Table 1.
Bacterial strains and plasmids used in this study
| Strain/plasmid | Description | Source/reference |
|---|---|---|
| Strains | ||
| M. tuberculosis H37Rv | Virulent laboratory strain | ATCC* |
| M. tuberculosis H37Ra | Avirulent laboratory strain | CWRU† |
| M. smegmatis mc2155 | ept-1 | AECOM‡ |
| E. coli HB101 | F–, Δ(gpt-proA)62 leuB6 glnV44 ara-14 galK2 lacY1 Δ(mcrC-mrr) rpsL20(Strr) xyl-5 mtl-1 recA13 thi-1 | UCSF§ |
| E. coli JM109 | e14− (McrA−) recA1 endA1 gyrA96 thi-1hsdR17 supE44 relA1 Δ(lac-proAB) | Stratagene |
| E. coli BL21(DE3) | F–, ompT hsdSB ( ) gal dcm (DE3) |
Invitrogen |
| LIX11 | H37Rv ΔpknK | This study |
| LIX16 | LIX11 : : pYA1580 | This study |
| Plasmids | ||
| pGEM-T Easy | AmpR, TA cloning vector for PCR products | Promega |
| pET SUMO | KanR, 6xHis-SUMO expression vector | Invitrogen |
| pYUB854 | HygR, cosmid vector used in construction of the phasmid carrying the allelic-exchange substrate | Bardarov et al. (2002) |
| pMV306 | KanR, mycobacterial integrating vector | Stover et al. (1991) |
| pSH276 | Rv0195 gene in pQE40ΔDHFR | Haydel & Clark-Curtiss (2004) |
| pSH337 | clpC gene in pGEM-T | Haydel & Clark-Curtiss (2004) |
| pSH317 | devR gene in pQE40ΔDHFR | Haydel & Clark-Curtiss (2004) |
| pSH281 | Rv2027c gene in pQE40ΔDHFR | Haydel & Clark-Curtiss (2004) |
| pSH334 | regX3 gene in pZERO/Km | Haydel & Clark-Curtiss (2004) |
| pYA1620 | pknA gene (C-term) in pGEM-T Easy | This study |
| pYA1537 | pknB gene in pGEM-T Easy | This study |
| pYA1538 | pknD gene in pGEM-T Easy | This study |
| pYA1539 | pknE gene in pGEM-T Easy | This study |
| pYA1540 | pknF gene in pGEM-T Easy | This study |
| pYA1541 | pknG gene in pGEM-T Easy | This study |
| pYA1621 | pknH gene (N-term) in pGEM-T Easy | This study |
| pYA1542 | pknI gene in pGEM-T Easy | This study |
| pYA1543 | pknJ gene in pGEM-T Easy | This study |
| pYA1544 | pknK gene (N-term) in pGEM-T Easy | This study |
| pYA1545 | pknK gene (C-term) in pGEM-T Easy | This study |
| pYA1546 | pknL gene in pGEM-T Easy | This study |
| pYA1556 | Full-length pknK gene in pET SUMO | This study |
| pYA1619 | 3′ and 5′ flanking regions of pknK gene cloned in the multiple cloning sites of pYUB854 | This study |
| pYA1580 | pknK gene+300 bp upstream region cloned in XbaI/HindIII sites in pMV306 | This study |
*ATCC no. 25618, American Type Culture Collection.
†ATCC no. 25177, obtained from Dr Richard F. Silver, Case Western Reserve University, OH, USA.
‡Dr William R. Jacobs, Jr, Albert Einstein College of Medicine, NY, USA.
§Dr Herbert Boyer, University of California, San Francisco, CA, USA.