Target gene
|
Primer
|
Sequence a(5′-3′)
|
Primer locationb
|
Restriction site
|
Expected product (bp)
|
---|
Porcine IL-6
|
IL6fs
|
GACGAATTCATGAACTCCCTCTCCACA
|
1–18
|
EcoR I
|
639
|
IL6rs
|
GTCGTTAAC CTACATTATCCGAATG
|
471–488
|
Hpa I
| |
PPV VP2
|
VP2fs
|
ACCATGGGGGGGGTTGGTG
|
2739–2758
| |
1635
|
VP2rs
|
ATCCTATTTCTAGTATAATTT
|
4359–4373
| | |
PRV gH
|
gH fs
|
GCGTGTACTGCGACTGCGTGTT
|
62,700–62,721
| |
355
|
gH rs
|
CGACCTGGCGTTTATTAACCGAGA
|
63,032–63,055
| | |
PPV NS1
|
NS1fs
|
CCAAAAATGCAAACCCCAATA
|
1808–1828
| |
194
|
NS1rs
|
TCTGGCGGTGTTGGAGTTAAG
|
1981–2001
| | |
- aThe restriction enzyme sites used for the construction are underlined
- bThe primers IL6fs/IL6rs were designed to amplify porcine IL-6 gene using plasmid pIRES-IL6 as templates. The primers VP2fs/VP2rs and NS1fs/NS1rs were designed based on the nucleotide sequence (KF429255) of PPV HN genome to amplify VP2 and NS1genes, respectively. The primers gH fs/gH rs were designed to amplify gH gene based on PRV gH nucleotide sequence (KP257591) retrieved from the GenBank