tuberculosis (data not shown). Overexpression of Mce2R reduces M. tuberculosis replication in a mouse model of infection In order to examine the infection and survival pattern Selleckchem GSK690693 of the MtΔmce2R mutant in vivo, we used the intratracheal route to infect BALB/c mice [8], and determined lung colonization by counting bacterial colony forming units (CFUs). At 26 and 35 days post-infection, the number of CFUs in lungs of animals inoculated with the MtΔmce2R mutant was equivalent than that of the animals inoculated with the parental strain (Figure 2). However, the introduction of a constitutively expressed mce2R gene into the

MtΔmce2R mutant (MtΔmce2RComp) significantly reduced the replication of M. tuberculosis in lungs at 26 and 35 days post-infection (p < 0.05). This result led us to hypothesize that the expression of the mce2 operon was over-repressed in the complemented strain due to the overexpression of Mce2R. To test this possibility, we assessed the in vitro expression of mce2R and yrbE2A in the complemented and the wild type strains at both the early and late exponential phases of bacterial growth. The level of transcription of mce2R in the complemented strain was higher than in the wild type strain (p < 0.05) at the exponential and stationary growth phases (Table 1). At the early exponential phase,

the differences in the amount of yrbE2A mRNA between both strains were not statistically significant whereas at the late exponential phase there was a significant reduction in yrbE2A mRNA (p < 0.05) PF-6463922 in vivo in the complemented strain as compared with that in the wild type strain

(Table 1). Figure 2 Replication of the MtΔmce2R mutant, the wild type and complemented strains in mouse lungs after intratracheal inoculation. Groups of mice were GS-9973 clinical trial infected by intratracheal injection of wild type (white bars), MtΔmce2R (black bars), MtΔmce2RComp (grey bars). At 1, 26 and 35 days post-infection, mice were sacrificed and viable bacteria present in the lungs were recovered. The results are expressed as the mean number of CFUs ± standard deviations in five mice. These data are based on one of two independent experiments with similar results. *(p < 0.05) significantly different from values of the wild Nintedanib (BIBF 1120) type strain. The lack of Mce2R only affects the expression of mce2 operon during the in vitro culture of M. tuberculosis To define the Mce2R regulon we performed a whole-genome in vitro expression profiling on the mutant and wild-type parental H37Rv strains. The analysis of gene expression data showed that about 99.6% of all genes showed fold changes equal or greater than 1.2 (absolute value) (Additional file 1: Table S1), indicating that most of the genes were similarly expressed in the mutant and the wild type strains. We found only 16 genes that were overexpressed in MtΔmce2R with fold changes >1.2.