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Volume 154, Issue 8

Characterization of genes differentially expressed within macrophages by virulent and attenuated identifies candidate genes involved in intracellular growth Free

Abstract

To identify genes involved in the intracellular survival of we compared the transcriptomes of virulent (H37Rv) and attenuated (H37Ra) strains during their interaction with murine bone-marrow-derived macrophages. Expression profiling was accomplished via the bacterial artificial chromosome fingerprint array (BACFA) technique. Genes identified with BACFA, and confirmed via qPCR to be upregulated in the attenuated H37Ra at 168 h post-infection, were , and . Genes upregulated in the virulent H37Rv were , and . Further qPCR analysis of these genes at 4 and 96 h post-infection revealed that the operon (encoding the fumarate reductase enzyme complex) is expressed at higher levels in the virulent H37Rv at earlier time points while the expression of and is higher in the virulent strain throughout the course of infection. Assessment of transcripts in oxygen-limited cultures of H37Ra and H37Rv showed that the attenuated strain displayed a lag in and expression at the onset of microaerophilic culture, when compared to microaerophilic cultures of H37Rv and aerated cultures of H37Ra. Lastly, treatment of intracellular bacteria with a putative inhibitor of fumarate reductase resulted in a significant reduction of bacterial growth.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): BACFA, bacterial artificial chromosome fingerprint array , BM-MΦ, bone-marrow-derived macrophages , FRD, fumarate reductase , MNPO, mercaptopyridine-N-oxide , qPCR, quantitative real-time PCR , SDH, succinate dehydrogenase and TB, tuberculosis
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2008-08-01
2024-04-19
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Characterization of genes differentially expressed within macrophages by virulent and attenuated Mycobacterium tuberculosis identifies candidate genes involved in intracellular growth
Microbiology 154, 2291 (2008); https://doi.org/10.1099/mic.0.2008/019661-0
/content/journal/micro/10.1099/mic.0.2008/019661-0
/content/journal/micro/10.1099/mic.0.2008/019661-0
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