Project description:Expression profile of Mycobacterium tuberculosis H37Rv biofilm as induced by DTT (Reduced) 6mM DTT reduced at 6 mM concentration was added to log phase culture of Mtb H37Rv. After 29 hours RNA was isolated and hybridization was done on microarrays
Project description:The global protein expression of Mycobacterium tuberculosis H37Rv, responding to VC treatment (5 mM for 24 h), was monitored via tandem mass tag (TMT)-based quantitative proteomic analysis.
Project description:Mycobacterium tuberculosis (M. tb), the cause of tuberculosis (TB), utilizes the blood circulation to spread systemically and establish infection, and the risk of developing active TB (pulmonary and extrapulmonary) is significantly increased in individuals infected with human immunodeficiency virus (HIV). In this work, we have used DNA microarray analysis to investigate the transcriptome of M. tb replicating in human whole blood from both HIV-negative and HIV-positive donors compared to M. tb grown in Middlebrook 7H9 broth media in order to identify M. tb adaptations to this host environment as well as M. tb mechanisms/factors contributing to increased active and disseminated TB during M. tb/HIV co-infection. We compared the global gene expression of M. tb H37Rv replicating in whole blood from 6 HIV- and 6 HIV+ individulas at 96 hr to M. tb grown to log phase in Middlebrook 7H9 media.
Project description:Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (MTB), which resulted millions of deaths worldwide every year. Although the type strain M. tuberculosis H37Rv was sequenced in 1998, annotation errors of encoding genes have emerged in an endless stream. The annotation errors are particularly severe in the N-terminal and start sites of the genes. In this study, we applied a dimethylation labeling combined with negative enrichment strategy on M. tuberculosis H37Rv to characterize the N-terminal start sites of its annotated encoding genes. Totally, 18,285 peptides and 1,641 N-terminal peptides were identified from all the 2,728 database annotated proteins. The negative N-terminal enrichment strategy allowed the re-annotation of 12 genes’ N-terminal and identification of 6 novel genes in H37Rv. The comparative genomics approach could provide possible help for the re-annotation of 403 mis-annotated genes from Mycobacteriaceae. In addition, we verified the novel gene Rv3409, which was identified with 3 high-confident peptides, with the synthetic peptides and the expression of recombinant protein Rv3409. Altogether, our study and findings contributed to a better understanding of the N-terminal annotation of H37Rv, which will contribute to further biological studies of other species of Mycobacteriaceae in the future.
Project description:Transcriptional profiling of Mycobacterium tuberculosis H37Rv strains comparing control DMSO treated strains with Linezolid treated strains. Goal was to determine the effects of Linezolid against Mycobacterium tuberculosis H37Rv strains.
Project description:Transcriptional profiling of Mycobacterium tuberculosis H37Rv strains comparing control DMSO treated strains with Lupulone treated strains. Goal was to determine the effects of Lupulone against Mycobacterium tuberculosis H37Rv strains.
Project description:Differentially regulated miRNA candidates in H37Rv infected THP-1 cells were analysed with respect to uninfected THP-1 reference samples. THP-1 cells are monocytes differentiated to macrophages after treatment with PMA for 48 hrs. Total RNA was isolated from infected THP-1 cells after 24 hrs of infection, cDNA was synthesized for TLDA real time PCR reaction using TaqMan MicroRNA Reverse Transcription kit and Megaplex Human Pool A and Pool B stem loop RT primers (version 3.0) as per manufacturer’s protocol. Further real time reaction was performed on QuantStudio 12K Flex Real-Time PCR System (Applied Biosystems) by using cDNA (without pre-amplification) on TLDA card A and card B (version 3.0).
