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. 2021 Aug 12;12:697712. doi: 10.3389/fmicb.2021.697712

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Copyright © 2021 Kasho, Oshima, Chumsakul, Nakamura, Fukamachi and Katayama.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Requirement for DnaA box R1 for oriC–IHF binding at the replication initiation period. (A) Deletion derivatives of oriC. Bars indicate the regions of oriCΔDUE and oriCΔ(DUE-R1), which were inserted at the position indicated as TER-oriC in the terC locus [shown in panel (B)] of SH022 (dnaC2) strain, resulting in strains KX237 and KX238. Symbols used for the oriC structure are shown as in Figure 1B. (C,D) IHF-ChAP-qPCR with TER-oriC-inserted cells. SH022 (dnaC2), KX237 (dnaC2 TER-oriCΔDUE), and KX238 [dnaC2 TER-oriCΔ(DUE-R1)] cells grown at 30°C (R; Random) were transferred to 38°C and incubated for 90 min (S; Synchronized). The relative levels of native oriC (C) and TER-oriC (D) before and after Ni-affinity purification were determined using real-time qPCR, and yield was calculated (expressed as %). Error bars represent standard deviation from three independent experiments.