4.8. Mutagenesis of the PTTMP Promoter Sequence

The site directed mutagenesis of the promoter sequence was carried out by the overlap extension method using a Phusion DNA Polymerase in a GC buffer with 3 mM MgCl₂. The primers used to obtain the mutated promoter sequences are listed in Table 2. In the two-step PCR, the extension was 60 s at 60 °C temperature, and the other steps were performed according to the recommendations of the manufacturer, using the SensoQuest labcycler.