DNA Sequences Encoding Caryophyllaceae and Caryophyllaceae-Like Cyclopeptide Precursors and Methods of Use
OwnerPatrick S. Covello, Raju S.S. Datla +5inventors
InventorsPatrick S. Covello, Raju S.S. Datla, Sandra Lee Stone, J. John Balsevich, Martin John Reaney, Paul Grenville Arnison, Janet Anne Condie
Life of this patent
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Abstract
Naturally-occurring and modified recombinant nucleic acid molecules have been isolated that encode linear pre-cursors of cyclopeptides of the Caryophyllaceae (Ccps) and Caryophyllaceae-like (Clcps) type V1 class of cyclopeptides. Such nucleic acid molecules are useful for producing cyclopeptides and their linear precursors by recombinant methods.
Claims18
A method of producing a cyclopeptide comprising:
- providing a linear polypeptide comprising the amino acid sequence as set forth in SEQ ID NO:
- 3, SEQ ID NO:
- 7, SEQ ID NO:
- 13, SEQ ID NO:
- 16, SEQ ID NO:
- 19, SEQ ID NO:
- 23, SEQ ID NO:
- 26, SEQ ID NO:
- 29, SEQ ID NO:
- 32, SEQ ID NO:
- 37, SEQ ID NO:
- 38, SEQ ID NO:
- 39, SEQ ID NO:
- 41, SEQ ID NO:
- 43, SEQ ID NO:
- 45, SEQ ID NO:
- 47, SEQ ID NO:
- 49 or SEQ ID NO:
- 51;
- and, subjecting the linear polypeptide to conditions under which a cyclopeptide consisting of the amino acid sequence as set forth in SEQ ID NO:
- 3, SEQ ID NO:
- 7, SEQ ID NO:
- 13, SEQ ID NO:
- 16, SEQ ID NO:
- 19, SEQ ID NO:
- 23, SEQ ID NO:
- 26, SEQ ID NO:
- 29, SEQ ID NO:
- 32, SEQ ID NO:
- 37, SEQ ID NO:
- 38, SEQ ID NO:
- 39, SEQ ID NO:
- 41, SEQ ID NO:
- 43, SEQ ID NO:
- 45, SEQ ID NO:
- 47, SEQ ID NO:
- 49 or SEQ ID NO:
- 51 is produced by cyclization of the linear polypeptide.
A method of reducing cyclopeptide content in a host cell, tissue or plant comprising:
- reducing expression in the cell, tissue or plant of a nucleic acid molecule comprising a nucleotide sequence having at least 80% sequence identity to the nucleotide sequence as set forth in SEQ ID NO:
- 1, SEQ ID NO:
- 4, SEQ ID NO:
- 8, SEQ ID NO:
- 10, SEQ ID NO:
- 14, SEQ ID NO:
- 17, SEQ ID NO:
- 20, SEQ ID NO:
- 24, SEQ ID NO:
- 27, SEQ ID NO:
- 30, SEQ ID NO:
- 33, SEQ ID NO:
- 34, a codon degenerate sequence thereof or a full length complement thereof, compared to expression of the nucleotide sequence in the cell, tissue or plant before expression was reduced.
An isolated nucleic acid molecule comprising a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence as set forth in SEQ ID NO:
- 1, SEQ ID NO:
- 4, SEQ ID NO:
- 8, SEQ ID NO:
- 10, SEQ ID NO:
- 14, SEQ ID NO:
- 17, SEQ ID NO:
- 20, SEQ ID NO:
- 24, SEQ ID NO:
- 27, SEQ ID NO:
- 30, SEQ ID NO:
- 33, SEQ ID NO:
- 34, a codon degenerate sequence thereof, or a full length complement thereof.
An isolated linear polypeptide comprising the amino acid sequence as set forth in SEQ ID NO:
- 2, SEQ ID NO:
- 5, SEQ ID NO:
- 6, SEQ ID NO:
- 9, SEQ ID NO:
- 11, SEQ ID NO:
- 12, SEQ ID NO:
- 15, SEQ ID NO:
- 18, SEQ ID NO:
- 21, SEQ ID NO:
- 22, SEQ ID NO:
- 25, SEQ ID NO:
- 28, SEQ ID NO:
- 31, SEQ ID NO:
- 35 or SEQ ID No:
- 36, or an amino acid sequence thereof having a conservative substitution.
An isolated cyclopeptide consisting of the amino acid sequence as set forth in SEQ ID NO:
- 26, SEQ ID NO:
- 29, SEQ ID NO:
- 32, SEQ ID NO:
- 47, SEQ ID NO:
- 49 or SEQ ID NO:
- 51.
(canceled)
A method of identifying a gene or polypeptide related to cyclopeptide production comprising:
- selecting a nucleic acid molecule that is known to encode a reference cyclopeptide;
- identifying a flanking sequence in the nucleic acid molecule or in a linear polypeptide encoded by the nucleic acid molecule, the flanking sequence flanking a nucleotide sequence of the nucleic acid molecule that encodes the reference cyclopeptide or flanking an amino acid sequence of the linear polypeptide that corresponds to the reference cyclopeptide;
- searching a database of nucleic acid molecules or polypeptides for target sequences that have at least 80% sequence identity to the flanking sequence to thereby identify nucleotide or amino acid sequences that correspond to the gene or polypeptide related to cyclopeptide production.
Dependencies are inferred from the claim text.