TABLE 4.
Degenerate primers used for site-directed saturation mutagenesis of the ChPPK
| Primer | Oligonucleotide sequences (5′−3′) a , b |
|---|---|
| A79-S-F | TGGATGCANNKGGTAAAGATGGTAC |
| A79-S-R | TCTTTACCMNNTGCATCCATTGCC |
| G80-S-F | ATGCAGCANNKAAAGATGGTACCGTT |
| G80-S-R | ACCATCTTTMNNTGCTGCATCCATTG |
| K81-S-F | AGCAGGTNNKGATGGTACCGTTAAACATAT |
| K81-S-R | TACCATCMNNACCTGCTGCATCCATTG |
| F102-S-F | TGACCAGCNNKAAAGTTCCGTCCAAAAT |
| F102-S-R | ACGGAACTTT MNNGCTGGTCACTTTAACA |
| K103-S-F | ACCAGCTTTNNKGTTCCGTCCAAAATT |
| K103-S-R | ACGGAACMNNAAAGCTGGTCACTTT |
| P105-S-F | TTTAAAGTTNNKTCCAAAATTGAACTGAG |
| P105-S-R | ATTTTGGAMNNAACTTTAAAGCTGG |
| S106-S-F | AGTTCCGNNKAAAATTGAACTGAGT |
| S106-S-R | TCAATTTTMNNCGGAACTTTAAAGCTGG |
| I108-S-F | CGTCCAAANNKGAACTGAGTCATGA |
| I108-S-R | TCAGTTCMNNTTTGGACGGAACTTTAAAG |
| S111-S-F | TTGAACTGNNKCATGATTATCTGTGGC |
| S111-S-R | TAATCATGMNNCAGTTCAATTTTGGACG |
| R117-S-F | TATCTGTGGNNKCATTATGTGGCAC |
| R117-S-R | ACATAATGMNNCCACAGATAATCATGA |
| L285-S-F | ACCGTGAGCNNKGAACAGAAAGCGG |
| L285-S-R | TTCTGTTCMNNGCTCACGGTCGGAAA |
a
Bold-marked bases in the primers indicate positions that contain the degenerate codon.
b
N = A, T, G, C (equimolar amounts); K = G, T (equimolar amounts); M = A, C (equimolar amounts).