Inducible expression of HigB toxin in Mycobacterium tuberculosis results in growth inhibition, reduced abundance of a subset of mRNAs and cleavage of tmRNA
Inducible expression of HigB toxin in Mycobacterium tuberculosis results in growth inhibition, reduced abundance of a subset of mRNAs and cleavage of tmRNA
M. tuberculosis H37Rv (ATCC 25618) was grown at 37C in modified Dubos medium (Difco)  supplemented with 4% Dubos medium albumin (Difco) and 0.5% w/v glycerol. RNA was prepared using a FastRNA pro blue kit (Qbiogene). Contaminating DNA was removed using a TURBO DNA-free kit (Ambion) 2-3 ug of total RNA were fragmented (Ambion Cat # AM8740) and strand-specific cDNA libraries were constructed using the Illumina directional mRNA-Seq protocol (Part # 15018460 Rev. A) but with exclusion of polyA-tail and size selection to capture all RNA species. Briefly, this protocol ligates the Illumina v1.5 small RNA 3prime adapter followed by a 5prime adapter to preserve strand specificity.
Experimental Factor: genotype
empty vector