Method of producing recombinant sucrose synthase, use...

Details Method of producing recombinant sucrose synthase, use... Method of producing recombinant sucrose synthase, use...

C - Chemistry, Metallurgy

12

N

C12N 15/29 (2006.01) A01H 5/00 (2006.01) C12N 15/70 (2006.01) C12N 15/82 (2006.01) C12Q 1/48 (2006.01)

Patent

CA 2554780

The invention relates to a method for the efficient production of large quantities of soluble recombinant sucrose synthase (SS) in the active form thereof, with the expression of the gene that codes for SS in a strain of Escherichia coli. The expression vector used enables the recombinant SS thus produced to have a histidine tail which facilitates the rapid purification thereof. In addition, the invention relates to sequences of mutated versions of the SS gene, which code for SS isoforms that are suitable for the production of ADPG. The invention also relates to an efficient method for the production of ADPG and UDPG, using the wild and mutated versions of recombinant SS. The invention further relates to the use of SS for the production of sucrose determination test devices. Finally, the invention relates to a method of obtaining transgenic plants which overexpress the SS gene, either constituently or in the reserve organs or leaves thereof, and which have a high concentration (both in the leaves and in the reserve tissues) of sucrose, ADPG, G6P and starch owing to the high ADPG-synthesising activity of SS.

Un procédé de production efficace de grandes quantités de SS recombinante soluble sous sa forme active au moyen de l'expression du gène codant pour la SS dans une souche de Escherichia coli. Le vecteur d'expression utilisé permet que la SS recombinante obtenue possède une queue d'histidines facilitant sa purification de manière rapide. Sont également décrites les séquences de version mutées du gène de la SS codant pour des isoformes de la SS adéquates pour la production de ADPG. Grâce à l'utilisation des versions "sylvestre" et "mutée" de SS recombinante, on décrit un procédé efficace de production d'ADPG et UDPG. On décrit également l'utilisation de la SS pour la production de dispositifs d'essai de détermination de saccharose. Finalement, on décrit la description de plantes transgéniques qui surexpriment le gène de la SS, soit par constitution, soit sous forme de feuilles ou organes de réserve, et qui possèdent une forte teneur (en feuilles et en tissus de réserve) en saccharose, ADPG, G6P et amidon comme résultat de la forte activité de synthèse d'ADPG de la SS.

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