TABLE 1.
Primers and derived plasmids for site-directed mutagenesis
| Amino acid exchange | Primer sequence (5′-3′)a | Directionb | Resulting plasmid |
|---|---|---|---|
| C35S | GCACGCATTTCCGTTcgGGTGCGGTGGCGCTG | f | pCH1174 |
| CAGCGCCACCGCACCcgAACGGAAATGCGTGC | r | ||
| R63K | GCGATACGCTCGGGaaGCGCTTTGCGGTGAG | f | pCH1289 |
| CTCACCGCAAAGCGCttCCCGAGCGTATCGC | r | ||
| H70Q | GCGGTGAGCCTGCAgCCGCGGCTTGCCGCC | f | pCH1326 |
| GGCGGCAAGCCGCGGcTGCAGGCTCACCGC | r | ||
| R72K | TGAGCCTGCACCCGaaGCTTGCCGCCATCC | f | pCH1290 |
| GGATGGCGGCAAGCttCGGGTGCAGGCTCA | r | ||
| C84S | CGGCGCGACGTCACCTcgTTCCATCACGACAGC | f | pCH1176 |
| CTGTCGTGATGGAAcgAGGTGACGTCGCGCCGC | r | ||
| F85L | GACGTCACCTGCcTgCATCACGACAGCATG | f | pCH1291 |
| CATGCTGTCGTGATGcAgGCAGGTGACGTC | r | ||
| D93N | ACAGCATGTTGCCCaACCCCTACGACGGGC | f | pCH1292 |
| GCCCGTCGTAGGGGTtGGGCAACATGCTG | r | ||
| Y95F | GTTGCCCGACCCCTtCGACGGGCTGATCG | f | pCH1149 |
| CGATCAGCCCGTCGaAGGGGTCGGGCAAC | r | ||
| Y95L | GTTGCCCGACCCCctgGACGGGCTGATCG | f | pCH1150 |
| CGATCAGCCCGTCcagGGGGTCGGGCAAC | r | ||
| D96N | TGCCCGACCCCTACaACGGGCTGATCGACG | f | pCH1293 |
| CGTCGATCAGCCCGTtGTAGGGGTCGGGC | r | ||
| H110Q | GCCGCTGCCGGTGCAgGACTGCATGGGCACG | f | pCH1151 |
| CGTGCCCATGCAGTCcTGCACCGGCAGCGGC | r | ||
| C112S | CTGCCGGTGCACGACTcgATGGGCACGAGCCTGC | f | pCH1152 |
| GCAGGCTCGTGCCCATcgAGTCGTGCACCGGCAG | r | ||
| D130N | GCGTGCTGACGCTCaACGCGCTCACGGTCGG | f | pCH1325 |
| CCGACCGTGAGCGCGTtGAGCGTCAGCACGC | r | ||
| F137L | TCACGGTCGGCACGcTgGACGCCGCCGCGC | f | pCH1327 |
| GCGCGGCGGCGTCcAgCGTGCCGACCGTG | r |
a
Mutated bases are indicated in lowercase.
b
f, forward primer; r, reverse primer.