Chiron Corporation

Jeffrey R Shuster

Patricia A Tekamp-Olson

Steven Rosenberg

Pablo D Valenzuela

Rae Lyn Burke

Lawrence S Cousens

James P Merryweather

Philip J Barr

A method is provided for expressing non-yeast proteins under the control of ADH2 regulatory sequences in a yeast host having enhanced expression of ADR I. Transformed yeast hosts are also provided.

High level expression of proteins in yeast

Efficient integration of heterologous DNA into yeast genomic DNA is accomplished at high copy number by targeting integration vectors to dispersed repetitive elements such as DELTA sequences, Ty elements, or tRNA DNA sequences present in the host cell genome.

Method for integrative transformation of yeast using dispersed

Novel methods and compositions are provided for enhanced yield of heterologous proteins in fungi. The method and compositions involve employing fusion sequences involving a sequence encoding a heterologous product produced in relatively large amount as a stable polypeptide in the host fused to a second sequence in open reading frame with the prior sequence coding for a different heterologous polypeptide, where the two polypeptides are joined by a selectively cleavable linkage. In particular, a sequence coding for superoxide dismutase is joined to another polypeptide of interest at either terminus of the superoxide dismutase in a yeast expression vector under transcriptional control of an active promoter and the vector introduced into a yeast host and the host grown. High yields of the fusion product are obtained in this manner, where the fusion product can be selectively cleaved so as to produce both the superoxide dismutase and the other polypeptide in high yield. The S. cerevisiae strain 2150-2-3 (pYASI1) was deposited at the A.T.C.C. on Feb. 27, 1985 and given accession no. 20745. The S. cerevisiae strain AB110 (pYLUIGF2-14) was deposited at the A.T.C.C. on Mar. 19, 1986 and given accession no. 20796.

Superoxide dismutase fusion polypeptides for expression of mammalian

Methods and compositions are provided for efficient expression of genes in unicellular microoorganisms, particularly yeast. The systems involve an expression system employing transcriptional initiation regions from glycolytic enzymes, particularly a chimeric expression system, having a first region providing for regulatable or constitutive expression, a second region providing for transcriptional initiation, where regions one and two are not found joined together in functional relationship in nature, and optionally a sequence providing for a secretory leader and processing signal, where the expression cassette will be joined to a gene which may be homologous or heterologous to the host. The expression cassette can be used on an extrachromosomal element or integrated into the host genome, whereby continuous expression can be achieved or inducible expression is obtained, by virtue of the presence or absence of an inducer. Constructions may be provided, where structural genes may be introduced in reading frame with the cassette to provide for expression of the natural peptide, a fused peptide, or a peptide precursor. The S. cerevisiae strain AB110 (pAB24/P. vivax 1-5 was deposited at the A.T.C.C. on May 9, 1986, and given Accession No. 20797.

Eukaryotic regulatable transcription

Methods and compositions are provided for efficient expression of genes in unicellular microorganisms, particularly yeast. The systems involve an expression system employing transcriptional initiation regions from glycolytic enzymes, particularly a chimeric expression system, having a first region providing for regulatable or constitutive expression, a second region providing for transcriptional initiation, where regions one and two are not found joined together in functional relationship in nature, and optionally a sequence providing for a secretory leader and processing signal, where the expression cassette will be joined to a gene which may be homologous or heterologous to the host. The expression cassette can be used on an extrachromosomal element or integrated into the host genome, whereby continuous expression can be achieved or inducible expression is obtained, by virtue of the presence or absence of an inducer. Constructions may be provided, where structural genes may be introduced in reading frame with the cassette to provide for expression of the natrual peptide, a fused peptide, or a peptide precursor.

Novel methods and compositions are provided for enhanced yield of heterologous proteins in fungi. The method and compositions involve employing fusion sequences involving a sequence encoding a heterologous product produced in relatively large amount as a stable polypeptide in the host fused to a second sequence in open reading frame with the prior sequence coding for a different heterologous polypeptide, where the two polypeptides are joined by a selectively cleavable linkage. In particular, a sequence coding for superoxide dismutase is joined to another polypeptide of interest at either terminus of the superoxide dismutase in a yeast expression vector under transcriptional control of an active promoter and the vector introduced into a yeast host and the host grown. High yields of the fusion product are obtained in this manner, where the fusion product can be selectively cleaved so as to produce both the superoxide dismutase and the other polypeptide in high yield. The S. cerevisiae strain 2150-2-3 (pYASI1) was deposited at the A.T.C.C. on Feb. 27, 1985 and given accession No. 20745. The S. cerevisiae strain AB110 (pYLUIGF2-14) was deposited at the A.T.C.C. on Mar. 19, 1986 and given accession No. 20796.

Growing community of inventors

Walnut Creek, CA, United States of America

Jeffrey R Shuster