Expression of higher eucaryotic genes in aspergillus
AssigneeZymoGenetics, Inc.
InventorsGary L. McKnight, Alan Upshall
Life of this patent
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Abstract
A method for expressing higher eucaryotic genes in Aspergillus through the use of a recombinant plasmid capable of integration into the chromosomal DNA of Aspergillus is disclosed. It is preferred to utilize a transcriptional promoter within a DNA construct contained in the plasmid that is of a DNA sequence encoding an ADH enzyme or a TPI enzyme. Promoters capable of directing the expression of a heterologous gene in Aspergillus, as well as other filamentous fungal genera are also disclosed.
Claims10
A DNA construct capable of directing the expression of higher eucaryotic genes in Aspergillus, said DNA construct containing a transcriptional promoter selected from the group consisting of the Aspergillus nidulans ADH3 promoter, the Aspergillus nidulans tpiA promoter, the Aspergillus niger adhA promoter and the Aspergillus niger tpiA promoter operably linked to a higher eucaryotic gene, said gene operably linked to a terminator selected from the group consisting of the Aspergillus nidulans ADH3 terminator, the Aspergillus nidulans tpiA terminator, the Aspergillus niger adhA terminator and the Aspergillus niger tpiA terminator.
A recombinant plasmid capable of integration into the chromosomal DNA of Aspergillus, said plasmid containing a DNA construct capable of directing the expression of higher eucaryotic genes in Aspergillus, said DNA construct containing a transcriptional promoter selected from the group consisting of the Aspergillus nidulans ADH3 promoter, the Aspergillus nidulans tpiA promoter, the Aspergillus niger adhA promoter and the Aspergillus niger tpiA promoter operably linked to a higher eucaryotic gene, said gene operably linked to a terminator selected from the group consisting of the Aspergillus nidulans ADH3 terminator, the Aspergillus nidulans tpiA terminator, the Aspergillus niger adhA terminator and the Aspergillus niger tpiA terminator.
An Aspergillus culture transformed with a DNA construct capable of directing the expression of higher eucaryotic genes in Aspergillus, said DNA construct containing a transcriptional promoter selected from the group consisting of the Aspergillus nidulans ADH.sub.3 promoter, the Aspergillus nidulans tpiA promoter, the Aspergillus niger adhA promoter and the Aspergillus niger tpiA promoter operably linked to a higher eucaryotic gene, said gene operably linked to a terminator selected from the group consisting of the Aspergillus nidulans ADH3 terminator, the Aspergillus nidulans tpiA terminator, the Aspergillus niger adhA terminator and the Aspergillus niger tpiA terminator, said culture producing the protein product of said higher eucaryotic gene in recoverable amounts.
A method of producing protein via expression of higher eucaryotic genes in Aspergillus, comprising:
- introducing into an Aspergillus host a recombinant plasmid capable of integration into the chromosomal DNA of Aspergillus, said plasmid containing a DNA construct capable of directing the expression of higher eucaryotic genes in Aspergillus, said DNA construct containing a transcriptional promoter selected from the group consisting of the Aspergillus nidulans ADH3 promoter, the Aspergillus nidulans tpiA promoter, the Aspergillus niger adhA promoter and the Aspergillus niger tpiA promoter operably linked to a higher eucaryotic gene, said gene operably linked to a terminator selected from the group consisting of the Aspergillus nidulans ADH3 terminator, the Aspergillus nidulans tpiA terminator, the Aspergillus niger adhA terminator and the Aspergillus niger tpiA terminator;
- growing said Aspergillus host in an appropriate medium so that said host produces the protein product of said higher eucaryotic gene in recoverable amounts;
- and
- isolating the protein product of said higher eucaryotic gene from said Aspergillus host.
A transcriptional promoter capable of directing the expression of a heterologous gene in Aspergillus wherein said promoter is selected from the group consisting of the Aspergillus nidulans ADH3 promoter, the Aspergillus nidulans tpiA promoter, the Aspergillus niger adhA promoter and the Aspergillus niger tpiA promoter.
Dependencies are inferred from the claim text.