TABLE 3.
Oligonucleotide primers and restriction genome libraries used for genome walking of the ehrlichial citrate synthase gene
| Target species | Primer | Sequence (5′-3′)b | Restriction genomic library(ies) |
|---|---|---|---|
| HGE agent | HG-SPF | CGG-ATC-AAT-GCG-ACT-TGC-G | PvuII |
| HG-SPR | TTG-CTG-AGA-TTG-GTT-CAC-C | PvuII | |
| A. marginale South Idaho | MAR-SPR | GCC-ATT-GGG-TGG-GCA-TCA | DraI |
| MAR-SPF1 | ATG-CTT-GAG-GAA-ATT-GGT-CGC-C | DraI | |
| MAR-SPF2a | CCA-GTG-AAC-ATG-TTT-ACC-GCG | DraI | |
| A. centrale | CENT-SPR1 | GAT-GAG-CAA-CGC-TGC-AGA-ACC | ScaI, DraI |
| CENT-SPR2a | CCA-CCT-GTC-GTG-ACA-AGA-TC | ScaI, DraI | |
| CENT-SPF1 | TTG-AGT-GCA-GGT-GCT-GCA-AC | PvuII | |
| CENT-SPF2a | TTT-GAG-CTT-GAA-AGA-GTG-GCC | PvuII | |
| E. chaffeensis | F3R | CAT-AGG-ATG-TGA-ATC-T | DraII |
| CHAF-SPRa | ATT-TTG-CAA-ATC-CTC-AAG-ACC | DraII | |
| F1 | CAT-GAR-CAR-AAT-GCT-TC | EcoRV | |
| CHAF-SPFa | TGT-GAT-TAA-TAT-GTT-AAT-GGC | EcoRV | |
| E. canis | CAN-SPR1 | TAA-CTT-TAT-TTC-CAT-TAG-TAT-CAC | EcoRV, DraI |
| CAN-SPR2a | CGG-TAA-TTT-CAC-TTT-TTG-ACC | EcoRV, DraI | |
| CAN-SPF | GTG-GTA-TGA-GAT-GGT-GGC-AGA-TAA-G | DraI | |
| E. muris | MUR-SPR1 | TAA-ATC-TAC-TAT-GTT-ATG-TCC | EcoRV |
| MUR-SPR2a | TCA-TAA-GTT-AAA-ACT-CCT-GTG-TC | EcoRV | |
| MUR-SPF1 | AAT-GAC-AAT-TGA-AAA-ACC-AAG | DraI | |
| ER-R1Fa | ATG-GGT-TTT-GGT-CAT-AGA-G | DraI | |
| C. ruminantium | R1b | CGA-TGA-CCA-AAA-CCC-AT | DraI |
| CR-SPRa | TAA-ATT-CCT-GAA-GTT-GCT-CAG-CAT | DraI | |
| F1b | GAT-CAT-GAR-CAR-AAT-GCT-TC | PvuII | |
| CR-SPFa | GGA-TTC-CAG-TTA-AAA-TGT-TTA-CG | PvuII | |
| E. sennetsu | R1b | CGA-TGA-CCA-AAA-CCC-AT | DraII, StuI |
| SEN-SPR1a | CTG-TAG-CCG-CAG-AAC-ATG-CC | DraII, StuI | |
| SEN-SPR2 | TTG-CAT-GGA-GCA-GTT-TTT-GC | EcoRV, ScaI | |
| SEN-SPR3a | GCA-TAC-CCT-GGA-TCA-TAA-AAG | EcoRV, ScaI | |
| SEN-SPF1 | GAG-AGG-TAT-AGT-TGA-AAG-CG | EcoRV | |
| SEN-SPF2a | AAG-GGA-AGA-GTA-CTT-TCT-GAG-TC | EcoRV | |
| SEN-SPF4 | GAG-TGA-CAG-TGA-ACA-AAA-AC | ScaI, StuI | |
| SEN-SPF5a | GCA-GAC-CTA-GAC-AAA-TTT-ACG | ScaI, StuI | |
| E. risticii | RIS-SPR | CGA-GCG-AAG-CAA-GTG-CAC-A | EcoRV, ScaI, DraI, PvuII |
| RIS-SPF | GAG-AGG-TAT-CAT-TGA-AAG-CGG | EcoRV, ScaI | |
| N. helmintoeca | NEO-SPR1 | CCA-TGG-GAT-GTG-CAT-C | EcoRV |
| NEO-SPR2a | TTT-TGC-GCG-GAC-AGG-GTC | EcoRV | |
| F1 | CAT-GAR-CAR-AAT-GCT-TC | EcoRV | |
| NEO-SPFa | GAG-AAA-GTC-CTG-CAC-ATG-CTG | EcoRV |
a
Used in the nested PCR.
b
R, A or G.