Cytochrome P450S and uses thereof
AssigneeUniversity of Kentucky Research Foundation
InventorsJoseph Chappell, Lyle F. Ralston
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Abstract
The invention relates to isolated cytochrome P450 polypeptides and nucleic acid molecules, as well as expression vectors and transgenic plants containing these molecules. In addition, the invention relates to uses of such molecules in methods of increasing the level of resistance against a disease caused by a plant pathogen in a transgenic plant, in methods for producing altered compounds, for example, hydroxylated compounds, and in methods of producing isoprenoid compounds.
Claims15
A method for producing in a recombinant host cell an isoprenoid compound not endogenously produced by a non recombinant host cell, comprising:
- a) providing a recombinant host cell that comprises heterologous nucleic acid encoding an isoprenoid synthase, and heterologous nucleic acid encoding one or more protein(s) comprising a CYP71, CYP73, CYP82 or a CYP92 family cytochrome P450 polypeptide encoded by nucleic acid that can be amplified with degenerate primers based on any one of SEQ ID NOs:
- 26-29, wherein:
- the isoprenoid synthase catalyzes production of an isoprenoid compound;
- wherein the isoprenoid synthase is a sesquiterpene synthase;
- and
- the cytochrome P450 polypeptide(s) catalyzes dual hydroxylation, oxidation, demethylation or methylation of the isoprenoid compound;
- and
- b) culturing the recombinant host cell under conditions suitable for expressing the isoprenoid synthase and the cytochrome P450 polypeptide(s) under conditions for producing the isoprenoid compound wherein the synthase and the cytochrome P450 polypeptide(s) catalyze formation of the isoprenoid compound in the host cell.
A host cell, comprising nucleic acid encoding an isoprenoid synthase and nucleic acid encoding one or more protein(s) comprising a CYP71, CYP73, CYP82 or a CYP92 family cytochrome P450 polypeptides encoded by nucleic acid that can be amplified with degenerate primers based on any one of SEQ ID NOs:
- 26-29, wherein:
- the nucleic acid encoding the synthase and the nucleic acid encoding the P450 polypeptides are heterologous to the host cell;
- the synthase catalyzes production of an isoprenoid compound;
- the isoprenoid synthase is a sesquiterpene synthase;
- the P450 polypeptides catalyze hydroxylation, oxidation, demethylation or methylation of the isoprenoid compound;
- at least one of the P450 polypeptide(s) catalyzes the dual hydroxylation, oxidation, demethylation or methylation of the isoprenoid compound whose production is catalyzed by the synthase;
- and
- the isoprenoid compound produced by the host cell is not endogenously produced by a non recombinant host cell.
Dependencies are inferred from the claim text.