Primers used to amplify DNA product for the ornithine–urea cycle(O–UC) gene in gulf toadfish (Opsanus beta) and gene specific primers designed for quantitative (qPCR) used in this study
| Primer . | Accession no. . | Sequence (5′→3′) . | Product size (bp) . |
|---|---|---|---|
| OCT–Fc | XM_001334635c | GTCYATTGCCACGATHTTTGA | 582 |
| OCT–Rc | TTTCTCGTCTTCTTGBCCCAT | ||
| ASS–Fc | NM_001004603c | TATGCTGGTCTGGCTGAAGGA | 939 |
| ASS–Rc | ACACTCGGGGTTGARCCAGAA | ||
| ASL–Fc | NM_20045c | CAATGAACGCAGGCTSAAGGA | 702 |
| ASL–Rc | TAGGTCTTCGTTGTANGTGCT | ||
| ARG*e5f1a | NM_001045197a | TTGGGCTTAGAGAYGTGGAYC | 240 |
| ARG*e7r2a | CCTTCTCTGTARGTNAGTCCTCC | ||
| EF1α–qFb | NM_131263b | AGGTCATCATCCTGAACCAC | 140 |
| EF1α–qRb | GTTGTCCTCAAGCTTCTTGC | ||
| GS–qFd | AF118103d | ACTCGCTGCCATACAAACTT | 184 |
| GS–qRd | TCGTGGATGTTTGAGGTTTC | ||
| CPS–qFe | AF169248e | ATGGCAATCAAAGTCGTTCG | 252 |
| CPS–qRe | CCAGACTAAACTCAAAGCAG | ||
| OCT–qFf | EU704512 | TGAAAAGAGAAGCACCAGAAC | 171 |
| OCT–qRf | ATACACTCGTGCCAAGACAA | ||
| ASS–qFf | EU704511f | CCAATGCCGTTTATGAGGAC | 150 |
| ASS–qRf | TCAAAGCGTCCCTGGTCATT | ||
| ASL–qFf | EU704510f | AGTCGGAACAGTATGGATGC | 173 |
| ASL–qRf | AAACTGCTGCCTGTGCTGTA | ||
| ARG–qFf | EU704513f | TGATTGAAGGCGTACTCCAT | 145 |
| ARG–qRf | TGGATAACGGTGGGGTCAA |
| Primer . | Accession no. . | Sequence (5′→3′) . | Product size (bp) . |
|---|---|---|---|
| OCT–Fc | XM_001334635c | GTCYATTGCCACGATHTTTGA | 582 |
| OCT–Rc | TTTCTCGTCTTCTTGBCCCAT | ||
| ASS–Fc | NM_001004603c | TATGCTGGTCTGGCTGAAGGA | 939 |
| ASS–Rc | ACACTCGGGGTTGARCCAGAA | ||
| ASL–Fc | NM_20045c | CAATGAACGCAGGCTSAAGGA | 702 |
| ASL–Rc | TAGGTCTTCGTTGTANGTGCT | ||
| ARG*e5f1a | NM_001045197a | TTGGGCTTAGAGAYGTGGAYC | 240 |
| ARG*e7r2a | CCTTCTCTGTARGTNAGTCCTCC | ||
| EF1α–qFb | NM_131263b | AGGTCATCATCCTGAACCAC | 140 |
| EF1α–qRb | GTTGTCCTCAAGCTTCTTGC | ||
| GS–qFd | AF118103d | ACTCGCTGCCATACAAACTT | 184 |
| GS–qRd | TCGTGGATGTTTGAGGTTTC | ||
| CPS–qFe | AF169248e | ATGGCAATCAAAGTCGTTCG | 252 |
| CPS–qRe | CCAGACTAAACTCAAAGCAG | ||
| OCT–qFf | EU704512 | TGAAAAGAGAAGCACCAGAAC | 171 |
| OCT–qRf | ATACACTCGTGCCAAGACAA | ||
| ASS–qFf | EU704511f | CCAATGCCGTTTATGAGGAC | 150 |
| ASS–qRf | TCAAAGCGTCCCTGGTCATT | ||
| ASL–qFf | EU704510f | AGTCGGAACAGTATGGATGC | 173 |
| ASL–qRf | AAACTGCTGCCTGTGCTGTA | ||
| ARG–qFf | EU704513f | TGATTGAAGGCGTACTCCAT | 145 |
| ARG–qRf | TGGATAACGGTGGGGTCAA |
Accession numbers for Danio rerio and O. beta sequences from which the primers were designed are provided. Abbreviations: Elongation factor 1-alpha (EF1α); glutamine synthetase (GS); carbamoyl phosphate synthetase (CPS); argininosuccinate synthetase (ASS); argininosuccinate lyase(ASL); arginase (ARG); F1 forward primer (F); reverse primer (R); quantitative PCR primer (q)
Primers used from Wright et al. (Wright et al., 2004) designed for Oncorhynchus mykiss
Primers designed by Edward M. Mager (University of Miami) designed for Pimephales promelas
Degenerate primers designed for O. beta from conserved nucleotide sequences across multiple vertebrate species, including zebrafish (D. rerio), human (Homo sapiens), mouse (Mus musculus) and rat (Rattus norvegicus)
Primers designed from published sequence of O. beta in Walsh et al. (Walsh et al., 1999)
Primers designed from published sequence of O. beta in Kong et al.(Kong et al., 2000)
Primers designed from sequence of O. beta obtained in this study