Fluorescent M. tuberculosis Strains for Infection Studies

Mycobacterium tuberculosis Erdman (American Type Culture Collection 35801) was the parental strain used for all experiments. The fluorescent reporters smyc′::mCherry, hspx′::GFP/smyc′::mCherry, and hsp60′::GFP were previously described (Abramovitch et al., 2011 (link); Dhandayuthapani et al., 1995 (link); Sukumar et al., 2014 (link); Tan et al., 2013 (link)). Bacteria were grown at 37°C to mid-log phase in MiddleBrook 7H9 broth supplemented with 10% oleic acid/albumin/dextrose/catalase (OADC Enrichment; Becton, Dickinson and Company), 0.2% glycerol, and 0.05% tyloxapol (Sigma-Aldrich). BCG (Pasteur) was grown as described above. Hygromycin B (50 mg/ml) was used as a selection marker for the fluorescent strains. For mice infection, aliquots were frozen in 10% glycerol, titered, and stored at −80°C until use.