Table 1 Primers and probes.
Targeted gene | Primer/probe name | Sequence (5’- 3’) | Accession n°/ reference | ||
|---|---|---|---|---|---|
Primers for PCR and RT-PCR | PCR | RT-PCR | |||
CyHV-3 ORF3 | ORF3InF | • | TATGCCCACATGATGCTGTT | DQ657948 | |
ORF3InR | • | CAGTCAGACCCTTCCTCTGC | |||
CyHV-3 ORF55 | ORF55InF | • | AGCGCTACACCGAAGAGTCC | ||
ORF55stopR | • | TCACAGGATAGATATGTTACAAG | |||
ORF55ATGF | • | ATGGCTATGCTGGAACTGG | |||
ORF55InR | • | GGCGCACCCAGTAGATTATG | |||
CyHV-3 ORF78 | ORF78InF | • | TGGACGACGAACACCCTTC | ||
ORF78InR | • | GGTAGAGGGTACAACCACG | |||
CyHV-3 ORF132 | ORF132InF | • | GGATCCGTTTTCTGGGTCTG | ||
ORF132InR | • | CTCAATCCCTCACCGACCTC | |||
CyHV-3 ORF133 | ORF133InF | • | GACGAGATCCCTATCCGCAG | ||
ORF133InR | • | GACCTCGGGTATGGTCGGTA | |||
CyHV-3 ORF134 | ORF134stopF | • | TCAATGTTTGCGCTTGGTTTTC | ||
ORF134ATGR | • | ATGTTCCTTGCAGTGCTAC | |||
ORF134InF | • | GGTTTCTCTTTGTAGTTTTCCG | |||
ORF134InR | • | CACCCCAACTTTTGAGACAAC | |||
ORF134outseqF | • | GTCAACATGGACGAGCGTGA | |||
ORF134outseqR | • | GTGGGGATATCAAACACGCA | |||
CyHV-3 ORF135 | ORF135InF | • | ACACCACCAACGAGACATGC | ||
ORF135InR | • | CTTTTCGGACCAGAAGACCG | |||
Carp β-actin | Actin-F | • | ATGTACGTTGCCATCCAGGC | M24113 | |
Actin-R | • | GCACCTGAACCTCTCATTGC | |||
Primers for amplification of recombination cassettes | |||||
H1-gal K-H2 cassette | 134 gal K F | ATGTTCCTTGCAGTGCTACTAACCG | Warming et al. [34] | ||
CGACCATCTTCTTCGAGGCTCGGGG | |||||
CCTGTTGACAATTAATCATCGGCA | |||||
134 gal K R | TCAATGTTTGCGCTTGGTTTTCATG | ||||
TTCTTGACGTCTTTTGCGACCAGGA | |||||
TCAGCACTGTCCTGCTCCTT | |||||
H1-ORF134-H2 | H1F | GCTCATCAATCGCAGCAGCA | DQ657948 | ||
cassette | |||||
H2R | CAAGCCATTATCCTGTTGGG | ||||
Primers and probes for real-time TaqMan PCR quantification of CyHV-3 genome | |||||
CyHV-3 ORF89 | KHV-86F | GACGCCGGAGACCTTGTG | AF411803 | ||
KHV-163R | CGGGTTCTTATTTTTGTCCTTGTT | ||||
KHV-109P | (6FAM)CTTCCTCTGCTCGGCGAGCACG(BHQ1) | ||||
Carp glucokinase | CgGluc-162F | ACTGCGAGTGGAGACACATGAT | AF053332 | ||
CgGluc-230R | TCAGGTGTGGAGCGGACAT | ||||
CgGluc-185P | (6FAM)AAGCCAGTGTCAAAATGCTGCCCACT(BHQ1) | ||||
Primers for RT-qPCR analysis of carp gene expression | |||||
40S | 40S-F | CCGTGGGTGACATCGTTACA | AB012087 | ||
40S-R | TCAGGACATTGAACCTCACTGTCT | ||||
IL-1β | IL-1β-F | AAGGAGGCCAGTGGCTCTGT | AJ245635 | ||
IL-1β-R | CCTGAAGAAGAGGAGGCTGTCA | ||||
TNF-α1 and 2 | TNF-α1 and 2-F | GCTGTCTGCTTCACGCTCAA | AJ311800 | ||
TNF-α1 and 2-R | CCTTGGAAGTGACATTTGCTTTT | ||||
CXCa | CXCa-F | CTGGGATTCCTGACCATTGGT | AJ421443 | ||
CXCa-R | GTTGGCTCTCTGTTTCAATGCA | ||||
IL-10 | IL-10-F | CGCCAGCATAAAGAACTCGT | AB110780 | ||
IL-10-R | TGCCAAATACTGCTCGATGT | ||||
IFNγ-2 | IFNγ-2-F | TCTTGAGGAACCTGAGCAGAA | AM168523 | ||
IFNγ-2-R | TGTGCAAGTCTTTCCTTTGTAG | ||||
IL-6 | IL-6M17-F | CACATTGCTGTGAGGGTGAA | AY102633 | ||
IL-6M17-R | GCATCCATAGGCTTTCTGCT | ||||