Keto-acyl-(ACP) reductase promoter from cuphea lanceolata

Multicellular living organisms and unmodified parts thereof and – Plant – seedling – plant seed – or plant part – per se – Higher plant – seedling – plant seed – or plant part

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435468, 536 241, 800278, A01H 500, C07H 2104, C12N 1582

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active

061335061

DESCRIPTION:

BRIEF SUMMARY
BACKGROUND OF THE INVENTION

On the basis of compartmentalization, the biosynthesis of fatty acids and triacylglycerides may be regarded as separate pathways of biosynthesis, but in view of the end product, as one pathway of biosynthesis. De novo biosynthesis of fatty acids takes place in plastids and is catalyzed by three enzymes or enzyme systems, namely (1) acetyl-CoA carboxylase (ACCase), (2) fatty acid synthase (FAS), and (3) acyl-[ACP] thioesterase (TE). In most organisms the end products of these reaction sequences are either palmitic acid, stearic acid or, after desaturation, oleic acid.
Fatty acid synthase consists of an enzyme complex of dissociable single enzymes with the individual enzymes acetyl-[ACP] transacylase, malonyl-[ACP] transacylase, .beta.-ketoacyl-[ACP] synthases I, II, III, .beta.-ketoacyl-[ACP] reductase, liydroxyacyl-[ACP] dehydratase, enoyl-[ACP] reductase and ACP=acyl carrier protein.
Then, in the so-called Kennedy pathway, triacylglyceride biosynthesis takes place from glycerol 3-phosphate and fatty acids--which are present as acyl-CoA substrates--in the cytoplasm at the endoplasmic reticulum.
The expression of genes of fatty acid biosynthesis is decisively regulated by their upstream promoters. They control the strength of the tissue-specific, development-specific or external stimulus-induced expression of the genes downstream thereto.
A large number of plant promoters, including seed-specific promoters, have been isolated and characterized during the last few years. A few examples are the HMW promoter (L. S. Robert et al., Plant Cell 1, pp. 569-578 (1989); V. Colot et al., Mol. Gen. Genet. 216, pp. 81-90 (1989)), Baumlein et al., The Plant Journal 2, pp. 233-239, 1992; zein promoter (A. J. M. Matzke et al., Plant Mol. Biol. 14, pp. 323-332 (1990)), lectin promoter (P. Guerche et al., Mol. Gen. Genet. pp. 306-314 (1990), USP promoter (H. Baumlein et al., Mol. Gen. Genet. 225, pp. 459-467 (1991)), napin promoter (M. Stayton et al., Aust. J. Plant Physiol. 18, pp. 507-517 (1991), oleosin promoter (J. S. Keddie et al. Plant Mol. Biol. 19, pp. 443-453 (1992)) or ACP promoter (J. de Silva et al., Plant Mol. Biol. 18, pp. 1163-1172 (1992)). The extent to which they are suitable for the expression of a given gene, and the differences which they show with regard to the desired phenotype, cannot be predicted. Frequently the studies of their specificity were carried out on plant species other than the respective cultivated plants of interest. Investigated in rape and found suitable for modifications of the fatty acid metabolism were a napin promoter (J.C. Kridl et al., Seed Sci. Res. 1, pp. 209-219 (1991), D.S. Knutzon et al., Proc. Natl. Acad. Sci. 89, pp. 2624-2628 (1992), and an ACP promoter (Knutzon et al., D. E. Scherer et al., Plant. Mol. Biol. 9, pp. 127-134 (1987)).


SUMMARY OF THE INVENTION

The object of the present invention is first of all to provide promoters with which foreign genes can be expressed in plants with a high efficiency, or be brought to expression in a targeted manner in certain tissues or cell types.
This object is accomplished with the promoters and/or other regulatory elements in the 5' non-translated region according to claim 1.
The invention relates to promoters and/or other regulatory elements in the 5' non-translated region of genes which code for proteins of de novo fatty acid biosynthesis, and alleles and derivatives of these promoters.
Furthermore, the invention relates to genomic clones containing a gene which codes for a protein of de novo fatty acid biosynthesis, and alleles and derivatives of this gene, where the gene comprises the promoter, the structural gene or at least parts thereof, as well as other regulator sequences.
The invention also relates to a process for the preparation of transgenic plants, plant parts and plant products in which a promoter and/or other regulatory elements in the 5' non-translated region of genes coding for proteins of de novo fatty acid biosynthesis are coupled with a desired gene to be expressed, and then trans

REFERENCES:
Baumlein, Helmut, et al., "Cis-analysis of a seed protein gene promoter: the conservative RY repeat CATGCATG within the legumin box is essential for tissue-specific expression of a legumin gene", The Plant Journal, vol. 2, No. 2, pp. 233-239 (1992).
Baumlein, Helmut, eg al., "A novel seed protein gene from Vicia faba is developmentally regulated in transgenic tobacco and Arabidopsis plants", Mol. Gen. Genet, vol. 225, pp. 459-467 (1991).
Colot, Vincent, et al., "Molecular characterization of an active wheat LMW glutenin gene and its relation to other wheat and barley prolamin genes", Mol. Gen. Genet, vol. 216, pp. 81-90 (1989).
de Silva, Jacqueline, et al., "The isolation and functional charaterisation of a B.napus acyl carrier protein 5' flanking region involved in the regulation of seed storage lipid systhesis", Plant Molecular Biology, vol. 18, pp. 1163-1172 (1992).
Keddie, James S., "Cloning and characterisation of an oleosin gene from Brassica napus", Plant Molecular Biology, vol. 19, pp. 443-453 (1992).
Knutzon, Deborah S., "Modification of Brassica seed oil by antisense expression of a stearoyl-acyl carrier protein desaturase gene", Proc. Nat. Acad. Sci., USA, vol. 89, pp. 2624-2628 (Apr. 1992).
Kondo, Hiroki, et al., "Acetyl-CoA carboxylase from Escherichia coli; Gene organization and nucleotide sequence of the biotin carboxylase subunit", Proc. Natl. Acad. Sci. USA, vol. 88, pp. 9730-9733, (Nov. 1991).
Luo, Kiaochun, et al., "Structural features of the acetyl-CoA carboxylase gene: Mechanisms for the generation of mRNAs with 5' end heterogeneity", Proc. Natl. Acad. Sci. USA, vol. 86, pp. 4042-4046 (1989).
Matzke, A.J.M., et al., "Deletion analysis of a zein gene promoter in transgenic tobacco plants", Plant Molecular Biology, vol. 14, pp. 323-332 (1990).
Robert, Laurian S., et al., "Tissue-Specific Expression of a Wheat High Molecular Weight Glutenin Gene in Transgenic Tobacco", The Plant Cell, vol. 1, pp. 569-578 (Jun. 1989).
Scherer, Donna E., et al., "Isolation of a cDNA clone for the acyl carrier protein-I of spinach", Plant Molecular Biology, vol. 9, pp. 127-134 (1987).
Stayton, Mark, et al., "High Level, Seed-specific Expression of Foreign Coding Sequences in Brassica napus", Aust. J. Plant Physiol., vol. 18, pp. 507-517 (1991).
Egli, Margaret A., et al., (Abstract 384), "Cloning and Sequence Analysis of a Maize Acetyl-CoA Caroxylase Gene", Supplement to Plant Physiology (Annual Meeting of the American Society of Plant Physiologists), vol. 102, p. 70 (1993).
Yanai, Yukihiro, et al., (Abstract 382), "RFLP Mapping of an Arabidopsis Acetyl-CoA Carboxylase Gene", Supplement to Plant Physiology (Annual Meeting of the American Society of Plant Physiologists), vol. 102, p. 70 (1993).
Slabas, A. R., et al., (Abstract 119: 221570p) "Biochemistry and molecular biology of lipid biosynthesis in plants: potential for genetic manipulation", Plant Biochem, vol. 119, p. 557 (1993).
Schulte, et al., (Abstract Y 315), "Strategies for the isolation of genes involved in de novo fatty acid biosynthesis", J. Cell. Supp., Crop Improvement via Biotechnology: An International Perspective (Keystone Symposia on Molecular & Cellular Biology), p. 227, Supplement 16F, Apr. 3-16 1992.
Klein et al., Isolation and Characterization of a cDNA from Cuphea lanceolata encoding a Beta-Ketoacyl-ACP Reductase, Mol. Gen. Genet. (1992) 233:122-128.
De Silva J, et al. The isolation and functional characterisation of a B. napus acyl carrier protein 5' flanking region involved in the regulation of seed storage lipid synthesis. Plant Mol. Biol. 18:1163-1172, 1992.
O'Neil J, et al. "Cloning and charaterization of genomic B-ketoacyl-ACP synthase genes from castor." J. Cell. Biochem. 16F: 227, 1992.
Benfey PN, et al. "The cauliflower mosaic virus 35S promoter: Combinatorial regulation of transcription in plants." Science 250: 959-966, Nov. 1990.
Kim Y, et al. "A 20 nucleotide upstream element is essential for the nopaline synthase (nos) promoter activity." Plant Mol. Biol. 24: 10

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