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AssigneeChiron Corporation
InventorsRae Lyn Burke, Steven Rosenberg, Jeffrey R. Shuster, Patricia A. Tekamp-Olson, Pablo D. T. Valenzuela
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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.
A DNA construct comprising in order of transcription a first transcription regulatory region obtained from one of the yeast genes ADR3, the regulatory region of PHO5, or the regions regulated by GAL4, which provides for inducible transcriptional regulation;
A DNA construct comprising in order of transcription a first transcriptional regulatory region obtained from one of the yeast genes ADR3, the regulatory region of PHO5, or the regions regulated by GAL4, which provides for inducible transcriptional regulation;
An expression vector comprising:
A DNA construct comprising in order of transcription a first transcriptional regulatory region obtained from one of the yeast genes ADR3, the regulatory region of PHO5, or the regions regulated by GAL4;
A transcriptional regulation control region comprising in the direction of transcription a transcriptional regulatory region obtained from one of the yeast genes ADR3, the regulatory region of PHO5, or the regions regulated by GAL4, and a yeast transcription initiation region from the yeast glycolytic enzyme GAPDH of FIG. 2, as a linear fragment or in combination with a DNA sequence functional in a yeast host.
Dependencies are inferred from the claim text.