New fungal orotic acid phosphoribosyltransferase gene, useful as a selection marker in recombinant protein expression systems

摘要

A DNA sequence (I) comprising: (1) a 1133-1877 nucleotide (nt) region of a 3448 nt sequence (S1) fully defined in the specification; (2) a 648 nt sequence (S2); (3) a sequence 60% homologous with (S1) or (S2); or (4) encodes a protein with orotic acid phosphoribosyltransferase activity (pyrF) activity, is new. Independent claims are also included for the following: (1) a protein (II) with pyrF activity that has a 227 amino acid (aa) sequence (S3) fully defined in the specification, or is over 60% homologous with (S3); (2) an expression vector (III) containing (I); (3) microorganisms containing (III); (4) production of fungal strains (A), capable of efficient expression and secretion of proteins, by transforming a uridine-auxotrophic strain (IV) of Trametes, Coriolus or Polysporus having a defect in the pyrF gene; (5) an expression system (V) containing (IV) and an expression vector carrying a selection marker and a pyrF gene of Basidiomycetes; and (6) producing proteins using (V) or microorganisms containing (III).