Dna sequences for enzymatic synthesis of polyketide or...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/52 (2006.01) C07K 14/195 (2006.01) C12N 5/10 (2006.01) C12N 9/00 (2006.01) C12N 15/63 (2006.01) C12P 7/62 (2006.01) C12P 17/06 (2006.01) C12P 17/18 (2006.01)

Patent

CA 2346499

The invention consists of: (1) cloned Sorangium cellulosum polyketide synthase (PKS) biosynthetic cluster DNA; and (2) the nucleotide sequence and predicted protein coding sequences of the cloned DNA. The invention can be used for, but not limited to: (a) increasing yields of PKS product in Sorangium cellulosum (e.g., by amplification or genetic modification of the epothilone gene cluster or its component parts); (b) increasing yields of polyketide product in a heterologous system by transfer of the epothilone gene cluster or its component parts, which may be followed by amplification or genetic modification of the PKS gene cluster or its component parts; (c) modification of the polyketide product chemical structure in either Sorangium cellulosum or a heterologous host (e.g., by genetic modification of the epothilone gene cluster or its component parts; and (d) for the detection of genes and gene products involved in making polyketides or related molecules in other organisms (e.g., by hybridization or complementation assays). DNA sequence and analysis is presented for the following cosmids and plasmids: A2 cosmid; the pEPOcos6 region (overlapping of pEPOcos6 and pEPOcos7); pEPOcos8 cosmid; A5 cosmid; Sau4 (10 kb plasmid).

L'invention concerne: (1) un ADN biosynthétique cloné en grappe de polykétide synthase (PKS) de Sorangium cellulosum; et (2) la séquence nucléotidique et les séquences protéiques codantes prévues de l'ADN cloné. L'invention peut avoir les applications suivantes (sans caractère limitatif): (a) augmentation de la production de PKS chez Sorangium cellulosum (p.ex., par l'amplification ou la modification génétique de la grappe de gènes épothilone ou de ces parties constitutives); (b) augmentation de la production du produit polykétide dans un système hétérologue par le transfert de la grappe de gènes épothilone ou de ces parties constitutives, qui peut être suivie par l'amplification ou la modification génétique de la grappe de gènes PKS ou des ses parties constitutives; (c) modification de la structure chimique du produit polykétide soit chez Sorangium cellulosum soit chez un hôte hétérologue (p.ex., par l'amplification ou la modification génétique de la grappe de gènes épothilone ou de ces parties constitutives); et (d) détection de gènes et de produits géniques participant à la fabrication de polykétides ou de molécules correspondantes dans d'autres organismes (p.ex., par des dosages à hybridation ou à complémentation). La séquence d'ADN et l'analyse sont présentées pour les cosmides et les plasmides suivants: cosmide A2; région pEPOcos6 (se chevauchant avec pEPOcos6 et pEPOcos7); cosmide pEPOcos8; cosmide A5; Sau4 (plasmide 10 kb).

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