Transcriptome analysis of VapC36 overexpression in Mycobacterium tuberculosis
Using Total RNA sequencing, We report the effects of VapC36 overexpression on the transcriptome of M. tuberculosis. For RNA-seq experiments, early-log phase cultures (OD600nm ~ 0.2-0.3) of M. tuberculosis strains harboring either pTetR or pTetR-vapC36  were induced with the addition of 50 ng/ml Atc for 24 h. For total RNA isolation, induced cultures were harvested by centrifugation, washed twice with 1x PBS and lysed by bead beating in Trizol. Total RNA was extracted by phenol-chloroform, precipitated with isopropanol and eluted using Qiagen RNA isolation kit as per manufacturer's instructions. We report that VapC36 resulted in increased transcripts levels of proteins encoding for ribosomal proteins  and copper responsive proteins of M. tuberculosis. Overall design: Cells harbouring pTetR (Vector only) or pTetR VapC36 were used for RNA-Seq analysis. Experiments were performed thrice for each strain.