Table 3.

Oligonucleotide primers and restriction genome libraries used for genome walking of the ehrlichial citrate synthase gene

Target speciesPrimerSequence (5′-3′)bRestriction genomic library(ies)
HGE agentHG-SPFCGG-ATC-AAT-GCG-ACT-TGC-G PvuII
HG-SPRTTG-CTG-AGA-TTG-GTT-CAC-C PvuII
A. marginale South IdahoMAR-SPRGCC-ATT-GGG-TGG-GCA-TCA DraI
MAR-SPF1ATG-CTT-GAG-GAA-ATT-GGT-CGC-C DraI
MAR-SPF2a CCA-GTG-AAC-ATG-TTT-ACC-GCG DraI
A. centrale CENT-SPR1GAT-GAG-CAA-CGC-TGC-AGA-ACC ScaI,DraI
CENT-SPR2a CCA-CCT-GTC-GTG-ACA-AGA-TC ScaI,DraI
CENT-SPF1TTG-AGT-GCA-GGT-GCT-GCA-AC PvuII
CENT-SPF2a TTT-GAG-CTT-GAA-AGA-GTG-GCC PvuII
E. chaffeensis F3RCAT-AGG-ATG-TGA-ATC-T DraII
CHAF-SPRa ATT-TTG-CAA-ATC-CTC-AAG-ACC DraII
F1CAT-GAR-CAR-AAT-GCT-TC EcoRV
CHAF-SPFa TGT-GAT-TAA-TAT-GTT-AAT-GGC EcoRV
E. canis CAN-SPR1TAA-CTT-TAT-TTC-CAT-TAG-TAT-CAC EcoRV,DraI
CAN-SPR2a CGG-TAA-TTT-CAC-TTT-TTG-ACC EcoRV,DraI
CAN-SPFGTG-GTA-TGA-GAT-GGT-GGC-AGA-TAA-G DraI
E. muris MUR-SPR1TAA-ATC-TAC-TAT-GTT-ATG-TCC EcoRV
MUR-SPR2a TCA-TAA-GTT-AAA-ACT-CCT-GTG-TC EcoRV
MUR-SPF1AAT-GAC-AAT-TGA-AAA-ACC-AAG DraI
ER-R1Fa ATG-GGT-TTT-GGT-CAT-AGA-G DraI
C. ruminantium R1bCGA-TGA-CCA-AAA-CCC-AT DraI
CR-SPRa TAA-ATT-CCT-GAA-GTT-GCT-CAG-CAT DraI
F1bGAT-CAT-GAR-CAR-AAT-GCT-TC PvuII
CR-SPFa GGA-TTC-CAG-TTA-AAA-TGT-TTA-CG PvuII
E. sennetsu R1bCGA-TGA-CCA-AAA-CCC-AT DraII,StuI
SEN-SPR1a CTG-TAG-CCG-CAG-AAC-ATG-CC DraII,StuI
SEN-SPR2TTG-CAT-GGA-GCA-GTT-TTT-GC EcoRV,ScaI
SEN-SPR3a GCA-TAC-CCT-GGA-TCA-TAA-AAG EcoRV,ScaI
SEN-SPF1GAG-AGG-TAT-AGT-TGA-AAG-CG EcoRV
SEN-SPF2a AAG-GGA-AGA-GTA-CTT-TCT-GAG-TC EcoRV
SEN-SPF4GAG-TGA-CAG-TGA-ACA-AAA-AC ScaI,StuI
SEN-SPF5a GCA-GAC-CTA-GAC-AAA-TTT-ACG ScaI,StuI
E. risticii RIS-SPRCGA-GCG-AAG-CAA-GTG-CAC-A EcoRV,ScaI, DraI, PvuII
RIS-SPFGAG-AGG-TAT-CAT-TGA-AAG-CGG EcoRV,ScaI
N. helmintoeca NEO-SPR1CCA-TGG-GAT-GTG-CAT-C EcoRV
NEO-SPR2a TTT-TGC-GCG-GAC-AGG-GTC EcoRV
F1CAT-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.