Table I.

Sequence of the oligonucleotides used in the studya

PCR FragmentsSequencesRS
Recombinant proteins
T18dα1α2S 5′-TACCATATGGGCTCACACTCGCTGAGGTACTTCTACTTC-3′NdeI
AS 5′-CACATGGGCCTTTGGGGAATCTGTGCACTCCT-3′
Kbbα1α2S 5′-GGCATATGGGCCCACACTCGCTGAGGTATTTCG-3′NdeI
AS 5′-CGTGATGGGTCACATGTGTTTTTGGAGGATCTGTGCGCAGCAGCGTCGCGTTCCC-3′
HLA-A2 α1α2S 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
Kbdα3S 5′-GCACAGATTCCCCAAAGGCCCATGTG-3′
AS 5′-AGGGATCCGGATGGAGGCTCCCATC-3′BamHI
Cell transfectants
T18dα1α2S 5′-CCCAAGCTTGCCACCATGAGGATGGGGACCATGGTG-3′HindIII
AS 5′-CACATGGGCCTTTGGGGAATCTGTGCACTCCT-3′
Kbbα3S 5′-GCACAGATTCCCCAAAGGCCCATGTG-3′BamHI
AS 5′-GCGGGATCCTCACGCTAGAGAATGAGGCTC-3′
Site-directed mutagenesis
T18d-G197D5′-GCCAGACCTGAAGATGATGTCACCCTGAGG-3′
T18d-D198K5′-GCCAGACCTGAAGGTAAAGTCACCCTGAGG-3′
T18d-G197D/D198K5′-GCCAGACCTGAAGATAAAGTCACCCTGAGG-3′
T18d-T228M5′-TTGATTCAGGACATGGAGCTTGTC-3′
Kd-D197G/K198D5′-CAGCAGACCTGAAGGCGACGTCACCCTGAGGTGC-3′
Kb-M228T5′-GCTGATCCAGGACACGGAGCTCGTGGAGACCAGGCC-3′
  • a The positions of the mutated nucleotides in the primer DNA sequence are underlined. S, Sense; AS, antisense; RS, restriction site.