TABLE 1.
B. anthracis strains, plasmids, and oligonucleotides used in this study
| Strain, plasmid, or nucleotide | Relevant genotype or sequence (5′→3′) | Source or reference |
|---|---|---|
| Strains | ||
| 7702 | Sterne strain; pXO1+ | Laboratory stock |
| RBAF140 | 7702 pagA-lacZ transcriptional fusion integrated adjacent to pagA | 53 |
| RBAF143 | 7702 lef-lacZ transcriptional fusion integrated adjacent to lef | 53 |
| RBAF144 | 7702 cya-lacZ transcriptional fusion integrated adjacent to cya | 53 |
| 7702-XFI | 7702 atxA-lacZ transcriptional fusion | J.-C. Sirard, personal communication |
| BATX1 | 7702 ΔatxA | 52 |
| 7CodYS | 7702 codY::spc | This study |
| 140CodYS | RBAF140 codY::spc | This study |
| 143CodYS | RBAF143 codY::spc | This study |
| 144CodYS | RBAF144 codY::spc | This study |
| 7CodYSXFI | 7702XFI codY::spc | This study |
| 7702XF5 | 7702 atxA-lacZ translational fusion | This study |
| 7CodYSXF5 | 7702XF5 codY::spc | This study |
| Plasmids | ||
| pUC1318Spc | Laboratory stock | 35 |
| pBAD | Expression vector | 21 |
| pBAK | Suicide vector in B. anthracis | 34 |
| pBALA20 | lacZ-carrying vector | 36 |
| pAT187 | Conjugative replicating plasmid used for complementation in B. anthracis; Kanr | 62 |
| pAT113 | Conjugative suicide plasmid | 61 |
| pGEMT-easy | Cloning vector | Promega |
| pQE30 | Expression vector | Qiagen |
| pTAX5 | atxA-harboring plasmid | 20 |
| pSALAC10 | lacZ-harboring plasmid | This study |
| pJP15 | pBAD derivative harboring codY under control of araBAD promoter | This study |
| pAtlac30 | pAT113 derivative harboring atxA-lacZ translational fusion | This study |
| pAtxAc4 | pQE30 derivative harboring atxA | This study |
| pCodY4 | pAT187 derivative harboring codY | This study |
| pCOD30 | pBAK derivative harboring interrupted allele of codY | This study |
| pXF1 | pAT113 derivative harboring atxA-lacZ translational fusion | J.-C. Sirard, personal communication |
| Oligonucleotides | ||
| CompCodYF | GCTGGGATCCGAAGATTTATCGTTTGAAGCATCTG | |
| CompCodYR | GGTGGGATCCGAGGAGAGTTTTATAAATTAGTTTG | |
| LacZ-N | GGATCCACCATGATTACGGATTCACTGGCCGTCGTT | |
| LacZ-C | CCCGGTTATTATTATTTTTGACACCAGACC | |
| AtxAF | GGATTCCTAACACCGATATCCATCGAAAAGGAACATATAAG | |
| AtxAR | AAGCTTGGGCATTTATATTATCTTTTTGATTTCATGAAAATCTCTTTCTGTAGG | |
| atxA-F-AC | ACTTTTGCATCCATACGTTCTTGGGCAATT | |
| atxA-R-A | GGATCCTCGGACTGTTTTATCTGCGACCTGTAGATA | |
| oPJ33 | GCGCTTTCAAAAATAGAAGAATACCTCATT | |
| oPJ36 | CCTCCTTCAGGAATATGCTACTAGTTTTAC | |
| oPJ37 | GGTATGGTAATTTTGAAAAATCTAACGCTA | |
| oPJ38 | CGACAAAAAAGTGAATTTTCACAATTCCAA | |
| qPCR-atxA1-F | ACAGGTCGCAGATAAAACAGTCC | |
| qPCR-atxA1-R | GAACAAGTAAATTCCAAGATGGAGGA | |
| qPCR-atxA2-F | TCTGCACTGCTTGAAAAACG | |
| qPCR-atxA2-R | CCATGTCTTGGAGTGATTCGTTA | |
| qPCR-tuf-F | GCCCAGGTCACGCTGACTAT | |
| qPCR-tuf-R | TCACGAGTTTGAGGCATTGG |