Table 5.

The oligonucleotide primers used in the PCR experiments for the construction of mutagenic CGTase DNA. The primers of EFP_BamHI and ERP_HindIII are forward and reverse external primers, respectively, and they contain restriction sequences for cloning purposes. The rest are the internal mutagenic primers containing the desired mismatched sequence.

Name of primer	Sequence *
EFP_BamHI	5′-CTCGGATCCGGACGTAACAAACAAAGTCAATTACTC-3′
ERP_HindIII	5′-GCCAAGCTTCCAATTAATCATAACCGTATCTGTTCCGG-3′
IFP_N28R	5′-CTGGCcgCAATCCTTCAGGCGCTATCTTTAG-3′
IRP_N28R	5′-AAGGATTGcgGCCAG GATTCCCGTCAGAGAATCG-3′
IFP_N132R	5′-CACGCCAcgcCATTCATCACCGGCACTTGAAACG-3′
IRP_N132R	5′-GATGAATGgcgTGGCGTGAAATCCATGATTACC-3′
IFP_S182G	5′-GATgGCATTTACAGAAACTTATATGATCTGGCAG-3′
IRP_S182G	5′-GTTTCTGTAAATGCcATCTTCATATGAAGAGAAATCTG-3′
IFP_S182E	5′-GAAGATgaaATTTACAGAAACTTATATGATCTGGCAG-3′
IRP_S182E	5′-TCTGTAAATttcATCTTCATATGAAGAGAAATCTG-3′

^*The restriction sequences are underlined and the mismatches are indicated in the bold lower-case letters.