Nucleic acid constructs and methods of making protein

The present invention relates to methods for producing a polypeptide, comprising: (a) cultivating a fungal host cell in a medium conducive for the production of the polypeptide, wherein the fungal host cell comprises a first polynucleotide comprising a nucleic acid sequence encoding the polypeptide operably linked to a copper-inducible promoter sequence comprising a copper-responsive upstream activation sequence activated by a copper-dependent trans-acting transcription factor and a second polynucleotide comprising one or more (several) additional copper-responsive upstream activation sequences operably linked upstream to the promoter sequence, wherein the promoter sequence is foreign to the nucleic acid sequence encoding the polypeptide and the copper-responsive upstream activation sequences are responsible for copper-induced transcription of the promoter sequence, and a third polynucleotide comprising at least one copy of a gene encoding the copper-dependent trans-acting transcription factor; and (b) isolating the polypeptide from the cultivation medium.