Table 3 Plasmids and bacterial strains constructed in the study
Plasmid | Source of description | Reference/origin |
|---|---|---|
E. coli DH5α | F−endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG ϕ80dlacZΔM15 Δ(lacZYA-argF)U169, hsdR17(rK-mK+), λ- | Invitrogen |
E. coli BL21 (DE3) | F−ompT gal dcm lon hsdSB(rB−mB−) λ(DE3 [lacI lacUV5-T7 gene 1 ind1 sam7 nin5] | Invitrogen |
M. smegmatis mc2155 | Lab stock | |
pGEX 4 T-1 | lacTq, 4.9 kb, Ampr, GST gene fusion vector | GE Healthcare |
pRSET-B | lacTq, 2.8 kb, AmpR, 6xHis (N-term) gene fusion vector | Invitrogen |
p0004- Sac B | Suicide recombination delivery vector carrying hygR-SacB for gene disruption, hygR | |
PhAE159 | Conditionally replicating shuttle phasmid vector | William R Jacobs Jr (unpublished) |
pMV261 | E. coli mycobacterial shuttle vector, KanR, hsp60 promoter | Strover et al. (1991) |
Constructs in this study | ||
pGBRJ1 | pRSET-B vector carrying the coding region of Rv0148 gene from M. tuberculosis, AmpR | This study |
pGBRJ2 | pGEX 4 T-1 vector carrying the coding region of Rv0148 gene from M. tuberculosis, AmpR | This study |
pGBRJ3 | pRSET-B vector carrying the coding region of Htdy gene from M. tuberculosis, AmpR | This study |
Δ0148 | The deletion of Rv0148 gene from M. tuberculosis | This study |
CΔ0148 | pMV261 vector carrying the coding region of Rv0148 gene from M. tuberculosis, KanR expressed in M. tuberculosis (complementation) | This study |