Light-regulated promoters for production of heterologous proteins in filamentous fungi

• US 5,741,665 A
• Filed: 05/10/1994
• Issued: 04/21/1998
• Est. Priority Date: 05/10/1994
• Status: Expired due to Term

• Current Assignee: University of Hawai'i
• Original Assignee: University of Hawai'i
• Inventors: Stuart, W. Dorsey, Kato, Elie K.
• Primary Examiner(s): Chambers, Jasemine C.
• Assistant Examiner(s): PRIEBE, SCOTT DAVID
• Application Number:
  US08/240,372
• Time in Patent Office: 1,442 Days
• Field of Search: 435/69.1, 435/254.4, 536/24.1
• US Class Current: 435/69.1
• CPC Class Codes: C12N 15/80 for fungi
• CPC Class Codes: C12N 9/6481 Pepsins (3.4.23.1; 3.4.23.2; 3.4.23.3)

Abstract

The al-3 and related promoters can be used to provide light-regulated recombinant production of heterologous proteins in filamentous fungi. Expression systems utilizing these promoters can be placed in vectors which also optionally contain selectable marker means.

Claims

1. A nucleic acid molecule for expression of a first nucleotide sequence encoding a heterologous protein in a filamentous fungus, which nucleic acid molecule comprises said first nucleotide sequence operably linked to the al-1, al-2 or al-3 promoter and optionally further comprises a second nucleotide sequence that provides for a selectable marker in said fungus.

2. The nucleic acid molecule of claim 1 wherein said second nucleotide sequence is present and promotes homologous recombination of said nucleic acid molecule with the fungus chromosome so as to alter a region of said chromosome thus providing a selectable marker.

3. The nucleic acid molecule of claim 2 wherein said second sequence is contiguous with said first sequence.

4. The nucleic acid molecule of claim 2 wherein said second sequence corresponds to at least a portion of the mtr locus.

5. The nucleic acid molecule of claim 1 wherein said second sequence is present and encodes a protein that confers a selectable characteristic on said fungus.

6. The nucleic acid molecule of claim 5 wherein said selectable characteristic is conferred by the product of the mtr gene.

7. The nucleic acid molecule of claim 1 wherein said first sequence encodes chymosin or relaxin.

8. A Neurospora fungus modified to contain the nucleic acid molecule of claim 1.

9. A method to produce proteins heterologous to a Neurospora fungus, which method comprises culturing the Neurospora fungus of claim 8 under conditions wherein said heterologous protein is produced;

and

recovering the heterologous protein from the culture.