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. 2019 Aug 13;10(4):e01762-19. doi: 10.1128/mBio.01762-19

FIG 2.

FIG 2

Bfe1-based counterselection induced by aTC is highly effective. (A) Diagram of the two-step allelic replacement process. UHF, upstream homology flank; DHF, downstream homology flank. (B) aTC-inducible ssBfe1 counterselection cassette (53). (C) Vector pLGB13. bla, OriT, RK6, and MCS are described in the Fig. 1D legend. ermG, erythromycin resistance gene. (D) PCR verification of the counterselected clones for deletion of the 36-kb T6SS region of BfiCL09. WT, wild type; CI, cointegrate; C−, no-DNA control; MW, molecular weight marker. (Top) PCR using primers 1 and 2; (bottom) PCR using primers 3 and 4, as indicated in panel A.