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KMID : 0880220160540080565
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Journal of Microbiology
2016 Volume.54 No. 8 p.565 ~ p.572
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Mycobacterium tuberculosis gene expression at different stages of hypoxia-induced dormancy and upon resuscitation
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Iona Elisabetta
Pardini Manuela
Mustazzolu Alessandro
Piccaro Giovanni
Nisini Roberto
Fattorini Lanfranco
Giannoni Federico
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Abstract
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The physiology of dormant Mycobacterium tuberculosis was studied in detail by examining the gene expression of 51 genes using quantitative Reverse-Transcription Polymerase Chain Reaction. A forty-day period of dormancy in the Wayne culture model depicted four major transcription patterns. Some sigma factors and many metabolic genes were constant, whereas genes belonging to the dormancy regulon were activated on day 9. In particular, alpha-crystallin mRNA showed more than a 1,000-fold increase compared to replicating bacilli. Genes belonging to the enduring hypoxic response were up-regulated at day 16, notably, transcription factors sigma B and E. Early genes typical of log-phase bacilli, esat-6 and fbpB, were uniformly down-regulated during dormancy. Late stages of dormancy showed a drop in gene expression likely due to a lack of substrates in anaerobic respiration as demonstrated by the transcriptional activation observed following nitrates addition. Among genes involved in nitrate metabolism, narG was strongly up-regulated by nitrates addition. Dormant bacilli responded very rapidly when exposed to oxygen and fresh medium, showing a transcriptional activation of many genes, including resuscitation-promoting factors, within one hour. Our observations extend the current knowledge on dormant M. tuberculosis gene expression and its response to nutrients and to aerobic and anaerobic respiration.
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KEYWORD
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Mycobacterium tuberculosis, H37Rv, dormancy, gene expression
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