TABLE 4.
List of oligonucleotide primers used to generate the blaPER-2 allelic variants and recombinant plasmids encoding the Arg220 and Thr237 PER-2 mutants
| Primer namea | Primer sequence (5′→3′)b | Recombinant plasmid |
|---|---|---|
| PER2-RxL-Fw | GAAACCACCACAGGGCCCCAGCTGTTAAAAGGCTTGTTACCTGC | pK19/PER-2 Arg220Leu |
| PER2-RxL-Rv | GCAGGTAACAAGCCTTTTAACAGCTGGGGCCCTGTGGTGGTTTC | |
| PER2-RxS-Fw | GAAACCACCACAGGGCCCCAGAGCTTAAAAGGCTTGTTACCTGC | pK19/PER-2 Arg220Ser |
| PER2-RxS-Rv | GCAGGTAACAAGCCTTTTAAGCTCTGGGGCCCTGTGGTGGTTTC | |
| PER2-RxG-Fw | GAAACCACCACAGGGCCCCAGGGCTTAAAAGGCTTGTTACCTGC | pK19/PER-2 Arg220Gly |
| PER2-RxG-Rv | GCAGGTAACAAGCCTTTTAAGCCCTGGGGCCCTGTGGTGGTTTC | |
| PER2-RxH-Fw | GAAACCACCACAGGGCCCCAGCATTTAAAAGGCTTGTTACCTGC | pK19/PER-2 Arg220His |
| PER2-RxH-Rv | GCAGGTAACAAGCCTTTTAAATGCTGGGGCCCTGTGGTGGTTTC | |
| PER2-RxT-Fw | GAAACCACCACAGGGCCCCAGACGTTAAAAGGCTTGTTACCTGC | pK19/PER-2 Arg220Thr |
| PER2-RxT-Rv | GCAGGTAACAAGCCTTTTAACGTCTGGGGCCCTGTGGTGGTTTC | |
| PER2-RxC-Fw | GAAACCACCACAGGGCCCCAGTGCTTAAAAGGCTTGTTACCTGC | pK19/PER-2 Arg220Cys |
| PER2-RxC-Rv | GCAGGTAACAAGCCTTTTAAGCACTGGGGCCCTGTGGTGGTTTC | |
| PER2-T237A-Fw | TAAAACCGGTGCTTCGGGCGTCAGAGCAGGAAAAACTGC | pK19/PER-2 Thr237Ala |
| PER2-T237A-Rv | CTCTGACGCCCGAAGCACCGGTTTTATGCGCCACTATA |
a
Fw, forward primer; Rv, reverse primer.
b
Nucleotides that differ from the original sequence are shown in boldface. Restriction sites for the ApaI enzyme generated by a silent mutation are underlined.