Multicellular living organisms and unmodified parts thereof and – Method of introducing a polynucleotide molecule into or... – The polynucleotide alters carbohydrate production in the plant
Patent
1997-04-17
2000-11-14
Fox, David T.
Multicellular living organisms and unmodified parts thereof and
Method of introducing a polynucleotide molecule into or...
The polynucleotide alters carbohydrate production in the plant
800286, 800287, 8003172, 435101, 4353201, 435468, 536 236, 536 241, 536 245, C12N 1529, C12N 1582, A01H 500, C12P 1904
Patent
active
061472794
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to a method of inhibiting gene expression, particularly inhibiting gene expression in a plant. The present invention also relates to a nucleotide sequence useful in the method. In addition, the present invention relates to a promoter that is useful for expressing the nucleotide sequence.
Starch is one of the main storage carbohydrates in plants, especially higher plants. The structure of starch consists of amylose and amylopectin. Amylose consists essentially of straight chains of .alpha.-1-4-linked glycosyl residues. Amylopectin comprises chains of .alpha.-1-4-linked glycosyl residues with some .alpha.-1-6 branches. The branched nature of amylopectin is accomplished by the action of inter alia an enzyme commonly known as the starch branching enzyme ("SBE"). SBE catalyses the formation of branch points in the amylopectin molecule by adding .alpha.-1,4 glucans through .alpha.-1,6-glucosidic branching linkages. The biosynthesis of amylose and amylopectin is schematically shown in FIG. 1, whereas the .alpha.-1-4-links and the .alpha.-1-6 links are shown in FIG. 2.
It is known that starch is an important raw material. Starch is widely used in the food, paper, and chemical industries. However, a large fraction of the starches used in these industrial applications are post-harvest modified by chemical, physical or enzymatic methods in order to obtain starches with certain required functional properties.
Within the past few years it has become desirable to make genetically modified plants which could be capable of producing modified starches which could be the same as the post-harvest modified starches. It is also known that it may be possible to prepare such genetically modified plants by expression of antisense nucleotide coding sequences. In this regard, June Bourque provides a detailed summary of antisense strategies for the genetic manipulations in plants (Bourque 1995 Plant Science 105 pp 125-149). At this stage, reference could be made to FIG. 3 which is a schematic diagram of one of the proposed mechanisms of antisense-RNA inhibition.
In particular, WO 92/11375 reports on a method of genetically modifying potato so as to form amylose-type starch. The method involves the use of an anti-sense construct that can apparently inhibit, to a varying extent, the expression of the gene coding for formation of the branching enzyme in potato. The antisense construct of WO 92/11375 consists of a tuber specific promoter, a transcription start sequence and the first exon of the branching enzyme in antisense direction. However, WO 92/11375 does not provide any antisense sequence data. In addition, WO 92/11375 only discloses the use of the potato GBSS promoter.
WO 92/14827 reports on a plasmid that, after insertion into the genome of a plant, can apparently cause changes in the carbohydrate concentration and carbohydrate composition, such as the concentration and composition of amylose and amylopectin, in the regenerated plant. The plasmid contains part of the coding sequence of a branching enzyme in an antisense orientation.
EP-A-0647715 reports on the use of antisense endogenous mRNA coding DNA to alter the characteristics and the metabolic pathways of ornamental plants.
EP-A-0467349 reports on the expression of sequences that are antisense to sequences upstream of a promoter to control gene expression.
EP-A-0458367 and U.S. Pat. No. 5,107,065 report on the expression of a nucleotide sequence to regulate gene expression in a plant. The nucleotide sequence is complementary to a mRNA sequence of a gene and may cover all or a portion of the non-coding region of the gene. In other words, the nucleotide sequences of EP-A-0458367 and U.S. Pat. No. 5,107,065 must at least comprise a sequence that is complementary to a coding region. EP-A-0458367 and U.S. Pat. No. 5107065 contain minimal sequence information.
Kuipers et al in Mol. Gen. Genet. [1995] 246 745-755 report on the expression of a series of nucleotides that are antisense to part of the genomic intron sequences of p
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Danisco A/S
Fox David T.
Kowalski Thomas J.
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