Table 1.
Primers used in this study1.
| Cloning of genomic tcdE | |
| SpeI-tcdE-F | ACTAGTATGCACAGTAGTTCACC |
| BamHI-tcdE-R | GGATCCCTTTTCACCCTTAGCATTC |
| Cloning of tcdE and its variants | |
| tcdE-M1-NdeI-F | ATTATCATATGCACAGTAGTTCACCTTTTTATATTTC |
| tcdE-BamHI-R | ATATTGGATCCCTTTTCATCCTTAGCATTCATTTC |
| tcdE-S64-BamHI-R | TATATGGATCCAGAATTAAATTTACGACTTTTTATTGC |
| tcdE-I104-BamHI-R | TATATGGATCCGATACAATCTTGTGGTAACATAAATAAAAAG |
| tcdE-P142-BamHI-R | TATATGGATCCAGGTACTGGTAATCCACATAAGCAC |
| tcdE-Met25-NdeI-F | ATTATCATATGAATATGACAATATCTTTTTTATCAG |
| tcdE-Met27-NdeI-F | ATTATCATATGACAATATCTTTTTTATCATGAGC |
| Cloning of tcdL | |
| tcdL-synth-NdeI-F | ATTATCATATGCCAAGAGACACACAAGTATTAAATACATATAATTTCGAAGCAAGTGTTCATTACTATATGGATGACAAGGTAGTATATC |
| tcdL-synth-BamHI-R | ATATTGGATCCATAGATTTTACCAACTGACCATGCACCATCTTTGTGAACCAATGTTTGATATACTACCTTGTCATCCATATAGTAATG |
| tcdL-NdeI-F | ATTATCATATGCCAAGAGACACACAAG |
| tcdL-BamHI-F | ATATTGGATCCATAGATTTTACCAACTG |
| Cloning of lepB | |
| NdeI-lepB-F | TATTACATATGGCGAATATGTTTGCCCTGATTCTGG |
| BamHI-lepB-R | TAATAGGATCCATGGATGCCGCCAATGCGACTTAAGC |
| Single amino acid exchanges2 | |
| tcdEM25/27L-F | AACTTTATAAATATATGCTCTGATAAAAAAGATATTGTGAGATTGAGAACGCCTCCTAGGTTTATATAAAAAAATATTTTGTTACCATTAG |
| tcdEΔRBS-F | GCTCTGATAAAAAAGATATTGTCATATTCATAACGAGTAATAGGTTTATATAAAAAAATATTTTGTTACCATTAGAAATATAAAAAGGTGA |
| tcdE-M25M27L-F | ATATGCTCTGATAAAAAAGATATTGTGAGATTCATATGTATATCTCCTTCTTAAGAATTGTTC |
| tcdE-I151>V-F | GATTAAAGGAAAAAATAGCAGTTTTACTAGATGCAATGACAG |
| tcdB-sil-NdeI-F | CTTAAGTGGCCCTGAAGCGTATGCGGCAGCTTATCAAG |
| Bacterial-2-hybrid screen | |
| 2H-tcdE-BamHI-F | TATATGGATCCCACAGTAGTTCACCTTTTTATATTTC |
| 2H-tcdE-KpnI-R | TATATGGTACCCCCTTTTCATCCTTAGCATTC |
| 2H-tcdB-BamHI-F | TATATGGATCCGATAAACTTGTTCACTTAAATC |
| 2H-tcdB-KpnI-R | TATGGATCCAGTTTAGTTAATAGAAAACAGTTAG |
| 2H-tcdL-BamHI-F | TATATGGATCCCCAAGAGACACACAAGTATTAAATAC |
| 2H-tcdL-KpnI-R | TATATGGTACCCCATAGATTTTACCAACTGAC |
1Primer sequences are given in 5′-3′ direction. 2Corresponding reverse primers covered the same sequence.