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. 2020 Dec 8;11(6):e03012-20. doi: 10.1128/mBio.03012-20

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Copyright © 2020 Park 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 6 — Rescue of FtsI mutants by activated FtsL and FtsW mutants. (A) Rescue of FtsI mutants by FtsW^M269I. To test if the FtsI mutants could be rescued by an activated allele of ftsW, MCI23 (ftsI23^ts recA::spc) was transformed with compatible plasmids expressing an activated allele of ftsW (pSEB429 [P₂₀₄::ftsW^M269I]) and ftsI alleles under arabinose promoter control (derivatives of pKTP109 [P_ara::ftsI]). Transformants were spot tested on plates at 37°C (to inactivate ftsI23^ts) with arabinose added to induce the ftsI alleles and increasing concentrations of IPTG to induce ftsW^M269I. Note: additional alleles of ftsI were not rescued by ftsW^M269I (Fig. S8). (B) Rescue of FtsI mutants by ftsL^E88K/G92D. To test rescue of FtsI mutants by activated FtsL, MCI23 (ftsI23^ts recA::spc) was transformed with compatible plasmids expressing an activated allele of ftsL (pKTP100* [P_tac::ftsL^E88K/G92D]) and the various ftsI alleles under arabinose promoter control (derivatives of pKTP109 [P_ara::ftsI]). Transformants were spot tested on plates at 37°C (to inactivate ftsI23^ts), and arabinose was added to induce the ftsI alleles, and increasing concentrations of IPTG were added to induce ftsL^E88K/G92D.