C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/12 (2006.01) C07H 21/02 (2006.01) C07H 21/04 (2006.01) C12N 1/20 (2006.01) C12N 5/00 (2006.01) C12N 9/00 (2006.01) C12N 15/00 (2006.01) C12N 15/52 (2006.01)
Patent
CA 2380639
A cluster of genes involved in the synthesis and/or addition of oleandrose to avermectin aglycones has been cloned. A 11-kb <i>Pst</i>I clone complemented 28 avermectin glycosylation mutants in seven complementation classes. Sequencing of an 10-kb region identified 9 ORFs and an additional partial ORF. Eight of the ORFs were correlated to the seven glycosylation complementation classes. Sequence comparison to Genbank databases identified 6 genes: dTDP- glucose synthase; dTDP-glucose 4,6 dehydrase; dTDP-4-keto-hexose reductase; dTDP-hexose 3,5 epimerase; dTDP-hexose 3' O-methylase; and an avermectin aglycone-dTDP-oleandrose glycosyltransferase. The ninth ORF was essential for biosynthesis of the avermectin aglycones. The partial ORF encoded part of an avermectin polyketide synthase module 7.
L'invention concerne le clonage d'un groupe de gènes impliqués dans la synthèse ou l'addition d'oléandrose à des aglycones d'avermectine. Selon l'invention, un clone <i>Pst</i>I de 11 Kb a complémenté 28 mutants par glycosylation d'avermectine dans sept classes de complémentation. Le séquençage d'une région de 10 Kb a identifié 9 ORF et un ORF partiel supplémentaire. Huit de ces ORF étaient corrélés aux sept classes de complémentation par glycosylation. La comparaison des séquences avec les bases de données Genbank a identifié 6 gènes : dTDP-glucose-synthase, dTDP-glucose-4,6-déhydratase, dTDP-4-céto-hexose-réductase, dTDP-hexose-3,5-épimérase, dTDP-hexose-3'-O-méthylase et une avermectine-aglycone-dTDP-oléandrose-glycosyltransférase. Le neuvième ORF était essentiel pour la biosynthèse des aglycones d'avermectine. L'ORF partiel a encodé une partie d'un module 7 d'avermectine-polycétide-synthase.
Gewain Keith M.
Macneil Douglas J.
Occi James
Merck & Co. Inc.
Ogilvy Renault Llp/s.e.n.c.r.l.,s.r.l.
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