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

FIG 3.

FIG 3

Deletion of the tetracycline resistance gene tetQ. (A) Allelic replacement vector pLGB29, using inulin selection for cointegrates. bla, oriT, RK6, and MCS are described in the Fig. 1D legend. (B) PCR verification and growth on plates of the counterselected clones for the tetQ deletion in BoCL09. An inulin selection plate for the mating of BoCL09T03C03, using homology-based integration of pLGB29 containing 1 kb of DNA on each side of the tetQ gene, is shown. S, sensitive; R, resistant. (C, left) Ethidium bromide-stained agarose gel showing PCR amplicons of the resolvent for the tetQ deletion in PmCL09 using pLGB13 (erythromycin sensitive). (Right) PCR amplicon to verify excision of the whole CTnYCH46-1 (HMPREF1078_01847 to HMPREF1078_01893). Primers anneal upstream and downstream of the 49-kb conjugative transposon. WT, wild type; CI, cointegrate. C−, no-DNA control; MW, molecular weight marker.