Table 3.
Oligonucleotide primers and PCR conditions for the characterization of subolesin and pathogen-specific gene expression.
| Gene descriptiona | Upstream/downstream primer sequences (5'-3') | PCR annealing conditions |
|---|---|---|
| D. variabilis subolesin [9] | CCAGCCTCTGTTCACCTTTC CCGCTTCTGAATTTGGTCAT |
54°C, 30 sec |
| R. microplus subolesin [9] | CACAGTCCGAGTGGCAGAT GATGCACTGGTGACGAGAGA |
55°C, 30 sec |
| A. marginale msp4 [29] | GGGAGCTCCTATGAATTACAGAGAATTGTTTAC CCGGATCCTTAGCTGAACAGGAATCTTGC |
60°C, 1 min |
| A. phagocytophilum msp4 [9] | GACGTGCTGCACACAGATTT CTCATCAAATAGCCCGTGGT |
54°C, 1 min |
| E. canis 16S (M73221) | GTGGCAGACGGGTGAGTAAT GCTGATCGTCCTCTCAGACC |
57°C, 30 sec |
| B. subtilis dal [35] | AATTGAAAGGGACCGACATC- TTAATGGTTTCGAGCCTTCC |
59°C, 30 sec |
| E. coli dxs [36] | CGAGAAACTGGCGATCCTTA CTTCATCAAGCGGTTTCACA |
60°C, 30 sec |
| P. pastoris CTA 1 (AB472085) | CCTGAAGGACGCCAATATGT GCTTTCCAGCCTCTTCATTG |
57°C, 30 sec |
| Tick 16S rRNA [9] | GACAAGAAGACCCTA ATCCAACATCGAGGT |
42°C, 30 sec |
aWhen published, references are shown for oligonucleotide sequences. When designed for this study, GenBank accession numbers are shown in parenthesis.