TABLE 1.
Strains, plasmids, and primers used in this study
| Primer, plasmid, or strain | Relevant detailsa | Source |
|---|---|---|
| Primers | ||
| 43pr1 | 5′-TAGCGGCCGCACTGAAGCAT-3′ | This study |
| 43pr2 | 5′-GACTCTAGACGTGTCTAAGAG-3′ | This study |
| sigA1 | 5′-GGTACCGTCGCAGATACGACGCAC-3′ | This study |
| sigA2 | 5′-TCTAGAATACACCCCTTCGGTCGT-3′ | This study |
| 16s1 | 5′-GGTACCTCGGTGCCGAGATCGAAC-3′ | This study |
| 16s2 | 5′-TCTAGACCCAAACACTCCCTTTGG-3′ | This study |
| 60P1 | 5′-TAGCGGCCGCTAGAGGTGAC-3′ | This study |
| 60P2 | 5′-GCATCTAGATGCGAAGTGATTC-3′ | This study |
| Fluc1 | 5′-TCTAGATGGAAGACGCCAAAAACAT-3′ | This study |
| Fluc2 | 5′-GGAATTCTTACACGGCGATCTTTC-3′ | This study |
| Rluc1 | 5′-TCTAGAATGACTTCGAAAG-3′ | This study |
| Rluc2 | 5′-GAATTCTTATTGTTCATTTTTGAG-3′ | This study |
| sigART1 | 5′-GTGGCAGCGACCAAAGCAAG-3′ | This study |
| sigART2 | 5′-CACTAGCGGACTTCGCCGCT-3′ | This study |
| 60RT1 | 5′-CCAAGACAATTGCGTACGACGA-3′ | This study |
| 60RT2 | 5′-CCACCCCACTTCTTTTCCAGGA-3′ | This study |
| 16sRT1 | 5′-CCTGGCTCAGGACGAACGCT-3′ | This study |
| 16sRT2 | 5′-TCCCGAAGTGCAGGGCAGAT-3′ | This study |
| FlucRT1 | 5′-CCATTCTATCCGCCTGGAAG-3′ | This study |
| FlucRT2 | 5′-CGTAAGTGATGTCCACCTCG-3′ | This study |
| Plasmids | ||
| pGL3-Basic | Plasmid vector with Fluc ORF, Ampr | Promega |
| pFN11A | Plasmid vector with Rluc ORF, Ampr | Promega |
| pTZhsp60p | Plasmid vector carrying hsp60p, Ampr | This study |
| pTZsigAp | Plasmid vector carrying sigAp, Ampr | This study |
| pTZ16Sp | Plasmid vector carrying 16S rRNAp, Ampr | This study |
| pTZFluc | Plasmid vector carrying Fluc ORF, Ampr | This study |
| pTZRluc | Plasmid vector carrying Rluc ORF, Ampr | This study |
| pTZ43p | Plasmid vector carrying 43p, Ampr | This study |
| 306h:Fluc | Integrative vector carrying Fluc ORF, Hygr | This study |
| 306h:hsp60pr-Fluc | Integrative vector carrying hsp60p and Fluc ORF, Hygr | This study |
| 306h:sigApr-Fluc | Integrative vector carrying sigAp and Fluc ORF, Hygr | This study |
| 306h:16S p-Fluc | Integrative vector carrying 16S rRNAp and Fluc ORF, Hygr | This study |
| 206:43p-Rluc | Extrachromosomal plasmid vector carrying 43p and Rluc ORF | This study |
| Bacterial strains | ||
| NEB10 | Δ(lacZ)M15 mcrA galU recA1 endA1 nupG rpsL (Strr) | NEB |
| M. bovis BCG | Pasteur strain | Lab stock |
| BCG hsp60p-Fluc | M. bovis BCG carrying 306h:hsp60p-Fluc, Hygr | This study |
| BCG sigAp-Fluc | M. bovis BCG carrying 306h:sigAp-Fluc, Hygr | This study |
| BCG 16S rRNAp-Fluc | M. bovis BCG carrying 306h:16S-p-Fluc, Hygr | This study |
| BCG hsp60p-Fluc 43p-Rluc | M. bovis BCG carrying 306h:hsp60p-Fluc:206-43p-Rluc, Hygr Kmr | This study |
a
Ampr, ampicillin resistant; Hygr, hygromycin resistant; Kmr, kanamycin resistant.