Sequence of the oligonucleotides used in the studya
| PCR Fragments . | Sequences . | RS . |
|---|---|---|
| Recombinant proteins | ||
| T18dα1α2 | S 5′-TACCATATGGGCTCACACTCGCTGAGGTACTTCTACTTC-3′ | NdeI |
| AS 5′-CACATGGGCCTTTGGGGAATCTGTGCACTCCT-3′ | ||
| Kbbα1α2 | S 5′-GGCATATGGGCCCACACTCGCTGAGGTATTTCG-3′ | NdeI |
| AS 5′-CGTGATGGGTCACATGTGTTTTTGGAGGATCTGTGCGCAGCAGCGTCGCGTTCCC-3′ | ||
| HLA-A2 α1α2 | S 5′-GGGAATTCCATATGGGCTCTCACTCCATGAGGTATTTC-3′ | NdeI |
| AS 5′-CACATGTGTTTTTGGAGGATCCGTGCGCTGCAGCGTCTCCTT-3′ | ||
| T18dα3 (for Kb/T18d) | S 5′-GAAGAACGGGAACGCGAGGCTGCTGCGCACAGATCCTCCAAAAACACATGTG-3′ | |
| AS 5′-CCGGATCCCCATCTCAGGGTGAGAGGCTC-3′ | BamHI | |
| T18dα3 (for A2/T18d) | S 5′-CACATGTGTTTTTGGAGGATCCGTGCGCTGCAGCGTCTCCTT-3′ | |
| AS 5′-CCGGATCCCCATCTCAGGGTGAGAGGCTC-3′ | BamHI | |
| Kbdα3 | S 5′-GCACAGATTCCCCAAAGGCCCATGTG-3′ | |
| AS 5′-AGGGATCCGGATGGAGGCTCCCATC-3′ | BamHI | |
| Cell transfectants | ||
| T18dα1α2 | S 5′-CCCAAGCTTGCCACCATGAGGATGGGGACCATGGTG-3′ | HindIII |
| AS 5′-CACATGGGCCTTTGGGGAATCTGTGCACTCCT-3′ | ||
| Kbbα3 | S 5′-GCACAGATTCCCCAAAGGCCCATGTG-3′ | BamHI |
| AS 5′-GCGGGATCCTCACGCTAGAGAATGAGGCTC-3′ | ||
| Site-directed mutagenesis | ||
| T18d-G197D | 5′-GCCAGACCTGAAGATGATGTCACCCTGAGG-3′ | |
| T18d-D198K | 5′-GCCAGACCTGAAGGTAAAGTCACCCTGAGG-3′ | |
| T18d-G197D/D198K | 5′-GCCAGACCTGAAGATAAAGTCACCCTGAGG-3′ | |
| T18d-T228M | 5′-TTGATTCAGGACATGGAGCTTGTC-3′ | |
| Kd-D197G/K198D | 5′-CAGCAGACCTGAAGGCGACGTCACCCTGAGGTGC-3′ | |
| Kb-M228T | 5′-GCTGATCCAGGACACGGAGCTCGTGGAGACCAGGCC-3′ |
| PCR Fragments . | Sequences . | RS . |
|---|---|---|
| Recombinant proteins | ||
| T18dα1α2 | S 5′-TACCATATGGGCTCACACTCGCTGAGGTACTTCTACTTC-3′ | NdeI |
| AS 5′-CACATGGGCCTTTGGGGAATCTGTGCACTCCT-3′ | ||
| Kbbα1α2 | S 5′-GGCATATGGGCCCACACTCGCTGAGGTATTTCG-3′ | NdeI |
| AS 5′-CGTGATGGGTCACATGTGTTTTTGGAGGATCTGTGCGCAGCAGCGTCGCGTTCCC-3′ | ||
| HLA-A2 α1α2 | S 5′-GGGAATTCCATATGGGCTCTCACTCCATGAGGTATTTC-3′ | NdeI |
| AS 5′-CACATGTGTTTTTGGAGGATCCGTGCGCTGCAGCGTCTCCTT-3′ | ||
| T18dα3 (for Kb/T18d) | S 5′-GAAGAACGGGAACGCGAGGCTGCTGCGCACAGATCCTCCAAAAACACATGTG-3′ | |
| AS 5′-CCGGATCCCCATCTCAGGGTGAGAGGCTC-3′ | BamHI | |
| T18dα3 (for A2/T18d) | S 5′-CACATGTGTTTTTGGAGGATCCGTGCGCTGCAGCGTCTCCTT-3′ | |
| AS 5′-CCGGATCCCCATCTCAGGGTGAGAGGCTC-3′ | BamHI | |
| Kbdα3 | S 5′-GCACAGATTCCCCAAAGGCCCATGTG-3′ | |
| AS 5′-AGGGATCCGGATGGAGGCTCCCATC-3′ | BamHI | |
| Cell transfectants | ||
| T18dα1α2 | S 5′-CCCAAGCTTGCCACCATGAGGATGGGGACCATGGTG-3′ | HindIII |
| AS 5′-CACATGGGCCTTTGGGGAATCTGTGCACTCCT-3′ | ||
| Kbbα3 | S 5′-GCACAGATTCCCCAAAGGCCCATGTG-3′ | BamHI |
| AS 5′-GCGGGATCCTCACGCTAGAGAATGAGGCTC-3′ | ||
| Site-directed mutagenesis | ||
| T18d-G197D | 5′-GCCAGACCTGAAGATGATGTCACCCTGAGG-3′ | |
| T18d-D198K | 5′-GCCAGACCTGAAGGTAAAGTCACCCTGAGG-3′ | |
| T18d-G197D/D198K | 5′-GCCAGACCTGAAGATAAAGTCACCCTGAGG-3′ | |
| T18d-T228M | 5′-TTGATTCAGGACATGGAGCTTGTC-3′ | |
| Kd-D197G/K198D | 5′-CAGCAGACCTGAAGGCGACGTCACCCTGAGGTGC-3′ | |
| Kb-M228T | 5′-GCTGATCCAGGACACGGAGCTCGTGGAGACCAGGCC-3′ |
The positions of the mutated nucleotides in the primer DNA sequence are underlined. S, Sense; AS, antisense; RS, restriction site.