wild type bacterium  (DSMZ)

Journal: Microbiology Spectrum

Article Title: Characterization of DNA Processing Protein A (DprA) of the Radiation-Resistant Bacterium Deinococcus radiodurans

doi: 10.1128/spectrum.03470-22

Figure Lengend Snippet: Domain architecture of DprA. (A) Domain architecture of DprA proteins of select bacteria ( S. pneumoniae , R. palustris , D. radiodurans , and H. pylori ). The central Rossmann fold (RF) is conserved and present in all DprA proteins, while the N-terminal SAM domain is common in all DprA proteins except that of H. pylori . An additional C-terminal DML1 domain is present in the DprA proteins of D. radiodurans and R. palustris . (B) Multiple-sequence alignment of DprA proteins. Red labels represent conserved putative amino acids responsible for the DprA-RecA interaction (I287, G253, and D277), the DprA-DprA interaction (L293, and H284), and the interaction with DNA (R137, R234, and F229), as predicted by in silico analysis.

Article Snippet: The wild-type bacterium D. radiodurans R1 (ATCC 13939) and its mutants were grown in tryptone-glucose-yeast extract (TGY) medium (1% Bacto tryptone, 0.1% glucose, 0.5% yeast extract) with the appropriate antibiotic, as described previously ( ).

Techniques: Bacteria, Sequencing, In Silico

Journal: Microbiology Spectrum

Article Title: Characterization of DNA Processing Protein A (DprA) of the Radiation-Resistant Bacterium Deinococcus radiodurans

doi: 10.1128/spectrum.03470-22

Figure Lengend Snippet: DrRecA-DrDprA interaction in surrogate E. coli cells ( ex vivo ), in D. radiodurans cells ( in vivo ), and in vitro . (A) A total of 2.5 μM DrDprA protein was immobilized on a gold sensor chip, followed by incubation with different concentrations (0.5 to 3 μM) of DrRecA. Surface plasmon resonance (SPR) signals were recorded, and data were processed as described in Materials and Methods. The data given are representative of data from experiments repeated three times. RU, response units. (B) T18 and T25 tags of adenylate cyclase alone and with the dprA and recA genes of D. radiodurans were cloned into BACTH plasmids. These plasmids were transformed into the E. coli BTH101 host. The interactions of proteins tagged with T18 and T25 were monitored as white-blue colonies. RecA-C18 and RecA-C25 were used as positive controls, while C18 and C25 tag-expressing cells were used as negative controls. (C) Cell extracts of D. radiodurans cells coexpressing C18-DrDprA and His-DrRecA from plasmids pVHSM and pBAD, respectively, used for immunoprecipitation assays. Immunoprecipitation was done using anti-His antibody, and immunoprecipitates were separated on an SDS-PAGE gel, followed by immunoblot detection using antibodies against the T18 domain of CyaA (anti-T18 immunoblot), as detailed in Materials and Methods. M, molecular marker.

Article Snippet: The wild-type bacterium D. radiodurans R1 (ATCC 13939) and its mutants were grown in tryptone-glucose-yeast extract (TGY) medium (1% Bacto tryptone, 0.1% glucose, 0.5% yeast extract) with the appropriate antibiotic, as described previously ( ).

Techniques: Ex Vivo, In Vivo, In Vitro, Incubation, SPR Assay, Clone Assay, Transformation Assay, Expressing, Immunoprecipitation, SDS Page, Western Blot, Marker