TABLE 1.
Primer sequences used in this studya
| Primer | Sequence (length [bp]) |
|---|---|
| Genomic | |
| C.B.1b | 5′-ACT CAA CGC ACT GGA ACC GC-3′ |
| C.B.2b | 5′-TAG CTG AAG CCA ATT CGC C-3′ (257 bp) |
| G4131b | 5′-CTG ATG TGT CAA GTA ATG TCG G-3′ |
| G4132b | 5′-GTT CAT GGT TAT GAT TCT GCG-3′ (183 bp) |
| 16S1c | 5′-CTC CTG GCG GCG AGA GTG GC-3′ |
| 16S2Nc | 5′-GTT AGC TTC GCT ACT AAG AAG GGA ACT TCC C-3′ (779 bp) |
| Plasmidic | |
| QpRS01d | 5′-CTCGTACCCAAAGACTATGAATATATCC-3′ |
| QpRS02d | 5′-CACATTGGGTATCGTACTGTCCCT-3′ (363 bp) |
| QpH11d | 5′-TGACAAATAGAATTTCTTCATTTTGATG-3′ |
| QpH12d | 5′-GCTTATTTTCTTCCTCGAATCTATGAAT-3′ (1,042 bp) |
| Hfrag1e | 5′-ATT GCT ATC ACT GAG GGT GAC G-3′ |
| Hfrag2e | 5′-CTG ACG AAG AAG CAG CAT TAG C-3′ (508 bp) |
| HF1e | 5′-TCC TAA ACA AGT GAT GGT CTC C-3′ |
| HF2e | 5′-TTC GCA GAA GTC AGC TAT GC-3′ (183 bp) |
a
Cycling conditions were as follows: 94°C for 5 min with 35 (total) cycles of denaturation (at 94°C for 30 min), annealing (at 55°C for 30 min), and extension (at 72°C for 30 min). After the 35 cycles, the PCR product was held for 10 min at 72°C.
b
Primers CB.1 and CB.2 were derived from the C. burnetii superoxide dismutase gene, and primers G4131 and G4132 were derived from a shotgun HindIII subcloning fragment (297 bp).
c
Primers 16S1 and 16S2N are based on DNA sequences of 16S rRNA.
d
Primers QpRS01 and QpRS02 are QpRS specific, and primers QpH11 and QpH12 are QpH1 and QpDG specific.
e
Primers Hfrag1, Hfrag2, HF1, and HF2 were used for nested PCR.