# US 5,349,059 A · Hybrid promoter contructs of glyceraldehyde-3-phosphate dehydrogenase promoter

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- Number: US 5,349,059 A
- Document ID: US5349059A
- Status: Granted
- Kind: A
- Application number: 08/042,134
- Filed: 1993-04-02
- Granted: 1994-09-20
- Earliest claimed: 1983-02-22
- Assignees: Chiron Corporation
- Inventors: Steven Rosenberg; Patricia Tekamp-Olson
- Web page: https://patentgrep.com/patent/US5349059A

## CPC

- C07K 14/005: from viruses
- C07K 14/8125: {Alpha-1-antitrypsin}
- C12N 9/0089: {acting on superoxide as acceptor (1.15)}
- C12N 15/62: DNA sequences coding for fusion proteins
- C12N 15/81: for yeasts
- C07K 2319/00: Fusion polypeptide
- C07K 2319/02: containing a signal sequence
- C07K 2319/40: containing a tag for immunodetection, or an epitope for immunisation
- C07K 2319/75: containing a fusion for activation of a cell surface receptor, e.g. thrombopoeitin, NPY and other peptide hormones
- C12N 2730/10122: New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Yeast promoters of glycolytic enzymes are modified by isolating a fragment encompassing the RNA polymerase binding site and joining to the 5' end of this fragment a DNA sequence providing for enhanced inducible or constitutive transcription of a structural gene. Constructs are prepared for efficient expression of foreign genes in yeast. Yeast strains 2150-2-3(pC1/1GAPSOD) and AB110(pC1/1GAPATi9), producing human .alpha..sub.1 -antitrypsin and superoxide dismutase, were deposited at the A.T.C.C. on May 9, 1984 and given Accession Nos. 20708 and 20709, respectively; and 2150-2-3(GAP5), 2150-2-3(Pyk5) and 2150-2-3(PHO5GAP1), expressing Hepatitis B surface antigen, were deposited at the A.T.C.C. on May 9, 1984 and given Accession Nos. 20705, 20706 and 20707, respectively.

## Claims

### Claim 1

A DNA construct for expression of heterologous proteins, wherein said construct comprises:
a structural gene encoding said heterologous protein, wherein said structural gene is under the regulatory control of a first domain; and
a first domain proximal to said structural gene, said first domain comprising about 200-500 bp, and including the RNA polymerase binding site and transcription initiation site of the yeast Saccharomyces glyceraldehyde-3-phosphate dehydrogenase gene.

### Claim 2 (depends on 1)

The DNA construct of claim 1, wherein said structural gene encodes Hepatitis B virus surface antigen.

### Claim 3 (depends on 2)

The DNA construct of claim 2, wherein said construct is plasmid PHO5GAP1.