Efficient transformation of monokaryons from Pleurotus salmoneostramineus using the GPD promotor for hygromycin B resistance

Abstract

A newly isolated glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter from Pleurotus salmoneostramineus was used for the expression of a hygromycin B resistance gene in a transformation study of the monokaryons, which are monosporous isolates from P. salmoneostramineus NBRC31859. The promoter site was set at 1,100 bp upstream of this gene, the hygromycin B resistance gene was introduced, and colonies resistant to 50 mg/mL hygromycin B were detected. Southern blotting analysis of transformants indicated that the integration of the gene occurred by non-homologous end-joining. These methods could be useful for functional gene analyses to study monokaryotic fruit body formation in P. salmoneostramineus.