TABLE 1.
List of primers used in this study
| Primer name | Sequence 5′–3′ | Use |
|---|---|---|
| Mtb Fwd ctpC::hyg hygN-out2 | 5′-ACCAGTGCTCGCCGAACAGCTTGA-3′ | Amplification of the N- terminal regions of Mtb ctpC |
| Mtb Rvs ctpC::hyg CatC-out2 | 5′-TGCACGGGACCAACACCTTCGTGG-3′ | Amplification of the C-terminal regions of Mtb ctpC |
| prEL8 | 5′-TCGTTAACGACCCGGAAGAAAGGTACAG-3′ | Amplification of the M. tuberculosis rv3269-ctpC operon |
| prEL9 | 5′-ATGGTACCCGAACGGCTGGTGGATTGAC-3′ | Amplification of the M. tuberculosis rv3269-ctpC operon |
| Mtb ctpC Fwd | 5′-ACCCTGGAAGTGGTATCGGACGCG-3′ | Clone Mtb ctpC in pBAD |
| Mtb ctpC Rvs | 5′-GCGGTCCAGGCGGTAGCGGAT-3′ | Clone Mtb ctpC in pBAD |
| Mtb ctpC ANASS Fwd | 5′-GCGGCGATCCTGGCTAACGCGTCGTCG-3′ | Mutation of the TM-MBD (TM8) of Mtb ctpC |
| Mtb ctpC ANASS Rvs | 5′-CGACGACGCGTTAGCCAGGATCGCCGC-3′ | Mutation of the TM-MBD (TM8) of Mtb ctpC |
| Mtb ctpC HNAAA Fwd | 5′-GCGATCCTGCACAACGCGGCTGCTGTGGCGGTGGTG-3′ | Mutation of the TM-MBD (TM8) of Mtb ctpC |
| Mtb ctpC HNAAA Fwd | 5′-CACCACCGCCACAGCAGCCGCGTTGTGCAGGATCGC-3′ | Mutation of the TM-MBD (TM8) of Mtb ctpC |
| Mtb qFwd sigA | 5′-CTCGGTTCGCGCCTACCTCA-3′ | M. tuberculosis Housekeeping gene for qPCR analysis |
| Mtb qRvs sigA | 5′-GCGCTCGCTAAGCTCGGTCA-3′ | M. tuberculosis Housekeeping gene for qPCR analysis |
| Mtb qFwd ctpC | 5′-TCGACGCCATCGTGTTCG-3′ | M. tuberculosis ctpC gene for qPCR analysis |
| Mtb qRvs ctpC | 5′-CTCGGGCTCCCAATCTTTATGC-3′ | M. tuberculosis ctpC gene for qPCR analysis |
| Smeg6058-F | 5′-GTGGCTGATCTGACGGTCGTATCCGACGCG GCCGGCCGCATGCGCGTACACGGCCGCTCTAGAACTAGTGGA-3′ | Hyg cassette amplification with 50 bp of 5′-flanking region locus MSMEG_6058 added to the 5′-end |
| Smeg6058-R | 5′-CTAGTCGATTTCGTACCGGATCAGCCGGGCG CTGTTGGCCACCACCGCAACATGCCTGCAGGTCGACTCT-3′ | Hyg cassette amplification with 50 bp of 3′-flanking region locus MSMEG_6058 added to the 3′-end |
| MSMEG_6058-F | 5′-CATAGGTGAGGAAGTGCCGCCGCCCGCCGC GGGCGACGCCGGCCACGACCACGACCACTGAGTGACCACTGAGTACCGTGGCTGATCTGACGGT CGT-3′ | Hyg cassette amplification with 125 bp of 5′-flanking region locus MSMEG_6058 added to the 5′-end |
| MSMEG_6058-R | 5′-GGCGGCCGACATGGTGAACGCGCCGGTGGG CGGTACGACGACGTGTTGCACACCGGCAGCGTTCGCCGAACGCCCTAGTCGATTTCGTACCGGA-3′ | Hyg cassette amplification with 125 bp of 3′-flanking region locus MSMEG_6058 added to the 3′-end |
| intSmeg6058-For | 5′-TTGATTCTCTTGGGGATTCG-3′ | Deletion MSMEG_6058 verification |
| intSmeg6058-Rev | 5′-ATGATGCGACCTATGGTGGT-3′ | Deletion MSMEG_6058 verification |
| Smeg6058-V1 | 5′-GTGATGGCAGAGGCGCGCGAA-3′ | Deletion MSMEG_6058 verification |
| Smeg6058-V2 | 5′-GTCAAGCGAATCGGCCGCCAC-3′ | Deletion MSMEG_6058 verification |
| Fwd EcoRI ctpc | 5′-ACTGGAATTCGTGGCTGATCTGACGGTCGT ATC-3′ | Clone SMEG_6058 in pJEB402 |
| Rvs HpaI ctpc | 5′-ACTGGTTAACCTAGTCGATTTCGTACCGG-3′ | Clone SMEG_6058 in pJEB402 |
| ctpC_smeg fwd | 5′-GTGGCTGATCTGACGGTCGTATC-3′ | Clone SMEG_6058 in pBAD |
| ctpC_smeg rvs | 5′-CTAGTCGATTTCGTACCGGATCAG-3′ | Clone SMEG_6058 in pBAD |
| Msm_2758 qFwd sigA | 5′-GAAGACACCGACCTGGAACT-3′ | M. smegmatis Housekeeping gene for qPCR analysis |
| Msm_2758 qRvs sigA | 5′-GACTCTTCCTCGTCCCACAC-3′ | M. smegmatis Housekeeping gene for qPCR analysis |
| Msm qFwd ctpC | 5′-CGTGGTGGCCCTTACGGTTC-3′ | M. smegmatis ctpC gene for qPCR analysis |
| Msm qRvs ctpC | 5′-CGATCGCAACCTGCTCGTGT-3′ | M. smegmatis ctpC gene for qPCR analysis |