Transcriptome analysis of VapC13 and VapC26 overexpression in Mycobacterium tuberculosis
Using Total RNA sequencing, We report the effects of VapC13 and VapC26 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-vapC13 or pTetR-vapC26 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 VapC13 or VapC26 resulted in increased transcripts levels of stress responsive genes of M. tuberculosis. Overall design: Cells harbouring pTetR (Vector only) or pTetR VapC13 or pTetR VapC26 were used for RNA-Seq analysis. Experiments were performed thrice for each strain.