TABLE 1.
Oligonucleotides used in this study
| Namea | Use (restriction enzyme) | Sequenceb (5′-3′) |
|---|---|---|
| FLAG-0077 Fc | Amplify cbu0077 for pFLAG-0077 (BamHI) | AAGGATCCATGAGACAACTCGTTTCAATTAAA |
| FLAG-0077 Rc | Amplify cbu0077 for pFLAG-0077 (XhoI) | AAGAGCTCTTACATAATAGAACACCCACGA |
| FLAG-0077 R C-A | Amplify cbu0077 C260A (XhoI) | AAGAGCTCTTACATAATAGAAGCCCCACGA |
| FLAG-0077 R C-S | Amplify cbu0077 C260S (XhoI) | AAGAGCTCTTACATAATAGAAGACCCACGA |
| FLAG-0077 F@85 | Amplify cbu0077 from amino acid 85 (BamHI) | AAGGATCCAATGATTGGCAACTAAGAAAG |
| FLAG-0077 R@84 | Amplify cbu0077 up to amino acid 84 (XhoI) | CACTCGAGGAAAATAGTTCGAGCCGAAGT |
| FLAG-0077 R@144 | Amplify cbu0077 up to amino acid 144 (XhoI) | CCCTCGAGTTACGGATGTTTTTCAAAAAAGGC |
| pJB-0077 F | Amplify cbu0077 for pJB-Cm:3 × FLAG (SalI) | AAGGTCGACATGAGACAACTCGTTTCAATTAAA |
| pJB-0077 R | Amplify cbu0077 for pJB-Cm:3× FLAG (SalI) | TCAGTCGACTTACATAATAGAACACCCACGA |
| pJB-0077 R C-A | Amplify cbu0077 C260A (SalI) | TCAGTCGACTTACATAATAGAAGCCCCACGA |
| pJB-0077 R C-S | Amplify cbu0077 C260S (SalI) | TCAGTCGACTTACATAATAGAAGACCCACGA |
| cbu0077 up F | Amplify 2 kb upstream of cbu0077 (XbaI) | AATCTAGACGCCGCTTATTAAAAATTCC |
| cbu0077 up R | Amplify 2 kb upstream of cbu0077 | AGTATCCTTACATAAAAATCTCCTTATAATCTT |
| cbu0077 down F | Amplify 2 kb downstream of cbu0077 | GATTTTTATGTAAGGATACTGACGCAATTT |
| cbu0077 down R | Amplify 2 kb downstream of cbu0077 (SalI) | AAGTCGACGCTGTTTAATAGTTCGAACG |
| pJC-0077 F | Amplify pJC-0077 (NotI) | AGTATCCTTACATAAGCGGCCGCGCATGCAAATCTCCTTATAATCTT |
| pJC-0077 R | Amplify pJC-0077 (NotI) | GATTTTTATGTGCATGCGCGGCCGCAAGGATACTGACGCAATTT |
| KanF | Amplify kanamycin resistance cassette from pJB-Kan (NotI) | GACGCGGCCGCAGCTTATGGCTTCGTTTCGCAG |
| KanR | Amplify kanamycin resistance cassette from pJB-Kan (NotI) | GACGCGGCCGCTCAGAAGAACTCGTCAAGAAGGCG |
| Kan probe F | Amplify fragment of kanamycin resistance cassette for Southern hybridization probe | GTGGAGAGGCTATTCGGCTATG |
| Kan probe R | Amplify fragment of kanamycin resistance cassette for Southern hybridization probe | TTGAGCCTGGCGAACAGTT |
| cbu0077 probe F | Amplify 461 bp of cbu0077 for Southern hybridization probe | ATAGATTTGACGGCTGTGC |
| cbu0077 probe R | Amplify 461 bp of cbu0077 for Southern hybridization probe | AAGGGAGGACTCATCGTTGC |
| ompA qPCR F | Quantification of Coxiella genomesd | CAGAGCCGGGAGTCAAGCT |
| ompA qPCR R | Quantification of Coxiella genomesd | CTGAGTAGGAGATTTGAATCGC |