Sequences of oligonucleotides used for chimeric plasmid construction, PCR, and gel mobility shift assays
Bold type indicates the differences of oligonucleotides of the mutated binding sites from native binding sites in the OCTN1 promoter.
Oligonucleotide | Sequence (5′ to 3′) |
|---|---|
| OCTN1 | |
| p-254 | CTCGAGAGGAATGGTAAATTCAGGATCAGG |
| p-2142 | CTCGAGAGCAATGGCAAATAGTCGAGTGG |
| p-1682 | CTCGAGTGGCTTTACCTCCCTCTCTGTTC |
| p-1263 | CTCGAGTGCAAAATCTCCAAAGAGGTGAGC |
| p-699 | CTCGAGTCATCCCTTCCAGGCTGTGG |
| p-189 | CTCGAGGGAAGCCCCCGTCAGGTCC |
| p-150 | CTCGAGGAAGCACAGGGCGGAGACAG |
| p-61 | CTCGAGTCCCAAGTGTACAGTGGCATCAAGC |
| p+92 | AAGCTTCGCTCCCCAAGGATGTTAGAACGTTCC |
| For quantity | |
| OCTN1 | ACCTCAGTGGGTTACTTTGCTC |
| GGTAGAGCTCAGCAGTGAAGAC | |
| OCTN2 | GGGCAAGTTTGGAGTCACGG |
| AGCAAGTCAGACACAGGTCAAGAG | |
| For 5′-RACE | |
| AP1 | CCATCCTAATACGACTCATATAGGGC |
| 2RDT | CGAGTCAAGTCGACGAAGTGC |
| GSP1 | CCAGGAACACGACTGACATACC |
| GSP2 | CCACTGCCGCTCCGAAAC |
| For cloning | |
| RUNX1 sense | TACCAACCAAGAAGGGGCGGA |
| RUNX1 antisense | TCTAGATTTCTGATGCATCAGAGCAGAG |
| Gel shift assay | |
| Sp1 | CCTTGGGGGCTTGCGGGCGCGCGAAGC |
| RUNX1 | CCGGGGATGGGGGTGTGGTCCCAAGTGT |
| NF-κB | TTGAAGGGAAATTCCCCCAGTCC |
| Mutagenesis | |
| Sp1 | GGTCCTTGGGGGCTTGCGGGCG |
| RUNX1 | CCGGGGATGGGGGTGTTTTCCCAAGTGT |
| NF-κB | CAACTTTGAAAATAAATTCCCCCAGTGGGC |
RACE, rapid amplification of cDNA ends.