High copy number recombinant expression construct for regulated high-level production of polypeptides in yeast

摘要

The invention provides methods and recombinant expression constructs for inducing and sustaining high-level production of a recombinant polypeptide in yeast. The invention specifically provides high copy number recombinant expression constructs that express high levels of trans-acting transcription factors that in turn induce expression of a recombinant nucleic acid encoding a heterologous or endogenous recombinant polypeptide. The invention more specifically provides constructs that express galactose-inducible and temperature-sensitive transcription factors. The invention also provides constructs comprising nucleic acids the transcription of which is regulated by the transcription factors expressed by the construct. The invention also provides yeast cells transformed by the recombinant expression constructs of the invention that permit sustained high-level expression of a recombinant polypeptide. The invention further provides methods for producing yeast cells transformed with the recombinant expression constructs of the invention, and methods for inducing high-level expression of a recombinant polypeptide in said transformed yeast cells. The invention also provides such recombinant polypeptides produced by the methods and the transformed yeast cells of the invention.