Table 1.
Gene-specific primers used for quantitative real-time PCR in this study.
| Gene | Primer | GenBank accession no. | Tm (°C) | Amplicon length (bp) | Primer Sequence (5′-3′) | |
|---|---|---|---|---|---|---|
| Forward primer | Reverse primer | |||||
| IMMUNOGLOBULINS AND THEIR RECEPTOR | ||||||
| IgT | IgT-F/IgT-R | AY870264 | 58 | CAGACAACAGCACCTCACCTA | GAGTCAATAAGAAGACACAACGA | |
| IgM | IgM-F/IgM-R | S63348.1 | 58 | AAGAAAGCCTACAAGAGGGAGA | CGTCAACAAGCCAAGCCACTA | |
| IgD | IgD-F/IgD-R | AY748802.1 | 58 | 138 | CAGGAGGAAAGTTCGGCATCA | CCTCAAGGAGCTCTGGTTTGGA |
| pIgR | pIgR-F/pIgR-R | FJ940682.1 | 58 | 145 | GTACAGCAGGTGTTCACAGTAAC | CCACAGACAGACCTTGGATAAC |
| COMPLEMENT AND PATHWAY RELATED GENES | ||||||
| Complement 3 | C3-F/C3-R | XM_021561577.1 | 58 | 213 | CCTCACAACAAGAGTGCACATC | CCAAGTGGGCAAACTCATCTCC |
| Complement 1q-2 | C1-F/C1-R | XM_021624859.1 | 58 | 176 | TGAACAACCTGAACACCCC | CAGCCTATTAGCCTGTAACTCC |
| Complement 7-1 | C7-F/C7-R | NM_001124618.1 | 58 | 218 | TATCTTCACTGCCACGGTC | TAGCCTGTAACTCCACATAGAC |
| CYTOKINES | ||||||
| interleukin 2 | IL2-F/IL2-R | AM422779.1 | 58 | 125 | TGTCTACAAGGAAACCCAA | GCTGCAACAATGCAACTAT |
| interleukin 11 | IL11-F/IL11-R | NM_001124382.1 | 58 | 191 | CAGAGCGTCAAGGAAACAC | GCTCCTGGGAAGACTGTAA |
| interleukin 22 | IL22-F/IL22-R | AM 748537 | 58 | 101 | GCAGGACAAGCATCATCCTAA | CATTGTAGTGTTTAATAGCACAGC |
| interferon α | IFNα-F/IFNα-R | XM_021578986.1 | 58 | 180 | CAGAGCCTCAGGAAGAACT | CAAGGGGTAGAAGAGCATA |
| IRF 1-2 | IRF1-F/IRF1-R | NP_001239293.1 | 58 | 173 | CGAGACTACACCAGACCCTA | TTGCTTTTGACCTCTTGTTATT |
| TNF α | TNFα-F/TNFα-R | XM_021576327.1 | 58 | 204 | GTATGCGATGACACCTGAA | GCCCCATTAGAGTGCCTTA |
| CCL 19 | CCL19-F/CCL19-R | AIN40032.1 | 58 | 139 | GTTTCCCTCGCCACTTCAA | GCCACCCACTTGCTCTTTG |
| ICHTHYOPHTHIRIUS MULTIFILIIS | ||||||
| 18S rRNA | Ic18-F/Ic18-R | 60 | AGTGACAAGAAATAGCAAGCCAGGAG | ACCCAGCTAAATAGGCAGAAGTTCAA | ||
| FLAVOBACTERIUM COLUMNARE | ||||||
| 16S rRNA | Fc16-F/Fc16-R | EU395796.1 | 60 | 146 | GAGTGGCTAAGCGAAAGTGAT | ACCTGACACCTCACGGCAC |
| REFERENCE GENE | ||||||
| EF-1α | EF1-F/EF1-R | XM_021571866.1 | 58 | CAACGATATCCGTCGTGGCA | ACAGCGAAACGACCAAGAGG | |
| β-actin | BA-F/BA-R | AB196465 | 60 | 225 | CCGCGACCTCACAGACTACC | GTGCCCATCTCCTGCTCAAA |
| OTHERS | ||||||
| cathelicidin 1 | CATH1-F/CATH1-R | NM_001124480.1 | 58 | 152 | CTGGAGGCAAGCAACAAC | CCCCCAAGACGAGAGACA |
| COL1α | COL1α-F/COL1α-R | NM_001124206.1 | 58 | 190 | ATGTGCGACGAAGTGATTTG | CGTTACCTGGAACACCCTC |
| m-csfr | CSFR-F/CSFR-R | NM_001124739.1 | 58 | 120 | CCCGCCTGTCACCCAATCT | CGTCCCACCAATGCTTCT |
| C-type lectin 4E | CTL4-F/CTL4-R | XM_021562202.1 | 58 | 137 | GCAGCCACCTTACCATC | CACCCATCTCCAATCCC |
| DnajC3 | DC3-F/DC3-R | XM_021597283.1 | 58 | 166 | TATTGAAACGTGTGTGTGGG | AGGTGGTAGTAGATGGTGCTG |
| DnajB9 | DB9-F/DB9-R | XM_021577464.1 | 58 | 234 | CCGCTGTTGTTAGTGCTTT | CGTCAGGACTCTTGTTTCG |
| ENO | ENO-F/ENO-R | XM_021578347.1 | 58 | 214 | TCAAGTCACCTGACGACCCT | ACAGCCTGCTGGATACGC |
| FHL1 | FHL1-F/FHL1-R | XM_021618853.1 | 58 | 212 | CGGCTCTGAGAATGTGG | ATGGCTGCTCCTGGTAG |
| GAPDH | GAPDH-F/GAPDH-R | NM_001124209.1 | 58 | 100 | GACACCCACTCCTCCATCTT | TGCTGTAGCCAAACTCATT |
| LAMβ3 | LAMβ3-F/LAMβ3-R | XM_021570344.1 | 58 | 211 | TCAGGGTTCGGAGGTCGTA | CACAGTCGGGAAAGGAGTAA |
| LDH | LDH-F/LDH-R | XM_021605689.1 | 58 | 105 | ACTCCAGCGTGCCCGTAT | CGTGCTTCCAGTCCTCCTT |
| PFKA | PFKA-F/PFKA-R | XM_021609920.1 | 58 | 103 | CGACACCCGTTCCACAAT | AGCCATCACAGCCTCCAC |
The primers of IgT, IgM, and EF1α were obtained from the previous study of Tacchi et al. (24).