Fig. 4
Verification of cre-1 deletion in selected transformants. a Schematic of homologous recombination (HR) of the target gene cre-1 mediated by Cas9, gRNA and donor DNA. (b-c) PCR analysis of selected transformants of M. thermophila b and M. heterothallica c with one primer (cre1-out-F) located upstream of the 5′ flanking region of the genomic DNA and the other (cre1-in-R) located in the 3′ flanking region of the genomic DNA. The expected length of disrupted transformants was 1.9 kb, while that of the host strain, used as a negative control, was 1.0 kb (rightmost lane)