Table 2 Description of the plasmids and strains used in this study
Strain Name | Description | Source |
|---|---|---|
E. coli XL1 Blue | recA1 endA1 gyrA96 thi-1 hsdR17 supE44 relA1 lac [F’ proAB lacIq ZΔM15 Tn10 (Tetr)]. | Stratagene, USA |
E. coli HB101 | F-, thi-1, hsdS20 (rB- mB), supE44, recA13, ara-14, leuB6, proA2, lacY1, galK2, rpsL20 (strr), xyl-5, mtl-1. | Promega, USA |
E. coli BL-21 (λDE3, pLysE) | F–, ompT, hsdSB (rB–, mB–), dcm, gal, λ(DE3), pLysE, Cmr | Promega, USA |
M. tuberculosis H37Rv | Virulent strain of M. tuberculosis. | ATCC27294 |
M. smegmatis mc2155 | Non-pathogenic fast growing mycobacteria | A kind gift from Prof. Anil K Tyagi |
ΔvapC13 | Rv1838c mutant strain of M. tuberculosis. | This work |
ΔvapBC13 | Rv1838c-Rv1839c mutant strain of M. tuberculosis. | This work |
ΔvapC26 | Rv0582 mutant strain of M. tuberculosis. | This work |
ΔvapBC26 | Rv0581-Rv0582 mutant strain of M. tuberculosis. | This work |
ΔvapC13-CT | ΔvapC13 strain of M. tuberculosis complemented with Rv1838c. | This work |
ΔvapBC13-CT | ΔvapBC13 strain of M. tuberculosis complemented with Rv1838c-Rv1839c. | This work |
ΔvapBC13::vapC13OE | ΔvapBC13 strain of M. tuberculosis harbouring anhydrotetracycline-inducible tagged vector, pTetR-int-vapC13. | This work |
ΔvapC26-CT | ΔvapC26 strain of M. tuberculosis complemented with Rv0582. | This work |
ΔvapBC26-CT | ΔvapBC26 strain of M. tuberculosis complemented with Rv0581-Rv0582. | This work |
ΔvapBC26::vapC26OE | ΔvapBC26 strain of M. tuberculosis harbouring anhydrotetracycline-inducible tagged vector, pTetR-int-vapC26. | This work |
H37Rv, pTetR | M. tuberculosis harbouring anhydrotetracycline-inducible vector, pTetR. | |
H37Rv, pTetR-vapC13 | M. tuberculosis harbouring anhydrotetracycline-inducible vector, pTetR-vapC13. | |
H37Rv, pTetR-vapC26 | M. tuberculosis harbouring anhydrotetracycline-inducible vector, pTetR-vapC26. | |
mc2155, pTetR- myc | M. smegmatis strain harbouring anhydrotetracycline-inducible myc tagged vector, pTetR-myc. | This work |
mc2155, pTetR-vapC13 myc | M. smegmatis strain harbouring anhydrotetracycline-inducible myc tagged vector, pTetR-vapC13 myc. | This work |
mc2155, pTetR-vapC26 myc | M. smegmatis strain harbouring anhydrotetracycline-inducible myc tagged vector, pTetR-vapC26 myc. | This work |
Plasmid name | Description | Source |
pGEM-T easy | T/A cloning vector, ampr | Promega, UK |
pYUB854 | Cloning vector, hygr | |
pYUB854ΔvapC13::hygr | pYUB854 with Rv1838c upstream and downstream region flanking the hygromycin resistance gene. | This work |
pYUB854ΔvapBC13::hygr | pYUB854 with Rv1839c upstream and Rv1838c downstream region flanking the hygromycin resistance gene. | This work |
pYUB854ΔvapC26::hygr | pYUB854 with Rv0582 upstream and downstream region flanking the hygromycin resistance gene. | This work |
pYUB854ΔvapBC26::hygr | pYUB854 with Rv0581 upstream and Rv0582 downstream region flanking the hygromycin resistance gene. | This work |
pMAL-c2x | Prokaryotic expression vector with MBP tag at amino-terminus. | New England Biolabs, US |
pMAL-c2x-vapC13 | pMAL-c2x harbouring Rv1838c. | This work |
pLimo | Prokaryotic expression vector with MBP tag at amino-terminus. | |
pLimo-vapC26 | pLimo harbouring Rv0582. | This work |
pTetR | anhydrotetracycline inducible vector system. | |
pTetR-myc | anhydrotetracycline inducible vector system with myc tag at C-terminal. | This work |
pTetR-int | anhydrotetracycline inducible vector system. | |
pTetR-vapC13 | pTetR vector harbouring Rv1838c. | |
pTetR-vapC13 myc | pTetR-myc vector harbouring Rv1838c. | This work |
pTetR-int-vapC13 | pTetR-int vector harbouring Rv1838c. | This work |
pTetR-vapC26 | pTetR vector harbouring Rv0582. | |
pTetR-vapC26 myc | pTetR-myc vector harbouring Rv0582. | This work |
pTetR-int-vapC26 | pTetR-int vector harbouring Rv0582. | This work |
pJEB402 | Constitutive mycobacterial expression integrative vector. | A kind gift from Dr Amit Kumar Pandey |
pJEB402-vapBC13 | pJEB402 harbouring Rv1838c-Rv1839c. | This work |
pJEB402-vapBC26 | pJEB402 harbouring Rv0581-Rv0582. | This work |
pYUB159 | temperature sensitive mycobacteriophage | |
pYUB159ΔvapC13::hygr | pYUB159 derivative to replace Rv1838c with hygromycin resistance gene in M. tuberculosis. | This work |
pYUB159ΔvapBC13::hygr | pYUB159 derivative to replace Rv1838c-Rv1839c with hygromycin resistance gene in M. tuberculosis. | This work |
pYUB159ΔvapC26::hygr | pYUB159 derivative to replace Rv0582 with hygromycin resistance gene in M. tuberculosis. | This work |
pYUB159ΔvapBC26::hygr | pYUB159 derivative to replace Rv0581-Rv0582 with hygromycin resistance gene in M. tuberculosis. | This work |
