Table 1.
Strains and plasmids used for the generation of Δglpx and glpx complement strains.
Table 2.
List of primers used for the generation of Δglpx.
Fig 1.
Colony PCR and Southern Blot confirming the deletion of glpX gene a) Colony PCR results of the potential double cross overs (DCOs): Lane 1: 1kb ladder, Lane 2: PCR amplification product of suicidal delivery vector FM152 is about 2kb, Lane 3: PCR product of one such potential DCO (Colony 11) is 2kb, Lane 4: PCR product of a WT Mtb is about 3kb, b) Southern blot: Lane 1: WT genomic DNA digest with BamH1 which gives a fragment of about 610 bp (lowermost band in lane 1 and 3), Lane 2: Genomic DNA digest of a potential DCO (Colony 3) with the 610 bp fragment missing, Lane 3: Same as Lane 1, Lane 4: Genomic DNA digest of a potential DCO (Colony 11) with the 600 bp fragment missing.
Fig 2.
In vitro growth profile of ΔglpX, WT Mtb and glpX complement in a) 7H9 medium + OADC enrichment, b) 7H9 medium with no additional carbon source.
Growth profiles are representative of a triplicate data set.
Fig 3.
In vitro growth profile of ΔglpX, WT Mtb and glpX complement on defined (or individual) carbon source(s).
Growth profile in 7H9 medium a) 0.2% glycerol, b) 0.2% Acetate, and c) 0.2% dextrose. Growth profiles are representative of a triplicate data set.
Fig 4.
In vitro growth profile of ΔglpX, WT Mtb and glpX complement on gluconeogenic carbon source(s).
Growth profile in 7H9 medium a) 0.1% oleic acid, and b) 0.1% valeric acid Growth profiles are representative of a triplicate data set.
Fig 5.
In vitro growth profile of ΔglpX, WT Mtb and glpX complement on combination of carbon sources.
Growth profile in 7H9 medium and a) 0.1% each of glycerol and acetate, b) 0.1% each of dextrose and acetate, and c) 0.1% each of dextrose and glycerol. Growth profiles are representative of a triplicate data set.
Fig 6.
glpX is essential for growth in acute phase and survival during the chronic phase of Mtb infection in mice.
Invivo growth and survival plot for ΔglpX compared to WT Mtb and the glpX complement. Data represents the mean±s.d. of six mice per time point.
Fig 7.
glpX gene deletion does not completely abolish the FBPase activity in Δglpx: FBPase Activity was measured in nmol/min/mg protein in crude extracts, mean of two determinations, limit of detection = 0.4.
All values are with a substrate-free control (no FBP) subtracted. The reported readings are the average of 6 measurements coming from duplicate protein samples (n = 3X2 = 6).
Table 3.
MICs for standard antibacterial drugs as determined by the MABA assay.