Fungus wherein the areA gene has been modified and an areA gene from Aspergillus oryzae

• US 6,025,185 A
• Filed: 12/03/1996
• Issued: 02/15/2000
• Est. Priority Date: 06/17/1994
• Status: Expired due to Term

• Current Assignee: Novozymes A/S (Novozymes)
• Original Assignee: NOVO Nordisk Biotech Incorporated (Novo Nordisk A/S)
• Inventors: Hynes, Michael J., Christensen, Tove
• Primary Examiner(s): Kemmerer, Elizabeth
• Assistant Examiner(s): Romeo, David S.
• Application Number:
  US08/750,458
• Time in Patent Office: 1,169 Days
• Field of Search: 435/69.1, 435/253.2, 435/325, 435/254.11, 435/172.1, 435/91.4, 536/23.74
• US Class Current: 435/254.11
• CPC Class Codes: C12N 15/01 Preparation of mutants without inserting foreign genetic material therein; Screening processes therefor
• CPC Class Codes: C12N 9/62 from Aspergillus
• CPC Class Codes: C12N 9/6427 Chymotrypsins (3.4.21.1; 3.4.21.2); Trypsin (3.4.21.4)
• CPC Class Codes: C12N 9/6481 Pepsins (3.4.23.1; 3.4.23.2; 3.4.23.3)

Abstract

A modified Aspergillus oryzae cell having an area gene which does not express a functional AreA activator, DNA sequences encoding the area gene, and method for using the cell of the invention for production of a heterologous polypeptide.

Claims

1. An Aspergillus oryzae cell comprising a modified areA gene, wherein said modification reduces the synthesis of a functional areA gene product and wherein the modification of the areA gene is

(a) a deletion of all or parts of SEQ ID NO:

1;

or

(b) insertion of extra DNA internally into SEQ ID NO;

1.

2. A method for producing the Aspergillus oryzae cell of claim 1, wherein said method comprises:

(a) cloning the area gene comprising SEQ ID NO;

1 from Aspergillus oryzae;

(b) producing a DNA construct comprising the area gene wherein an internal part of SEQ ID NO;

1 has been substituted or deleted, or extra DNA has been inserted into SEQ ID NO;

1;

(c) transforming said Aspergillus oryzae with the construct; and

(d) selecting transformants which exhibit a reduced synthesis of a functional area gene product.

3. A process for producing a polypeptide comprising the steps of:

(a) cultivating the Aspergillus oryzae cell of claim 1; and

(b) recovering and purifying the polypeptide.

4. The process of claim 3, wherein the Aspergillus oryzae cell has been transformed with a DNA sequence coding for the polypeptide.

5. The process of claim 3, wherein the polypeptide is a fungal polypeptide.

6. The process of claim 3, wherein the polypeptide is a secreted protein.

7. The process of claim 6, wherein the polypeptide is secreted to the extracellular medium.

8. An isolated DNA sequence coding for the areA gene from Aspergillus oryzae comprising SEQ ID NO:

1.