Homologous transformation of the lignin-degrading basidiomycete Phanerochaete chrysosporium

Summary A clone containing the Phanerochaete chrysosporium ade1 gene was isolated from a EMBL3 genomic library using the ade5 gene encoding aminoimidazole ribonucleotide synthetase, from Schizophyllum commune, as a probe. A 6.0 kb fragment incorporating the ade1 gene was subcloned into pUC18 (pADE1) and used to transform the P. chrysosporium ade1 auxotrophic strain. Transformation frequencies were similar to those obtained previously with the S. commune ade5 gene; however, homologous transformants arose earlier than heterologous transformants. The transformants were mitotically and meiotically stable and Southern blot analysis indicated that the plasmid, pADE1, integrated ectopically in single or multiple copies. The pADE1 insert was mapped for restriction sites and the approximate location of the ade1 gene within the insert was determined.