TABLE 1.
Primersa
| Construct | Mutation(s) | Primer direction | Primer sequence (5′–3′) |
|---|---|---|---|
| mα | Cys63 → Ala | F | GGGCGCTATgctGCTCTTGAGATGATA |
| R | TATCATCTCAAGAGCagcATAGCGCCC | ||
| mβ | Cys246 → Ala | F | CTTTGAGCATGGCCGCgcCTGGCTGAAGTTGTTC |
| R | GAACAACTTCAGCCAGgcGCGGCCATGCTCAAAG | ||
| mγ | Cys378 → Ala | F | CCTCACTGCTGGGTTCgcTTGGTTGCAGCTATTTCC |
| R | GGAAATAGCTGCAACCAAgcGAACCCAGCAGTGAGG | ||
| G164/G165 | Thr163 → Val/Gly164 → Val | F | CCATTCAACACGgtTGtAGGTGACGTTTATC |
| R | GATAAACGTCACCTaCAacCGTGTTGAATGG | ||
| G171/T172 | Leu170 → Ala/Gly171 → Ala | F | GGAGGTGACGTTTATCAGgcAGcAACCTGCACTATCGTTGAG |
| R | CTCAACGATAGTGCAGGTTgCTgcCTGATAAACGTCACCTCC | ||
| G271/V272 | Cys270 → Val/Gly271 → Val | F | CGGTTATCAATTGAACgtTGtCGTTCAAGGAAAATACATTGC |
| R | GCAATGTATTTTCCTTGAACGaCAacGTTCAATTGATAACCG | ||
| G350/G351 | Phe349 → Ala/Gly350 → Ala | F | TTAAGTTGCCAGGGAAGACTTATgcCGcTGGAAATGCCAGTTCGG |
| R | CCGAACTGGCATTTCCAgCGgcATAAGTCTTCCCTGGCAACTTAA | ||
| G484/G485 | Arg483 → Ala/Gly484 → Ala | F | CGTTACGGTCGCCGTgcgGcAGGCGGCAAGAAGTCAGGCCAATCG |
| R | CGATTGGCCTGACTTCTTGCCGCCTgCcgcACGGCGACCGTAACG | ||
| G485/G486 | Gly484 → Ala/Gly485 → Ala | F | CGTTACGGTCGCCGTCGAGcAGcCGGCAAGAAGTCAGGCCAATCG |
| R | CGATTGGCCTGACTTCTTGCCGgCTgCTCGACGGCGACCGTAACG |
a
F, forward; R, reverse. The substituted nucleotides in the primer sequences are indicated by lowercase letters.