Constructs of glyceraldehyde-3-phosphate dehydrogenase promoter and methods for expressing genes using said constructs

- Chiron Corporation

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 desposited 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.

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Claims

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

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

4. A method for expressing and producing a heterologous protein in yeast, which comprises:

expressing a structural gene encoding said heterologous protein in a yeast expression vector having an inserted DNA construct to produce said heterologous protein, said inserted DNA construct comprising:
said structural gene being under the regulatory control of a first domain; and
said first domain being proximal to said structural gene, said first domain comprising at least about 200 bp and including the RNA polymerase binding site and transcription initiation site of the yeast Saccharomyces glyceraldehyde-3-phosphate dehydrogenase gene..Iaddend..Iadd.5. The method of claim 4 wherein said structural gene encodes Hepatitis B virus

surface antigen..Iaddend..Iadd.6. The method of claim 5 wherein said expression vector comprises the plasmid PHO5GAP1..Iaddend..Iadd.7. The method of claim 4 wherein said structural gene encodes a protein for vaccines..Iaddend..Iadd.8. The method of claim 4 wherein said first domain comprises about 200-500 bp..Iaddend..Iadd.9. The DNA construct of claim 1 wherein said first domain comprises about 200-500 bp..Iaddend.

Referenced Cited
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Other references
  • Van Vuuren et al. (1987) Am. J. Enol. Vitic. 38: 49-53, abstract cited. Bennetzen, J.L. et al., "Codon Selection in Yeast", J. Of Biol. Chem., 1982, 257(6), 3026-3031. Burnette, W.N., et al., Developments in Biological Standardization, "Properties and Relative Immunogenicity of Various Preparations of Recombinant DNA-Derived Hepatitis B Surface Antigen", (Congress Joint IABS/Who Symposium on Standardization and Control of Biologicals Produced by Recombinant DNA Technology), 1983, 59, 113-120. Burnette, W.N. et al., Modern Approaches to Vaccines, "Production of Hepatitis-B Recombinant Vacines", Cold Spring Harbor Laboratory, 1984, 245-250. Dobson, M.J. et al., "Conservation of high efficiency promoter sequences in Saccharomyces cerevisiae", Nucl. Acids Res., 1982, 10(8), 2625-2637. Highfield, P.E., Developments in Biological Standardization, "Impact of Genetic Engineering on Hepatitis B Virus", (Congress Joint IABS/Who Symposium on Standardization and Control of Biologicals Produced by Recombinant DNA Technology), 1983, 59, 121-124. Hitzeman, R.A. et al., "Expression of a human gene for interferon in yeast", Nature, 1981, 293, 717-722. Holland, M.J. et al., "Isolation and Characterization of a Gene Coding for Glyceraldehyde-3-phosphate Dehydrogenase from Saccharomyces cerevisiae", J. Biol. Chem., 1979, 254(12), 5466-5474. Holland, J.P. et al., "The Primary Structure of a Glyceraldehyde-3-phosphate Dehydrogenase Gene from Saccharomyces cerevisiae", J. Biol. Chem., 1979, 254(19), 9839-9845. Holland, J.P. et al., "Structural Comparison of Two Nontandemly Repeated Yeast Glyceraldehyde-3-Phosphate Dehydrogenase Genes", J. Biol Chem., 1980, 255(6), 2596-2605. Holland, J.J. et al., "Structure and Expression of Yeast Glycolytic Genes", 291-303. Miyanohara, A. et al., "Expression of hepatitis B surface antigen gene in yeast", Proc. Natl. Acad. Sci. USA, 1983, 80, 1-5. Musti, A.M. et al., "Transcriptional mapping of two yeast genes coding for glyceraldehyde 3-phosphate dehydrogenase isolated by sequence homology with wit chicken gene", Gene, 1983, 133-143. Oshima, T. et al., "Yeast Saccharomyces High Expression Vector", 5th Annual Meeting of the Japan Molecular Biology Institute (Tokyo), 1982, Lecture Synopses No. A1-34, 37. Tekamp-Olson, P. et al., "Regulatory Regions of Yeast Pyruvate Kinase and Glyceraldehyde-3-Phosphate Dehydrogenase Promoters", (Abstract), Meeting on the Molecular Biology of Yeast, 1993, 197. Tuite, M.F. et al., "Regulated high efficiency expression of human interferon-alpha in Saccharomyces cerevisiae", IRL Press Limited, Oxford, England, 1982, 603-608. Urdea, M.S. et al., "Chemical synthesis of a gene for human epidermal growth factor urogastrone and its expression in yeast", Proc. Natl. Acad. Sci. USA, 1983, 80, 7461-7465. Valenzuela, P. et al., "Characterization of Hepatitis B Surface Antigen Particles Produced in Yeast and in Mammalian Cells", (Abstract), Presented at the meeting on Modern Approaches to Vaccines, 1983, 70.
Patent History
Patent number: RE35749
Type: Grant
Filed: Sep 20, 1996
Date of Patent: Mar 17, 1998
Assignee: Chiron Corporation (Emeryville, CA)
Inventors: Steven Rosenberg (Oakland, CA), Patricia Tekamp-Olson (San Francisco, CA)
Primary Examiner: John L. LeGuyader
Attorneys: Francis A. Paintin, Barbara G. McClung, Robert P. Blackburn
Application Number: 8/710,744
Classifications
Current U.S. Class: 536/241; 435/693; 435/1723; 435/2542; 435/25421; 435/3201; 935/41
International Classification: C12P 1934; C12N 1511;