Transgenic plants or algae expressing an AGP enzyme coupled to a

Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters carbohydrate production in the plant

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800263, 800268, 800287, 800296, 800298, 8003172, 435 691, 435 698, 435101, 435194, 435468, 536 236, A01H 500, A01H 1300, C12N 1529, C12N 1582, C12N 1554

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059774371

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BRIEF SUMMARY
FIELD OF THE INVENTION

The present invention relates to a transgenic orgainism.
In particular, the present invention relates to a transgenic starch producing organism having an increased ability to synthesize starch and one that is capable of producing starch in high yields. More particularly the present invention relates to a transgenic organism comprising a nucleotide sequence coding for exogenous ADP glucose pyrophosphorylase (AGP).
In a preferred embodiment the present invention relates to a transgenic plant or plant cell capable of expressing exogenous AGP in the starch producing centers in the plant, namely the chloroplasts and the amyloplasts. The invention also relates to a recombinant DNA construct for use in the transformation of a plants or plant cell showing enhanced starch production, and plants and plant cells transformed with the recombinant DNA construct.
ADP glucose pyrophosphorylase (E.C.2.7.7.27) (AGP) is one of the primary enzymes involved in the biosynthesis of starch and glycogen in organisms such as plants, algae, fungi and bacteria, particularly plants.
AGP catalyses the reaction: biosynthesis of starch in plants. Moreover, that reaction has been shown to be the rate limiting factor in the synthesis of starch in organisms such as plants, the rate of that reaction in turn being critically dependent upon the AGP concentration. Because of that, AGP has been the subject of intensive investigation and for a general review of recent studies on plant AGP, reference should be made to Kleczkowski et al: Z. Naturforsch. 46c, 605-612 (1991).
As reported by Kleczkowski et al (ibid) and elsewhere, AGP is widely distributed throughout the plant kingdom and is found in some starch producing bacteria, such as E. coli. Plant AGP exists as a tetramer (210 to 240 kDa) composed of two small sub-units (50 to 55 kDa) and two large sub-units (51 to 60 kDa) in contrast to bacterial AGP which appears to consist of four units of equal size. AGP has also been shown to be produced in cyanobacteria and in algae, where its tetrameric structure is similar to that in plants, i.e. two large and two small sub-units, rather than the homotetrameric structure found in ordinary bacteria.
Because of the commercial importance of starch, primarily as a foodstuff but also as an important industrial chemical, AGP itself and recombinant DNA constructs containing DNA sequences encoding AGP for the transfection of plants and plant cells as a means of increasing plant AGP concentration and hence increased biosynthesis of starch in plants and increased starch yields, have formed the subject matter of several recently published patent applications.
For example, in EP-A-0368506 a method of extracting AGP from wheat leaf and wheat endosperm is disclosed. Also disclosed are the cDNA sequences encoding wheat leaf and wheat endosperm AGP, and various plasmids containing those sequences for subsequent insertion into plants to provide plants having an increased ability to synthesise starch, although that latter step is not described in detail, nor are any examples given of transgenic plants containing those constructs.
WO 91/19806 discloses transformed plant cells and plants having elevated levels of starch and starch biosynthesis achieved by incorporating into the plant genome a DNA construct comprising in sequence a plant promoter, a DNA sequence encoding a fusion polypeptide consisting of a plastid transit peptide and a bacterial (E. coli) AGP, and a 3'-non-translated region which functions in the plant cell to cause transcriptional termination and the addition of a polyadenylated tail to the 3'- end of the corresponding DNA sequence. The DNA sequence encoding E. coli AGP is given, as well as the deduced amino acid sequence. Transgenic potato and tomato plants transformed with the E. coli AGP gene are shown to produce increased starch yields. It is suggested that other bacterial sources besides E. coli, and also algae, may be used as a source for the AGP gene to be used in the transformation of the plants and plant cells to provide i

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