Fig. 8
Schematic overview on the manually performed cloning. A Assembly of the new pBSYPGCW14Z expression plasmid by exchanging the PDC promoter of the commercially available pBSY3Z plasmid controlling the transcription of the GOI against the PGCW14 promoter. B Cloning strategy used for the assembly of the expression plasmids with the alternative SP by homologues recombination cloning. The pBSYPGCW14Z plasmid was used as backbone to insert the PCR-amplified yeast-originating OST1 or the artificial pre-OST1-pro-α-factor and a LPMO gene of which the natural SP had been removed (LPMO∆SP) by PCR