Journal: Future Microbiology
Article Title: LdtMav2, a nonclassical transpeptidase and susceptibility of Mycobacterium avium to carbapenems
Figure Lengend Snippet: L,D-transpeptidases in mycobacteria and expression of Ldt Mav2 . (A) Cladogram of L,D-transpeptidases from various mycobacteria. Multiple alignment and phylogeny analysis of L,D-transpeptidases from Mycobacterium tuberculosis (O), Mycobacterium smegmatis (□), Mycobacterium absessus (Δ), Mycobacterium leprae (∇), Mycobacterium marinum (◊) and Mycobacterium avium (□). Ldt Mav2 is highlighted as a red-closed square. (B) Western blot analysis of cell wall and cytosolic fractions of M. avium harvested at logarithmic (log) and stationary (stat) phases of growth and probed with antibody against Ldt Mav2 . His6-tagged Ldt Mav2 is included as a control (far right). (C) Levels of Ldt Mav2 in the presence of 100 µM iron (lane 1), 40 µM iron (lane 2), 0.05% SDS (lane 3), 2 µg/ml crystal violet (lane 4), 1 µg/ml tebipenem (lane 5), H 2 O (lane 6), Middlebrook 7H9-enriched medium (lane 7) and His6-tagged Ldt Mav2 as a control (lane 8).
Article Snippet: • Bacterial strains & growth conditions M. avium 104, a clinical isolate [ ], was used in this study and was grown in Middlebrook 7H9 broth (Becton and Dickinson, Sparks, MD, USA) supplemented with 10% (v/v) oleic acid/albumin/dextrose/catalase (OADC), 0.2% glycerol, 0.05% Tween-80 and 50 µg/ml cycloheximide (7H9 complete medium) at 37°C with constant shaking.
Techniques: Expressing, Western Blot