Method of producing recombinant sucrose synthase, use thereof in the production of sucrose-determination kits, method of producing ADPglucose and method of obtaining transgenic plants having leaves and reserve organs which accumulate a high concentration of ADPglucose and starch

摘要

Method of production of recombinant sucrose synthase, use thereof in the manufacture of kits for determination of sucrose, production of ADPglucose and production of transgenic plants whose leaves and storage organs accumulate high contents of ADPglucose and starch A method is described for efficient production of large quantities of soluble recombinant SS in its active form, by expression of the gene that encodes SS in a strain of Escherichia coli. The expression vector used means that the recombinant SS produced possesses a histidine tail which facilitates its quick purification. In addition it describes sequences of mutated versions of the gene of SS that encode isoforms of SS suitable for the production of ADPG. Making use of the "wild-type" and "mutated" versions of recombinant SS, an efficient method is described for production of ADPG and UDPG. It also describes the use of SS for the production of assay kits for the determination of sucrose. Finally, it describes the production of transgenic plants which overexpress the gene of SS, either constitutively, or in leaves or storage organs, and which have a high content (both in leaves and in storage tissues) of sucrose, ADPG, G6P and starch as a result of the high ADPG-synthesizing activity of SS.