Table 1.
Sets of mismatched primers used to create the M1, M2, and DM mutants of SMEZ-2
| Mutation | Primer sequence |
|---|---|
| W75La,b | 5′-CCATTTGATTTGAACTATTTATC-3′ |
| 5′-GATAAATAGTTCAAATCAAATGG-3′ | |
| K182Qa,b | 5′-GATATAGAGATCAAGAAAGTATC-3′ |
| 5′-GATACTTTCTTGATCTCTATATC-3′ | |
| D42C1a,b,c | 5′-GATGTTAGATGTGCTAGAGATTTC-3′ |
| 5′-CTCTAGCACATCTAACATCAAGTTTC-3′ | |
| Y18Ab,c | 5′-CGATTGTAGCTGAATATTCAGATATAG-3′ |
| 5′-GAATATTCAGCTACAATCGTACTATAG-3′ | |
| H202A.D204Ac | 5′-GGGGCTTTAGCTATAGAAATTGACTCC-3′ |
| 5′-AGCTAAAGCCCCAATTTTATCTATATTG-3′ |
a
SMEZ-2 M1 mutant was created by mutating 3 TCR-binding residues (W57L, K182Q, D42C). This mutant bound to MHC class II but not the TCR.
b
SMEZ-2 M2 mutant was created by mutating 4 TCR-binding residues (Y18A, W75L, K182Q, D42C). This mutant bound to MHC class II but not the TCR.
c
SMEZ-2 DM mutant was created by mutating two TCR-binding residues (Y18A, D42C) and two MHC class II-binding residues (H202A, D204A). This mutant no longer bound to MHC class II or the TCR.