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. 2021 Dec 8;6(6):e00919-21. doi: 10.1128/msphere.00919-21

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Copyright © 2021 Edwards et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 7 — PCR verification of the constructed strains. (A) PCR confirmation of ΔdccA in the R20291 and 630Δerm backgrounds using primer pair R2926/R2927. Shown are strains R20291, R20291 ΔdccA (RT2656), 630Δerm, and 630Δerm ΔdccA (RT2703). The expected PCR products’ sizes are 2.5 kb for the wild-type allele and 1.7 kb for the deletion mutants. (B) PCR verification of Tn916::P_CD1421-dccA chromosomal integration in 630Δerm and R20291 using the internal qRT-PCR dccA primer pair CD1420qF/CD1420qR. Shown are strains 630Δerm ΔdccA (RT2703), 630Δerm ΔdccA Tn916::P_CD1421-dccA (MC1960), R20291, R20291 ΔdccA (RT2656), and R20291 ΔdccA Tn916::P_CD1421-dccA (MC1961). The expected PCR product size is 140 bp. (C) PCR confirmation of the CD2492-targeted intron in the 630Δerm ΔCD1492 background (MC674). Shown are strains 630Δerm, 630Δerm ΔCD1492 (MC674), and 630Δerm ΔCD1492 CD2492::erm (MC802) using primer pair oMC309/oMC338. The expected PCR products’ sizes are 811 bp for the wild-type CD2492 allele and ∼2,811 bp for CD2492::erm.