Patent ID: 6783985
Filing Date: 2004-08-31
Classification: A61K,C07K,C12N,C12R

Abstract:
A method for constructing a strain of diploid fungal cells in which both alleles of a gene are modified, the method comprising the steps of:(a) modifying a first allele of a gene in diploid fungal cells by recombination using a gene disruption cassette comprising a first nucleotide sequence encoding a first expressible selectable marker, thereby providing heterozygous diploid fungal cells in which the first allele of the gene is inactivated; and (b) modifying the second allele of the gene in the diploid fungal cells by recombination using a promoter replacement fragment comprising a second nucleotide sequence encoding a second expressible selectable marker and a heterologous promoter, such that expression of the second allele of the gene is regulated by the heterologous promoter wherein at least one of the first and the second expressible selectable marker is a drug-resistance marker, thereby constructing a strain of diploid fungal cells in which both alleles of a gene are modified.