Table 1.
No | Protein/(Gene) | Primer | sequence (5′–3′) | Primer efficiency | Amplicon length (bp) | Annealing temperature (°C) |
---|---|---|---|---|---|---|
Primers designed to perform qRT-PCR for gene validation | ||||||
1 | C–C motif chemokine ligand 2 (CCL2) | CCL2- sense | CAATCAATGCCCCAGTCACCT | 94.99% | 175 | 55 |
CCL2- antisense | TCCTGAACCCACTTCTGCTTG | |||||
2 | Vascular cell adhesion molecule 1 (VCAM1) | VCAM1- sense | CCCTGAGCCCTGTGAGTTTT | 98.01% | 138 | 60 |
VCAM1- antisense | GGCCACCACTCATCTCGATT | |||||
3 | Interleukin 8, C-X-C motif chemokine ligand 8 (CXCL8) | CXCL8- sense | CTCCAAACCTTTCCACCCCA | 94.99% | 153 | 55 |
CXCL8- antisense | TTCTCCACAACCCTCTGCAC | |||||
4 | Serum amyloid A2 (SAA2) | SAA2- sense | CCTTGGTCCTGAGTGTCAGC | 96.66% | 150 | 60 |
SAA2- antisense | CATAGTTCCCCCGAGCATGG | |||||
5 | Serum amyloid A1 (SAA1) | SAA1- sense | TTTTGATGGGGCTCGGGAC | 95.02% | 150 | 60 |
SAA1- antisense | TCGGTGATCACTTCTGCAGC | |||||
6 | Interleukin 1 receptor like 1 (IL1RL1) | IL1RL1- sense | AAGGTACAGGGCGCACAAG | 93.29% | 150 | 60 |
IL1RL1- antisense | CCTTGCTCATCCTTGACCGT | |||||
7 | Cyclin-dependent kinase 6 | CDK6- sense | CCGTGGATCTCTGGAGTGTTG | 98.10% | 117 | 60 |
CDK6- antisense | GGAGTCCAATCACGTCCAAGA | |||||
8 | Collagen alpha-1 | COL12A1- sense | GACCAGCACTTCCCTCAAAGA | 95.22% | 114 | 60 |
COL12A1- antisense | GGTCGGGTAGTTTCTTGAGCA | |||||
9 | Dehydrogenase/reductase SDR family member 2 | DHRS2- sense | GCTGTCATCCTGGTCTCTTCC | 96.54% | 113 | 60 |
DHRS2- antisense | AGCTCCAATGCCAGTGTTCTA | |||||
10 | Calponin-1 | CNN1- sense | TGCGAATTCATCAATAAGCTGCA | 97.12% | 112 | 60 |
CNN1- antisense | ACTTGGTGATGGCCTTGATGA | |||||
11 | Carboxypeptidase A4 | CPA4- sense | ATTATGGAGGAAGACGCGGTC | 97.67% | 123 | 60 |
CPA4- antisense | TACACTTCGGAGCAAGGGTTG | |||||
12 | β-microglobulin (house-keeping gene) (b2m) | b2m- sense | GCTCGCGCTACTCTCTCTTT | 98.84% | 134 | 55 |
b2m- antisense | CGGATGGATGAAACCCAGACA | |||||
Primers designed to knock-out Lbp gene (SPΔLbp) | ||||||
13 | F1 fragment (351 bp upstream to Lbp gene) | Ko Lbp-F1 F | CCTTAGCCTACTTGATCCTGCT | - | 351 | 60 |
Ko Lbp-F1 R | TTCTACTCCTGAAAATTTAATCTGTCAAG | |||||
14 | Bla gene | Ko Bla F | CTTGACAGATTAAATTTTCAGGAGTAGAAATGAGTATTCAACATTTCCGTGTCG | - | 913 | 60 |
Ko Bla R | TGTCATTTTGTGGGTCTGACTCTTTACCAATGCTTAATCAGTGAGGC | |||||
15 | F2 fragment (300 bl downstream to Lbp) | Ko Lbp-F2 F | AGAGTCAGACCCACAAAATGACA | - | 300 | 60 |
Ko Lbp-F2 R | CAATTTGTTCGGCGTTGATCC |