Transformation of Penicillium chrysogenum using dominant selection markers and expression of an Escherichia coli lacZ fusion gene.
An industrial Penicillium chrysogenum strain was transformed using two dominant selection markers, namely the bacterial gene for phleomycin resistance (ble) fused to a fungal promoter, and the acetamidase (amdS) gene from Aspergillus nidulans. Transformation frequencies of up to 20 transformants per microgram of DNA were obtained with the ble system. With the amdS marker the frequency was up to 120 transformants. Cotransformation was very efficient when using amdS as a selection marker. The introduction of pAN5-41B, a plasmid carrying the Escherichia coli lacZ gene fused to the strong glyceraldehyde-3-phosphate dehydrogenase gene (gpd) promoter from A. nidulans, resulted in the formation of blue colonies on XGal plates indicating expression of the lacZ fusion gene in P. chrysogenum. A more detailed analysis of expression levels in several transformants showed that up to 6% of the total amount of soluble protein consists of the beta-galactosidase fusion protein.
D000581:Amidohydrolases / Q000096:biosynthesis / Q000235:genetics; D001233:Aspergillus nidulans / Q000235:genetics; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics; D010407:Penicillium / Q000235:genetics*; D010408:Penicillium chrysogenum / Q000187:drug effects / Q000235:genetics*; D010692:Phleomycins / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics; D014170:Transformation, Genetic; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D010692:Phleomycins; D011993:Recombinant Fusion Proteins; D001616:beta-Galactosidase; D000581:Amidohydrolases; C021445:acetamidase