Journal: Cell systems
Article Title: Synthetically programmed antioxidant delivery by a domesticated skin commensal.
doi: 10.1016/j.cels.2025.101169
Figure Lengend Snippet: Figure 1. Development of a toolbox for C. acnes genetic engineering (A) List of all tools developed in this study. (B) Schematics of C. acnes replicative plasmid optimizations. (C) Average of population median mCherry fluorescent values of three independent replicates for nine different constitutive endogenous promoters, a synthetic consensus promoter (BBa_J23119) from E. coli, and a control harboring a no-insert plasmid (Empty) for exponential and stationary growth phases. (D) Average of population median fluorescent values for C. acnes harboring a fluorescent reporter (colored bars) compared with a no-insert plasmid control (gray bars). Values depicted above bars indicate average fold change of three independent replicates. (E) Pictures representative of the C. acnes KPA171202 wild-type strain grown either as a lawn or as single colonies in a Brucella media agar plate, supplemented or not with X-gal.
Article Snippet: REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Mouse anti-His antibody Bio-Rad Cat# MCA1396GA; RRID: AB_566361 Horseradish peroxidase-coupled anti-mouse antibody SantaCruz Cat# sc516102; RRID: AB_2687626 Bacterial and virus strains E. coli NZYɑ NZYTech MB01008 E. coli EC-24 Knödlseder et al.31 CECT30749 C. acnes KPA171202 DSMZ DSM 16379 C. acnes 09-9 Davidsson et al.43 RefSeq GCF_002600615.1 C. acnes AD24 Lood et al.64 N/A C. acnes H1 Lab isolate Lab collection N/A C. acnes KPA171202 DLysA This study N/A C. acnes KPA171202 DHisB This study N/A C. acnes KPA171202 DProA This study N/A Chemicals, peptides, and recombinant proteins Brucella Medium Condalab 1012 GasPak EZ Anaerobe Pouch System BD BD260683 AnaeroGen System Thermo Scientific AN0025A Brain-heart infusion (BHI) Medium Condalab 1400 DMEM F-12 medium lacking aminoacids and glucose (see Table S1 for the full components list used for this media) US Biological D9807-02A KAPA HiFi Roche KK2601 Phanta Max Vazyme 001 NuPAGE 4-12% gel Invitrogen NP0322BOX Immobilon-P Transfer Membrane Merck Millipore IPVH00010 Tris buffered saline Bio-Rad 1706435 Pierce ECL Western Blotting Substrate Thermo Fisher Scientific 32106 EpiLife medium Gibco MEPI500CA Human keratinocyte culture supplement (HKGS) Gibco S0015 CnT-Prime CELLnTEC CNT-PR Hanks’ balanced salt solution (HBSS) Capricorn Scientific HBSS-2A 6-chloromethyl-2’,7’-dichlorodihydrofluorescein diacetate (CM-H2DCFDA) Invitrogen C6827 DNaseI Invitrogen 18068015 PowerUp SYBR Green Master Mix Life Technologies A25742 2,4-Diacetylphloroglucinol (DAPG) Cayman Chemical 16345 Anhydrotetracycline hydrochloride (aTc) Sigma 94664-10MG Critical commercial assays QIAquick PCR purification Qiagen 28104 QIAquick gel extraction Qiagen 28704 NZYMiniprep kit NZYTech MB01008 PureLink Maxiprep kit Invitrogen K210017 SOD activity assay kit Sigma CS0009 E.Z.N.A.
Techniques: Plasmid Preparation, Control
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