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. Author manuscript; available in PMC: 2022 Aug 1.
Published in final edited form as: Mol Microbiol. 2021 Jun 25;116(2):707–722. doi: 10.1111/mmi.14765

Figure 1. yneA C-terminal truncations impair checkpoint activation.

Figure 1.

(A) Schematic of the full-length YneA protein and the C-terminal truncation mutants lacking the last five (yneAΔ5), ten (yneAΔ10) and fifteen (yneAΔ15) amino acid residues. YneA is predicted to have a transmembrane domain (TM) and a LysM binding domain (LysM). (B) Spot titer assay using B. subtilis strains WT (PY79), ΔyneA::loxP amyE::Phy-yneA (EAM46), ΔyneA::loxP amyE::Phy-yneAΔ5 (EAM53), ΔyneA::loxP amyE::Phy-yneAΔ10 (EAM54) and ΔyneA::loxP amyE::Phy-yneAΔ15 (EAM55) spotted on the indicated media. (C) Spot titer assay using B. subtilis strains WT (PY79), ΔddcP ΔyneA::loxP amyE::Phy-yneA (EAM48), ΔddcP ΔyneA::loxP amyE::Phy-yneAΔ5 (EAM83), ΔddcP ΔyneA::loxP amyE::Phy-yneAΔ10 (EAM84) and ΔddcP ΔyneA::loxP amyE::Phy-yneAΔ15 (EAM85) spotted on the indicated media. (D) Western blot using antisera against YneA (upper panel) or DnaN (lower panel) using B. subtilis strains WT (PY79), ΔyneA::loxP amyE::Phy-yneA (EAM46), ΔyneA::loxP amyE::Phy-yneAΔ5 (EAM53), ΔyneA::loxP amyE::Phy-yneAΔ10 (EAM54) and ΔyneA::loxP amyE::Phy-yneAΔ15 (EAM55) after growing in the presence of IPTG until an OD600 = 1. (E) Western blot using antisera against YneA (upper panel) or DnaN (lower panel) using B. subtilis strains WT (PY79), ΔddcP ΔyneA::loxP amyE::Phy-yneA (EAM48), ΔddcP ΔyneA::loxP amyE::Phy-yneAΔ5 (EAM83), ΔddcP ΔyneA::loxP amyE::Phy-yneAΔ10 (EAM84) and ΔddcP ΔyneA::loxP amyE::Phy-yneAΔ15 (EAM85) after growing in the presence of IPTG until an OD600 = 1.