Table 6.
Primer sets and real-time RT-PCR conditions used for analysis of selected H. irritans ESTs.
| Unigene ID (Genbank accession number) | Gene description | Upstream/downstream primer sequences (5'-3') | PCR annealing conditions |
|---|---|---|---|
| 13_D07 (HO000820) |
5'-nucleotidase | AGTGGACAAATGTCCCGAAG AGCATTGGGGTTTGAATGAG | 55°C, 30 s |
| 6_F11 (HO000609) |
T-cell immunomodulatory protein | CCGGTGACTTTGATGGAGAT GATAATGGCTCCCCTTTGGT | 55°C, 30 s |
| 10_G05 (HO000738) |
RNase L inhibitor | GCCGATCGTGTTATTGTCCT CCGGATCGTTTTTGTTCAGT | 55°C, 30 s |
| 7_A04 (HO000619) |
Proteasome subunit beta | CAGGCGAGGTCCATTATTGT AGTGCGCGACCTCAAGTAGT | 55°C, 30 s |
| 12_H09 (HO000808) |
Proteasome maturation protein | GAGGAATCGTGAGGGTTTGA ACATGGGGTTGTCGGATAAA | 55°C, 30 s |
| 7_F08 (HO000644) |
vATPase subunit d | TGTTTTTCCGTCACCAGTCA GGCACAAACCCTCCAAGTAA | 60°C, 30 s |
| 9_A08 (HO000679) |
vATPase subunit f | TGTTGGATTCTTGCTTGGTG GGCACTGGTGATGTATGTGC | 60°C, 30 s |
| 17_H03 (HO001053) |
vATPase proteolipid subunit | GTCCAGCCAGACTGTGATGA AATCAATCGCGGACAAAAAC | 60°C, 30 s |
| 26 (HO004524) |
Ferritin light chain | TGATCATGTTGAACCCGAGA CGGCTGGTCAATTTCTTGAT | 60°C, 30 s |
| 154 (HO004652) |
Ferritin heavy chain | GTTGTTGCCCCTGCTGTATT TGAAAAGTGGGCTCCCATAG | 60°C, 30 s |
| 84 (HO004582) |
Ubiquitin-protein ligase | TCGCATCTGTTTGGATGTGT CGGGAAAACTTTTGAGTCCA | 60°C, 30 s |
| 146 (HO004644) |
Ubiquitin family (UBQ-1) | CCCGACCAACAACGTTTAAT CGACGAAGACGGTGAATTTT | 60°C, 30 s |
| 4_E04 (HO000538) |
Ubiquitin carboxyl-terminal hydrolase | AGCCAGAGATGTTGGAATGG TCGATGTAAATTGCCGCATA | 60°C, 30 s |
| 7 (HO004505) |
Vitellogenin 3 | GAGCTTTTTGCGTTGTAGCC ACAAAAGTGGGAGCAACACC | 60°C, 30 s |
| 20 (HO004518) |
Vitellogenin 1 | GAGCTTTTTGCGTTGTAGCC ACAAAAGTGGGAGCAACACC | 60°C, 30 s |
| 76 (HO004574) |
Vitellogenin 2 | ACGGCCGGTTGTGAGATTAT AGCATCTTTTTCGGTCTTGC | 60°C, 30 s |
| 2_B12 (HO000479) |
Elastase inhibitor | CAAGGGTGAATGGGAAAAGA TAAAGGCCTTCACGTTCCTG | 60°C, 30 s |
| 90 (HO004588) |
Serine protease, midgut specific trypsin, secreted | TGCGTTATATTCCGTTGGTG CTTTGTCAACGGCATAAGCA | 60°C, 30 s |
| 230 (HO004728) |
Serine protease | TGGCTACAATGAATGCAAGC GGTTAGCACCAGGGAACGTA | 60°C, 30 s |
| (FJ025436) | H. irritans 16S rRNA | TTTAAATGGCCGCAGTATCC GATTTATAGGGTCTTCTCGTCTTTT | 60°C, 30 s |