Nucleic and amino acid sequences for a novel transketolase...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/29 (2006.01) C12N 1/21 (2006.01) C12N 5/14 (2006.01) C12N 9/10 (2006.01) C12N 15/52 (2006.01) C12N 15/54 (2006.01) C12N 15/82 (2006.01)

Patent

CA 2302873

cDNAs encoding 1-deoxyxylulose-5-phosphate synthase from peppermint Mentha piperita have been isolated and sequenced, and the corresponding amino acid sequences have been determined. Accordingly, isolated DNA sequences (SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7) are provided which code for the expression of 1-deoxyxylulose-5- phosphate synthase from essential oil plant species. In other aspect the present invention provides for isolated, recombinant plant DXPS proteins, such as the proteins having the sequences set forth in SEQ ID NO:4, SEQ ID NO:6 and SEQ ID NO:8. In other aspects, replicable recombinant cloning vehicles are provided which code for plant 1-deoxyxylulose-5-phosphate synthases, or for a base sequence sufficiently complementary to at least a portion of 1-deoxyxylulose-5-phosphate synthase DNA or RNA to enable hybridization therewith. In yet other aspects, modified host cells are provided that have been transformed, transfected, infected and/or injected with a recombinant cloning vehicle and/or DNA sequence encoding a plant 1-deoxyxylulose-5-phosphate synthase. Thus, systems and methods are provided for the recombinant expression of the aforementioned recombinant 1-deoxyxylulose-5-phosphate synthase that may be used to facilitate its production, isolation and purification in significant amounts. Recombinant 1-deoxyxylulose-5-phosphate synthase may be used to obtain expression or enhanced expression of 1-deoxyxylulose-5-phosphate synthase in plants in order to enhance the production of 1-deoxyxylulose-5-phosphate, or its derivatives such as isopentenyl diphosphate (IPP), or may be otherwise employed for the regulation or expression of 1-deoxyxylulose-5-phosphate synthase, or the production of its products.

Selon l'invention, on a isolé et séquencé des ADNc codant la 1-désoxyxylulose-5-phosphate synthase provenant de la menthe poivrée (Mentha piperita), et on a déterminé les séquences d'acides aminés correspondantes. En conséquence, l'invention concerne des séquences isolées d'ADN (SEQ ID N0:3, SEQ ID N0:5, SEQ ID N0:7) qui codent pour l'expression de la 1-désoxyxylulose-5-phosphate synthase provenant d'une espèce végétale à huile essentielle. Dans un autre aspect, l'invention concerne des protéines DXPS de plante, isolées et recombinées, telles que les protéines possédant les séquences formulées dans SEQ ID NO:4, SEQ ID NO:6 et SEQ ID NO:8. Dans d'autres aspects, l'invention concerne des supports de clonage de recombinaison et de réplication, qui codent pour les 1-désoxyxylulose-5-phosphate synthases végétales, ou pour une séquence de base suffisamment complémentaire à au moins une portion de l'ADN ou de l'ARN de la 1-désoxyxylulose-5-phosphate synthase, afin de permettre l'hybridation avec celles-ci. Dans encore d'autres aspects, l'invention concerne des cellules hôtes modifiées qui ont été transformées, transfectées, infectées et/ou injectées au moyen d'un support de clonage recombiné et/ou d'une séquence d'ADN codant une 1-désoxyxylulose-5-phosphate synthase végétale. Ainsi, l'invention concerne des systèmes et procédés destinés à l'expression recombinée de la synthase recombinée ci-dessus mentionnée, que l'on peut utiliser pour faciliter la production, isolation et purification de cette synthase, en quantités importantes. On peut utiliser également cette synthase recombinée pour obtenir l'expression ou l'expression accrue de la 1-désoxyxylulose-5-phosphate synthase dans des plantes, de manière à accroître la production de 1-désoxyxylulose-5-phosphate, ou de ses dérivés comme le diphosphate d'isopentényle (IPP), ou on peut encore l'employer pour la régulation ou l'expression de la 1-désoxyxylulose-5-phosphate synthase, ou pour la production des produits de celle-ci.

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