... vector to an EcoRV site. The mutagenesis oligonucleo-tides were 5¢-GTAACTGTAAGAGAACTGGTCAC-3¢(Lys70 to Arg), 5¢-GTAACTGTAGCAGAACTGGTCAG-3¢ (Lys70 to Ala), and 5¢-GGTAACTGTATATGAACTGGTCAG-3¢ ... Inc., Palo Alto, CA, USA) using the method ofDeng and Nicloff [19], as directed by the manufacturer. Theselection oligonucleotide, 5¢-AAATGCTTCAATGATATCGAAAAAGGAAG-3¢, converted a unique SspIsiteonthe ... europaea, cyp,was amplified by PCR from a 7.8-kb BamHI fragment ofgenomic DNA which had previously been subcloned intothe plasmid vector pUC119 [16]. The forward primer,5¢-GCTACCATATGAAAACAGCTTGGTAGGT-3¢,en-compassed...