TnSeq of M. tuberculosis H37Rv in vaccinated mice
Use of using TnSeq (sequencing of Himar1 transposon insertion mutant libraries) to investigate differences in conditionally essential genes in M. tuberculosis H37Rv in infections of mice vaccinated by different strategies. This data was collected collected at Harvard School of Public Health and analyzed in the lab of Dr. Allison Carey, Department of Pathology The University of Utah. The TnSeq library was generated with HImar1 in M. tuberculosis H37Rv following protocols in (Long, 2015, PMID: 25636614). C57BL/6 mice were vaccinated by one of several routes, and then challenged with the H37Rv Tn library. Samples were harvested from spleens at 14 or 28 days post-infection and outgrown on plates with 7H10 medium. Vaccination types included: 'BCG-SQ' (vaccination of mice with M. bovis BCG strain subcutaneously), 'BCG-IV' (vaccination of mice with M. bovis BCG strain by tail-vein injection), 'naive' (no vaccination prior to Tn library infection), 'aerosol' (prior infection with H37Rv and cured with antibiotics), and 'lprG' (vaccinated with a null mutant of H37Rv with knockout of lprG). DNA samples were extracted and prepared for sequencing by PCR-amplification (see Long, 2015). Each sample was sequenced on two Illumina runs, producing150x150 bp PE reads containing transposon:genomic junctions.