TABLE 1.
Sequences of primers used in the present study
| Name | Sequencea |
|---|---|
| RB-1 | 5′-ATATTAAGGATCCGGCGCCCATCACGG-3′ |
| RB-2 | 3′-ATTAATTAAGCTTCGTGACAACCTCCAGGT-5′ |
| RB-3 | 5′-CGCAATAGGATCCGGTGGACTTTATCCC-3′ |
| RB-4 | 5′-ATATTAGAATTCGCCAGCCCCCTG-3′ |
| RB-5 | 3′-TTATATAAGCTTGGGGGGGTCCTGGAG-5′ |
| RB-6 | 5′-ATCCTCGAATTCTGATGGGGGACA-3′ (mutant I) |
| RB-7 | 5′-GCATTAGAATTCCACCATGAATCA-3′ (mutant II) |
| RB-8 | 5′-AATTAAGAATTCGCCAGgggggTGATGGGGGCGACACTC-3′ (mutant IV) |
| RB-9 | 5′-AATTATGAATTCGCCAGgggggTGATcccccCGACACTC-3′ (mutant V) |
| RB-10 | 5′-TTAAATGAATTCGCCAGtttttTGATaaaaaCGACACTC-3′ (mutant VI) |
| RB-11 | 5′-ATATAAGAATTCGCCAGtttttTGATGGGG-3′ (mutant VIII) |
| RB-12 | 5′-ATATAAGAATTCGCCAGCCCCCTGATaaaaaCGACACTCCA-3′ (mutant IX) |
| RB-13 | 5′-ATATAAGAATTCGCCAGCCCCCTGATcccccCGACACTCCA-3′ (mutant IX-A) |
| RB-14 | 5′-ATATAAGAATTCGCCAGCCCCCcGgcGGGGGCGACACTCCA-3′ (mutant IX-B) |
| RB-15 | 5′-TTAATAGAATTCGCCAGCgCCCTGATGGGcGCGACACTC-3′ (mutant X) |
| RB-16 | 5′-TTAATAGAATTCGCCAGCCgCCTGATGGcGGCGACACTC-3′ (mutant XI) |
| RB-17 | 5′-TTAATAGAATTCGCCAGCCCgCTGATGcGGGCGACACTC-3′ (mutant XII) |
| RB-18 | 5′-ATATAAGAATTCGCCAGcccGGTGATCCgggCGACACTCC-3′ (mutant V-[A]) |
| RB-19 | 5′-ATATAAGAATTCGCCAGGcccGTGATCgggCCGACACTCC-3′ (mutant V-[B]) |
| RB-20 | 5′-ATATAAGAATTCGCCAGGGcccTGATgggCCCGACACTCC-3′ (mutant V-[C]) |
| RB-21 | 5′-TTAATAGAATTCGCCAGGcGGGTGATCCCgCCGACACTCC-3′ (mutant V-[D]) |
| RB-22 | 5′-TTAATAGAATTCGCCAGGccGGTGATCCggCCGACACTCC-3′ (mutant V-[E]) |
| RB-23 | 5′-TTAATAGAATTCGCCAGGcGcGTGATCgCgCCGACACTCC-3′ (mutant V-[F]) |
| RB-24 | 5′-ATTTATGAATTCGCCAGCtCCCTGATGGGaGCGACACTC-3′ (mutant XIX) |
| RB-25 | 5′-GTCTATGAATTCGCCAGCttCCTGATGGaaGCGACACTC-3′ (mutant XX) |
| RB-26 | 5′-GTCTATGAATTCGCCAGCtCtCTGATGaGaGCGACACTC-3′ (mutant XXI) |
| RB-27 | 3′-GCAACGAAGCTTCTCATACTAACGCC-5′ (mutant VII) |
| RB-28 | 5′-TATAATGAATTCCGCCGCCCACAGGAC-3′ |
| RB-29 | 3′-AATTTAAAGCTTTACCTCGAGGTTGCG-5′ |
| RB-30 | 5′-GCATATGAATTCGATGAACGGGGAGC-3′ |
| RB-31 | 3′-ATGGGCAAGCTTACATGATCTGCAGA-5′ |
| RB-32 | 3′-CGCGCCAAGCTTATTAAAGAAGGGAAAAAG-5′ |
| RB-33 | 5′-AGACCCAAGCTTAACGGGG-3′ |
| RB-34 | 3′-ACATGATCTGCAGAGAGGCC-5′ |
| RB-35 | 5′-TAATACGACTCACTATA AGACCCAAGCTTAACGGGGAGC-3′ |
| RB-36 | 3′-GGCTCACGGACCTTTCACAGCTA-5′ |
| RB-37 | 3′-CTAGGGCTAAGATGGAGCCACCA-5′ |
| RB-38 | 5′-CAGCGCAAGCTTTAGTAACCCTACCTC-3′ |
| RB-39 | 3′-CGCGGCGAGCTCCCTCCGAATTAAAGA-5′ |
a
The lowercase italic letters in the oligonucleotide sequences from RB-8 through RB-26 denote specific altered bases incorporated in the corresponding mutants. The boldface italic letters in forward primer sequence RB-35 represent the T7 polymerase promoter sequence fused to the primer. Restriction enzyme cleavage sites are underlined.