Table 1 Primers for site-directed mutagenesis
Primers | Primer sequences (5'-3') | Tm (°C) |
|---|---|---|
S76M-F | 5’- CCACCGTAatgAATGGTAAGTTGAAGTTAACAACTAAGCG − 3’ | 63.1 |
S76M-R | 5’- ACCATTcatTACGGTGGCATTTTCATTACGC − 3’ | 62.7 |
G96M-F | 5’- GGGATatgTGTAATCAGCAGCAGGTAGCAAA − 3’ | 60.6 |
G96M-R | 5’- CTGATTACAcatATCCCAAAATGTACGATTATGTGTTT − 3’ | 60.7 |
C97K-F | 5’- GGATGGCaaaAATCAGCAGCAGGTAGCAAATTATC − 3’ | 61.6 |
C97K-R | 5’- GCTGATTtttGCCATCCCAAAATGTACGATTAT − 3’ | 60.2 |
Q100M-F | 5’- AATCAGatgCAGGTAGCAAATTATCCTCTGTACTATACA − 3’ | 61.3 |
Q100M-R | 5’- GCTACCTGcatCTGATTACAGCCATCCCAAAATG − 3’ | 61.4 |
T151S-F | 5’- AGATCGCagcTTAACTGAAAATGGTGATGTACAGTACAG − 3’ | 61.0 |
T151S-R | 5’- CAGTTAAgctGCGATCTATTGTGCTATACATCCAA − 3’ | 60.8 |
L152R-F | 5’- TCGCACAcgtACTGAAAATGGTGATGTACAGTACAGTG − 3’ | 61.2 |
L152R-R | 5’- TTTCAGTacgTGTGCGATCTATTGTGCTATACATCC − 3’ | 62.7 |
T153L-R | 5’- GATCGCACATTActgGAAAATGGTGATGTACAGTACAGTGAAA-3’ | 61.3 |
T153L-F | 5’- TCcagTAATGTGCGATCTATTGTGCTATACAT-3’ | 60.1 |
E154R-F | 5’- AACTcgtAATGGTGATGTACAGTACAGTGAAATTG − 3’ | 61.2 |
E154R-R | 5’- CATCACCATTacgAGTTAATGTGCGATCTATTGTGCTATAC − 3’ | 60.5 |
N155L-F | 5’- AACTGAActgGGTGATGTACAGTACAGTGAAATTGATG − 3’ | 61.4 |
N155L-R | 5’- CATCACCcagTTCAGTTAATGTGCGATCTATTGTGC − 3’ | 62.9 |
S190R-F | 5’- AAATGGCcgtCCTACTTGGATGCGACCGG − 3’ | 61.1 |
S190R-R | 5’- AAGTAGGacgGCCATTTTTAACGACAATGTTATGT − 3’ | 60.0 |
T192R-F | 5’- AATGGCTCTCCTcgtTGGATGCGACCGGGTAGTG − 3’ | 62.2 |
T192R-R | 5’- CAacgAGGAGAGCCATTTTTAACGACAA − 3’ | 60.3 |
G197M-F | 5’- TTGGATGCGACCGatgAGTGCGCCAGAAACAAATCATA − 3’ | 60.2 |
G197M-R | 5’- TcatCGGTCGCATCCAAGTAGGAG − 3’ | 60.5 |
E201P-F | 5’- CGCCAccgACAAATCATAATGGCTATCACTTACCG − 3’ | 62.6 |
E201P-R | 5’- ATGATTTGTcggTGGCGCACTACCCGGTCG − 3’ | 65.7 |
T202M-F | 5’- GCGCCAGAAatgAATCATAATGGCTATCACTTACCGTT − 3’ | 60.3 |
T202M-R | 5’- TGATTcatTTCTGGCGCACTACCCGG − 3’ | 62.5 |
E279D-F | 5’- ATCAGCAgatGGCTTTCCTACCTCAATGGAAGT − 3’ | 60.9 |
E279D-R | 5’- GAAAGCCatcTGCTGATTTATTAGCAGATGGGTAA − 3’ | 60.6 |