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

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 α 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.

What is claimed is: 
     
       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.!. .Iadd.said .Iaddend.first domain .Iadd.being .Iaddend.proximal to said structural gene, said first domain comprising .Iadd.at least .Iaddend.about 200. .-500.!. bp, and including the RNA polymerase binding site and transcription initiation site of the yeast Saccharomyces glyceraldehyde-3-phosphate dehydrogenase gene.   
     
     
       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.