TABLE 1.
List of primers used for preparation of CacyBP/SIP mutants
| Mutant | Mutation | Template | Primers |
|---|---|---|---|
| CacyBP/SIP D11A/E14A | D11A | pET28a CacyBP/SIP | 5′-ttgcagaaagccctagaagaggtcaaagta-3′ |
| 5′-tactttgacctcttctagggctttctgcaa-3′ | |||
| E14A | pET28a CacyBP/SIP D11A | 5′-aaagccctagaagcggtcaaagtattgctg-3′ | |
| 5′-cagcaatactttgaccgcttctagggcttt-3′ | |||
| CacyBP/SIP D11H/E14K | D11H | pET28a CacyBP/SIP | 5′-caatactttgaccttttctaggtgtttctgcaactcttccaaaacggaagc-3′ |
| E14K | 5′-gcttccgttttggaagagttgcagaaacacctagaaaaggtcaaagtattg-3′ | ||
| CacyBP/SIP K25A/R26A | K25A | pET28a CacyBP/SIP | 5′-gtccactagggcaagactacgtgatac-3′ |
| 5′-gtatcacgtagtcttgccctagtggac-3′ | |||
| R26A | pET28a CacyBP/SIP K25A | 5′-gtacactagggcagcactacgtgatac-3′ | |
| 5′-gtatcacgtagtgctgccctagtggac-3′ | |||
| CacyBP/SIP K25E/R26E | K25E | pET28a CacyBP/SIP | 5′-ggaaaagtccactagggaagaactacgtgatactc-3′ |
| R26E | 5′-gagtatcacgtagttcttccctagacttttcc-3′ | ||
| CacyBP/SIP D11A/E14A/K25A/R26A | K25A | pET28a CacyBP/SIP D11A/E14A | 5′-caaagtattgctggaaaagtccactagggcagcactacgtgatactcttacaagtgaaaag-3′ |
| R26A | 5′-cttttcacttgtaagagtatcacgtagtgctgccctagtggacttttccagcaagcaatactttg-3′ | ||
| CacyBP/SIP S22D/T23D | S22D | pET28a CacyBP/SIP | 5′-agtattgctggaaaaggacactaggaaaagact-3′ |
| 5′-gtagtcttttcctagtgtccttttccagcaata-3′ | |||
| T23D | pET28a CacyBP/SIP S22D | 5′-agtattgctggaaaaggacgataggaaaagact-3′ | |
| 5′-gtagtcttttcctatcgtccttttccagcaata-3′ |
Substituted nucleotides are shown in boldface.