P450 Monooxygenases of the cyp79 family

A method for the isolation of a cDNA coding for a P450 monooxygenase converting an aliphatic or aromatic amino acid or chain-elongated methionine to the corresponding oxime;

• comprising
• (a) preparing a cDNA library from plant tissue expressing such a monooxygenase,
• (b) using at least one oligonucleotide designed on the basis of SEQ ID NO:
• 1, SEQ ID NO:
• 2, SEQ ID NO:
• 3, or SEQ ID NO:
• 4;
• SEQ ID NO:
• 9, SEQ ID NO:
• 10, SEQ ID NO:
• 11, or SEQ ID NO:
• 12;;
• SEQ ID NO:
• 39 and SEQ ID NO:
• 40;
• SEQ ID NO:
• 54, SEQ ID NO:
• 55, SEQ ID NO:
• 56, SEQ ID NO:
• 70 or SEQ ID NO:
• 71;
• or SEQ ID NO:
• 74, SEQ ID NO:
• 75, SEQ ID NO:
• 84 or SEQ ID NO:
• 85 to amplify part of the P450 monooxygenase cDNA from the cDNA library,
• (c) optionally using a further oligonucleotide designed on the basis of SEQ ID NO:
• 1, SEQ ID NO:
• 2, SEQ ID NO:
• 3, or SEQ ID NO:
• 4;
• SEQ ID NO:
• 9, SEQ ID NO:
• 10, SEQ ID NO:
• 11, or SEQ ID NO:
• 12;;
• SEQ ID NO:
• 39 and SEQ ID NO:
• 40;
• SEQ ID NO:
• 54, SEQ ID NO:
• 55, SEQ ID NO:
• 56, SEQ ID NO:
• 70 or SEQ ID NO:
• 71;
• or SEQ ID NO:
• 74, SEQ ID NO:
• 75, SEQ ID NO:
• 84 or SEQ ID NO:
• 85 to amplify part of the P450 monooxygenase cDNA from the cDNA library in a nested PCR reaction,
• (d) using the DNA obtained in steps (b) or (c) as a probe to screen a cDNA library prepared from plant tissue expressing a P450 monooxygenase converting an aliphatic or aromatic amino acid or chani-elongated methinone honologue to the corresponding oxime, and
• (e) identifying and purifying vector DNA comprising an open reading frame encoding a protein characterized by an amino acid sequence showing at least 40% identity to the amino acid sequence resulting from the global alignment with SEQ ID NO:
• 1 or SEQ ID NO:
• 3 or both;
• SEQ ID NO:
• 39;
• SEQ ID NO:
• 54 or SEQ ID NO:
• 70 or both;
• or at least 50% identity to the amino acid sequence resulting from the global alignment with SEQ ID NO:
• 9 or SEQ ID NO:
• 11 or both;
• or SEQ ID NO:
• 74 or SEQ ID NO:
• 84 or both;
• (f) optionally further processing the purified DNA.