycin gene, resulting in inhibited growth on the plate. d Phenotypic verification of amyE gene knockout. The wild-type strain has obvious hydrolysis ring, while the △amyE mutant strain displayed no transparent ring in the starch-plate. e Phenotypic verification of the point mutation of spo0A gene. The colony morphology of spo0A^A714T mutant strain was more transparent. f Sporulation germination rates of the wild-type strain BSSCK6 and the Spo0A point mutation strain BS02. g PCR verification of the egfp insertion efficiency. Lanes 1–32 represent the PCR product of mutant (2711 bp), respectively. CK represents the PCR product of wild-type strain (1614 bp)