Table 1.
Designed primers used for PCR and DNA sequencing (Eurogentec S.A., Belgium)
| Primer | Sequence (5′→3′) | Position (relative to human BSEP [NM_003742]) |
|---|---|---|
| Oligo(dT)-anchor |
5′-GACCACGCGTATCGATGTCGACTTTTTTTTTTTTTTTT |
|
| PCR-anchor |
5′-GACCAGGCGTATCGATGTCGAC |
|
| BSEP_RACE_A |
5′-AGTGTTGTTTGCCTGTAGC |
3615-3633 |
| BSEP_RACE_B |
5′-TGAAGAAAATCGAAACAAT |
273-291 |
| BSEP_RACE_C |
5′-TGTGCCAAAAATGAGGAG |
367-381 |
| BSEP_RACE_D |
5′-AGTTCTTGTAATTCAGTGTCA |
405-425 |
| BSEP_Af |
5′-CTCGACCTGATACGCAAGTTCTGA |
3392-3415 |
| BSEP_Ar |
5′-AATGGCCCGAGCAATAGCAATAC |
3805-3825 |
| BSEP_Bf |
5′-CAAGGGAAGGTGATGATAGATGG |
3526-3548 |
| BSEP_Br |
5′-GATGGGGGCTCCTGTGGTAACT |
4065-4086 |
| BSEP_Cf |
5′-TGTGCTTCTTCCCCTTCTTGGCT |
2840-2862 |
| BSEP_Cr |
5′-TGCCCATCTATCATCACCTTCC |
3530-3548 |
| BSEP_Df |
5′-CAACGCTCCAAGTCTCA |
2215-2231 |
| BSEP_Dr |
5′-GTGCGGATATTACTGAG |
2956-2972 |
| BSEP_Ef |
5′-CAAGGCTTGCTACGGATGC |
2702-2720 |
| BSEP_Er |
5′-TGATTGGGGGTTGTCGATC |
3295-3310 |
| BSEP_Ff |
5′-GTGGTGGCCAGAAACAAAG |
1805-1823 |
| BSEP_Fr |
5′-TCTGGCTGAATAAAAAGGCA |
2442-2461 |
| BSEP_Gf |
5′-TGGATCGAATTAAGGGTGAA |
1364-1383 |
| BSEP_Gr |
5′-GCGATGAGCAACCGAAATGA |
1955-1974 |
| BSEP_Hf |
5′-GATGGGATTCTTTACTGGATTC |
1089-1110 |
| BSEP_Hr |
5′-CCTTCACTGGGGTCATAGAA |
1537-1556 |
| BSEP_If |
5′-GCAGCTCGTCAGATACAGAA |
625-644 |
| BSEP_Ir |
5′-CAGAAGGCCAATGCATAACA |
1132-1151 |
| BSEP_Jf |
5′-TGAAGGCCTATGCCAAAGC |
938-956 |
| BSEP_Jr |
5′-TTCACCCTTAATTCGATCCA |
1364-1383 |
| BSEP_Kf |
5′-ATTGTTTCGATTTTCTTCA |
273-291 |
| BSEP_Kr |
5′-GCTTTGGCATAGGCCTTCA |
938-956 |
| BSEP_Lf |
5′-AACCCTTGTCCAGATTTTCCTC |
1197-1218 |
| BSEP_Lr |
5′-ATGATGGGTTTCCGGTCTATTG |
1313-1334 |
| FullBsep_felinef |
5′-TTGCAATTACCATGTCTGACTCAGTAATTCTTCGC |
|
| FullBsep_feliner |
5′-TCAACTGATGGGGGCTCCTGTGATGACTAG |
|
| FullBsep_caninef |
5′-TTGCAATTACCATGTCTGATGCAGTAATTCTTCGC |
|
| FullBsep_caniner |
5′-TCAACTGATGGGGGCTCCTGTGGTAACTAG |
|
| FullBSEP + AttB1_humanf |
5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCGCCA CCATGTCTGACTCAGTAATTCTTC |
|
| FullBSEP + AttB1_humanr |
5′-GGGGACCACTTTGTACAAGAAAGCTGGGTCTCAAC TGATGGGGGATCCAGTG |
|
| AttB1_felinef |
5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCGCC ACCTTGCAATTACCATGTCTGACTCAGTAATTCTTCGC |
|
| AttB1_feliner |
5′-GGGGACCACTTTGTACAAGAAAGCTGGGTCTCAAC TGATGGGGGCTCCTGTGATGACTAG |
|
| AttB1_caninef |
5′-GGGGACAAGTTTGTACAAAAAAGCAGGCTTCGCC ACCTTGCAATTACCATGTCTGATGCAGTAATTCTTCGC |
|
| AttB1_caniner | 5′-GGGGACCACTTTGTACAAGAAAGCTGGGTCTCAAC TGATGGGGGCTCCTGTGGTAACTAG |
The Oligo(dT)-anchor, PCR-anchor and the specific primer BSEP_RACE_A, were used for 3′RACE-PCR. The BSEP_Ar, Oligo(dT)-anchor, PCR-anchor, BSEP_RACE_B, C and D were used for 5′RACE-PCR. Each primer pair is denoted with “f” for the forward primer, and “r” for the reverse primer.