Table 1.
Bacterial strains, plasmids, and cell line used in this study
| Strains, plasmids, and cell line | Description | Reference or source |
|---|---|---|
| Strains | ||
| E. coli XL-1 Blue | endA1 gyrA96(nalR) thi-1 recA1 relA1 lac glnV44F′[::Tn10 proAB+ lacIq Δ(lacZ)M15] hsdR17(rK− mK+) | Stratagene, Heidelberg, Germany |
| E. coli HB101 | F− (gpt-proA)62 leuB6 glnV44 ara-14 galK2 lacY1 (mcrC-mrr) rpsL20 (Strr) xyl-5 mtl-1 recA13 | Life Technologies, CA |
| M. tuberculosis | M. tuberculosis H37Rv expressing recombineering proteins gp60 and gp61 | This study |
| H37Rv ΔbfrA | M. tuberculosis H37Rv bfrA mutant | This study |
| H37Rv ΔbfrB | M. tuberculosis H37Rv bfrB mutant | This study |
| H37Rv ΔbfrAΔbfrB | M. tuberculosis H37Rv bfrA bfrB double mutant | This study |
| H37Rv ΔbfrA Comp | H37Rv ΔbfrA complemented with wild-type bfrA | This study |
| H37Rv ΔbfrB Comp | H37Rv ΔbfrB complemented with wild-type bfrB | This study |
| Plasmids | ||
| pYUB854 | Cloning vector with hygromycin resistance gene cassette flanked by two multiple cloning sites | 3a |
| pSD7 | Mycobacterial promoter cloning vector carrying kanamycin resistance gene and promoterless chloramphenicol resistance gene | 7a |
| pJV53 | Mycobacterium-E. coli shuttle vector encoding recombineering proteins gp60 and gp61 | 25 |
| pVRΔA | pYUB854 with bfrA::hyg | This study |
| pVRΔB | pYUB854 with bfrB::hyg | This study |
| pVRΔAB | pYUB854 with bfrB::CATtrrn | This study |
| pVR.BfrA | pSD7 carrying bfrA gene with native promoter | This study |
| pVR.BfrB | pSD7 carrying bfrB gene with native promoter | This study |
| Cell line | ||
| THP-1 | Human acute monocytic leukemia cell line | NCCS, Pune, India |