GENES DIFFERENTIALLY EXPRESSED BY Mycobacterium tuberculosis AFTER EXPOSURE TO RUTHENIUM PHOSPHINIC COMPOUND AND ISONIAZID

Abstract

Background- The evaluation of the effects of new compounds and nonconventional anti-tuberculous drugs have grown and become increasingly more popular in recent years. Studies have shown anti-tuberculous activity for Ruthenium complexes, including organometallic compounds containing phosphine ligands such as picolinic acid generating great expectations and hopes.Methods- The Representational Difference Analysis (RDA) was applied in order to gain insight about differences in expression of Mycobacterium tuberculosis H37Rv exposed to [Ru(dppb)(pic)(bypy)]PF6 (SCAR1) and isoniazid (INH). Total RNA was extracted from the bacillus not exposed and exposed to SCAR1 and INH separately at concentration of MIC for 12 hours at 35°C. RDA was carried out and differentially expressed products were sequenced.Results- RDA-sequencing identified, for both compounds, orthologs that encode hypothetical and predict proteins. One related cell wall synthesis gene, identified by RDA, and genes related to INH target as inhA, katG and ahpC had their expression confirmed and quantified by real-time PCR. The gene encoding the cell wall associated hydrolase was induced 4.627 and 1.189, inhA 0.983 and 1.027, katG 1.111 and 1.345 and ahpC 1.063 and 1.039 fold after exposure to SCAR1 and INH respectively, compared to not exposed growth.Conclusion- The RDA brings, for the first time, directions to study related genes with metabolic pathways of SCAR1. RDA and Real-Time PCR highlight the idea that one of the SCAR1 interaction, in M tuberculosis may be in the cell wall biosynthesis considering the differential expression of a cell wall hydrolase and warrants further investigation

Anti- Mycobacterium tuberculosis activity of platinum(II)/ N , N -disubstituted- N ′-acyl thiourea complexes

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Next-generation sequencing of representational difference analysis products for identification of genes involved in diosgenin biosynthesis in fenugreek (Trigonella foenum-graecum)

“…Representational difference analysis is a powerful and sensitive tool for the identification of differentially expressed genes in two different cDNA populations and it allows for detection of changes in mRNA expression by selective enrichment without any prior knowledge of the gene in question (Leite et al 2013 ). Moreover, cDNA-RDA is capable of detecting both relative (greater than three- to five-fold) and absolute differences in expression (Hubank and Shatz 1999 ).…”

Structural Changes and Differentially Expressed Genes in Pseudomonas aeruginosa Exposed to Meropenem-Ciprofloxacin Combination

“…The literature on this subject has been quite variable (40)(41)(42). A long exposure time and different concentrations may be related to toxic and side effects and may not represent the primary target of the antimicrobial (23). In the present study, we used 60 min of exposure to 0.5ϫ MIC (meropenem, ciprofloxacin, and the MCC), which represents the beginning of the reduction of viable cells, and antimicrobial concentrations with better synergistic action, assessed by time-kill assays.…”