Genes and proteins for aromatic polyketide synthesis
AssigneesNational Research Council of Canada, University of Saskatchewan
InventorsJonathan E. Page, Steve Gagne
Life of this patent
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Abstract
Nucleic acid molecules encoding polypeptides having polyketide synthase activity have been identified and characterized. Expression or over-expression of the nucleic acids alters levels of cannabinoid compounds in organisms. The polypeptides may be used in vivo or in vitro to produce cannabinoid compounds.
Claims25
An isolated or purified cDNA nucleic acid molecule consisting of:
- i) a nucleotide sequence as set forth in SEQ ID NO:
- 1 or 3 or a codon degenerate nucleotide sequence thereof;
- ii) a nucleotide sequence of 303 nucleotides that encodes a polypeptide with an amino acid sequence of SEQ ID NO:2 or a conservatively substituted amino acid sequence thereof;
- or iii) a fragment of i) or ii) having a length of at least 300 base pairs;
- wherein the nucleic acid molecule encodes a polypeptide with polyketide cyclase activity.
An isolated or purified polypeptide consisting of:
- i) an amino acid sequence having one amino acid difference from the sequence set forth in SEQ ID NO:2;
- ii) a conservatively substituted amino acid sequence of the sequence set out in SEQ ID NO:2;
- iii) any of i) or ii) tagged with a HIS tag;
- or iv) an amino acid sequence as set forth in SEQ ID NO:2 tagged with a HIS tag.
A process of altering levels of cannabinoid compounds in a cannabis plant, cannabis cell or cannabis tissue comprising introducing a nucleic acid molecule using a RNAi construct, a miRNA, VIGS virus, antisense oligonucleotide, a targeted mutagenesis construct or using Targeting Induced Local Lesions IN Genomes (TILLING) wherein the nucleic acid molecule or TILLING inhibits expression of SEQ ID NO:1 or a part thereof encoding a polypeptide with polyketide cyclase activity that catalyzes synthesis of an aromatic polyketide, wherein the altered levels of cannabinoid compounds is in comparison to cannabis plant, cannabis cell or cannabis tissue of the same species grown under similar conditions but that does not comprise the nucleic acid molecule for inhibiting SEQ ID NO:
- 1 or part thereof, or that has not been subjected to TILLING.
A process of altering levels of cannabinoid compounds in an organism, cell or tissue comprising introducing and expressing or over-expressing a nucleic acid molecule encoding a polypeptide having at least 95% sequence identity to SEQ ID NO:
- 2 or a conservatively substituted amino acid sequence of the sequence set forth in SEQ ID NO:
- 2, and having polyketide cyclase activity in the organism, cell or tissue, wherein the expressing or over-expression is in comparison to an organism, cell or tissue of the same species grown under similar conditions but without the introducing and expressing or over-expressing of the nucleic acid molecule.
Dependencies are inferred from the claim text.