AU2011253658A1 - Arabidopsis genes encoding proteins involved in sugar and lipid metabolism and methods of use - Google Patents

Abstract

Isolated nucleic acids and polypeptides associated with lipid and sugar metabolism regulation are provided. In particular, lipid metabolism proteins (LMP) and encoding nucleic acids originating from Arabidopsis thaliana are provided. The nucleic acids and polypeptides are used in methods of producing transgenic plants and modulating levels of seed storage compounds in a plant. Preferably, the seed storage compounds are lipids, fatty acids, starches, or seed storage proteins.

Description

AUSTRALIA Patents Act 1990 MICHIGAN STATE UNIVERSITY COMPLETE SPECIFICATION STANDARD PATENT Invention Title: Arabidopsis genes encoding proteins involved in sugar and lipid metabolism and methods of use The following statement is a full description of this invention including the best method of performing it known to us: - 1 - ARABIDOPSIS GENES ENCODING PROTEINS INVOLVED IN SUGAR AND LIPID METABOLISM AND METHODS OF USE This is a divisional of AU 2005286889, the entire contents of which are 5 incorporated herein by reference. BACKGROUND OF THE INVENTION Field of the Invention 100011 This invention relates generally to nucleic acid sequences encoding 10 proteins that are related to the presence of seed storage compounds in plants. More specifically, the present invention relates to nucleic acid sequences encoding sugar and lipid metabolism regulator proteins and the use of these sequences in transgenic plants. The invention further relates to methods of applying these novel plant polypeptides to the identification and stimulation of plant growth and/or to the increase of yield of seed 15 storage compounds. Background Art [00021 The study and genetic manipulation of plants has a long history that began even before the famed studies of Gregor Mendel. In perfecting this science, scientists 20 have accomplished modification of particular traits in plants ranging from potato tubers having increased starch content to oilseed plants such as canola and sunflower having increased or altered fatty acid content. With the increased consumption and use of plant oils, the modification of seed oil content and seed oil levels has become increasingly widespread (e.g. T6pfer et al., 1995, Science 268:681-686). Manipulation 25 of biosynthetic pathways in transgenic plants provides a number of opportunities for molecular biologists and plant biochemists to affect plant metabolism giving rise to the production of specific higher-value products. The seed oil production or composition has been altered in numerous traditional oilseed plants such as soybean (U.S. Patent No. 5,955,650), canola (U.S. Patent No. 5,955,650), sunflower (U.S. Patent No. 30 6,084,164), and rapeseed (T6pfer et al., 1995, Science 268:681-686), and non traditional oilseed plants such as tobacco (Cahoon et al., 1992, Proc. Nati. Acad. Sci. USA 89:11184-11188). 100031 Plant seed oils comprise both neutral and polar lipids (See Table 1). The neutral lipids contain primarily triacylglycerol, which is the main storage lipid that 35 accumulates in oil bodies in seeds. The polar lipids are mainly found in the various membranes of the seed cells, e.g. the endoplasmic reticulum, microsomal membranes and the cell membrane. The neutral and polar lipids contain several common fatty acids (See Table 2) and a range of less common fatty acids. The fatty acid composition of - 1A membrane lipids is highly regulated and only a select number of fatty acids are found in membrane lipids. On the other hand, a large number of unusual fatty acids can be incorporated into the neutral storage lipids in seeds of many plant species (Van de Loo et al., 1993, Unusual Fatty Acids in Lipid Metabolism in Plants pp. 91-126, editor TS 5 Moore Jr. CRC Press; Millar et al., 2000, Trends Plant Sci. 5:95-101). Lipids indicated by an asterisk in Table 2 do not normally occur in plant seed oils, but their production in transgenic plant seed oil is of importance in plant biotechnology. Table 1 Plant Lipid Classes Neutral Lipids Triacylglycerol (TAG) Diacylglycerol (DAG) Monoacyl glycerol (MAG) Polar Lipids Monogalactosyldiacylglycerol (MGDG) Digalactosyldiacylglycerol (DGDG) Phosphatidylglycerol (PG) Phosphatidylcholine (PC) Phosphatidylethanolamine (PE) Phosphatidylinositol (PI) Phosphatidylserine (PS) Sulfoguinovosyldiacylglycerol 10 Table 2 Common Plant Fatty Acids 16:0 Palmitic acid 16:1 Palmitoleic acid 16:3 Palmitolenic acid 18:0 Stearic acid 18:1 Oleic acid 18:2 Linoleic acid 18:3 Linolenic acid r18:3 Gamma-linolenic acid* 0:0 Arachidic acid 0:1 Eicosenoic acid 2:6 Docosahexanoic acid (DHA) * 0:2 Eicosadienoic acid 0:4 Arachidonic acid (AA) * 0:5 icosapentaenoic acid (EPA) * 2:1 Erucic acid [0004] Lipids are synthesized from fatty acids and their synthesis may be divided into 15 two parts: the prokaryotic pathway and the eukaryotic pathway (Browse et al., 1986, Biochemical J. 235:25-31; Ohlrogge & Browse, 1995, Plant Cell 7:957-970). The -2prokaryotic pathway is located in plastids that are the primary site of fatty acid biosynthesis. Fatty acid synthesis begins with the conversion of acetyl-CoA to malonyl CoA by acetyl-CoA carboxylase (ACCase). Malonyl-CoA is converted to malonyl-acyl carrier protein (ACP) by the malonyl-CoA:ACP transacylase. The enzyme beta-keto 5 acyl-ACP-synthase III (KAS III) catalyzes a condensation reaction in which the acyl group from acetyl-CoA is transferred to malonyl-ACP to form 3-ketobutyryl-ACP. In a subsequent series of condensation, reduction and dehydration reactions the nascent fatty acid chain on the ACP cofactor is elongated by the step-by-step addition (condensation) of two carbon atoms donated by malonyl-ACP until a 16- or 18-carbon saturated fatty 10 acid chain is formed. The plastidial delta-9 acyl-ACP desaturase introduces the first unsaturated double bond into the fatty acid. Thioesterases cleave the fatty acids from the ACP cofactor and free fatty acids are exported to the cytoplasm where they participate as fatty acyl-CoA esters in the eukaryotic pathway. In this pathway the fatty acids are esterified by glycerol-3-phosphate acyltransferase and lysophosphatidic acid 15 acyltransferase to the sn-l and sn-2 positions of glycerol-3-phosphate, respectively, to yield phosphatidic acid (PA). The PA is the precursor for other polar and neutral lipids, the latter being formed in the Kennedy pathway (Voelker, 1996, Genetic Engineering ed.:Setlow 18:111-113; Shanklin & Cahoon, 1998, Annu. Rev. Plant Physiol. Plant Mol. Biol. 49:611-641; Frentzen, 1998, Lipids 100:161-166; Millar et al., 2000, Trends Plant 20 Sci. 5:95-101). 100051 Storage lipids in seeds are synthesized from carbohydrate-derived precursors. Plants have a complete glycolytic pathway in the cytosol (Plaxton, 1996, Annu. Rev. Plant Physiol. Plant Mol. Biol. 47:185-214) and it has been shown that a complete pathway also exists in the plastids of rapeseeds (Kang & Rawsthorne, 1994, Plant J. 25 6:795-805). Sucrose is the primary source of carbon and energy, transported from the leaves into the developing seeds. During the storage phase of seeds, sucrose is converted in the cytosol to provide the metabolic precursors glucose-6-phosphate and pyruvate. These are transported into the plastids and converted into acetyl-CoA that serves as the primary precursor for the synthesis of fatty acids. Acetyl-CoA in the plastids is the 30 central precursor for lipid biosynthesis. Acetyl-CoA can be formed in the plastids by different reactions and the exact contribution of each reaction is still being debated (Ohlrogge & Browse, 1995, Plant Cell 7:957-970). It is accepted, however, that a large part of the acetyl-CoA is derived from glucose-6-phospate and pyruvate that are imported from the cytoplasm into the plastids. Sucrose is produced in the source organs (leaves, or -3anywhere that photosynthesis occurs) and is transported to the developing seeds that are also termed sink organs. In the developing seeds, sucrose is the precursor for all the storage compounds, i.e. starch, lipids and partly the seed storage proteins. Therefore, it is clear that carbohydrate metabolism in which sucrose plays a central role is very important 5 to the accumulation of seed storage compounds. [0006] Storage compounds such as triacylglycerols (seed oil) serve as carbon and energy reserves, which are used during germination and growth of the young seedling. Seed (vegetable) oil is also an essential component of the human diet and a valuable commodity providing feed stocks for the chemical industry. A mutant of Arabidopsis 10 affected in seed storage compound metabolism is wrinkled) (wril) (Focks & Benning, 1998, Plant Physiol. 118:91-101). The mutant is characterized by an 80% reduction in seed oil content. [0007] Although the lipid and fatty acid content of seed oil can be modified by the traditional methods of plant breeding, the advent of recombinant DNA technology has 15 allowed for easier manipulation of the seed oil content of a plant, and in some cases, has allowed for the alteration of seed oils in ways that could not be accomplished by breeding alone (See, e.g., T6pfer et al, 1995, Science 268:681-686). For example, introduction of a A 12 -hydroxylase nucleic acid sequence into transgenic tobacco resulted in the introduction of a novel fatty acid, ricinoleic acid, into the tobacco seed oil (Van de Loo et 20 al., 1995, Proc. Natl. Acad. Sci USA 92:6743-6747). Tobacco plants have also been engineered to produce low levels of petroselinic acid by the introduction and expression of an acyl-ACP desaturase from coriander (Cahoon et al., 1992, Proc. Natl. Acad. Sci USA 89:11184-11188). [0008] The modification of seed oil content in plants has significant medical, 25 nutritional, and economic ramifications. With regard to the medical ramifications, the long chain fatty acids (C18 and longer) found in many seed oils have been linked to reductions in hypercholesterolemia and other clinical disorders related to coronary heart disease (Brenner, 1976, Adv. Exp. Med. Biol. 83:85-101). Therefore, consumption of a plant having increased levels of these types of fatty acids may reduce the risk of heart 30 disease. Enhanced levels of seed oil content also increase large-scale production of seed oils and thereby reduce the cost of these oils. [00091 In order to increase or alter the levels of compounds such as seed oils in plants, nucleic acid sequences and proteins regulating lipid and fatty acid metabolism -4- 6 must be identified. As mentioned earlier, several desaturase nucleic acids such as the A desaturase nucleic acid, A 12 -desaturase nucleic acid, and acyl-ACP desaturase nucleic acid have been cloned and demonstrated to encode enzymes required for fatty acid synthesis in various plant species. Oleosin nucleic acid sequences from such different 5 species as Brassica, soybean, carrot, pine and Arabidopsis thaliana also have been cloned and determined to encode proteins associated with the phospholipid monolayer membrane of oil bodies in those plants. 100101 It has also been determined that two phytohornones, gibberellic acid (GA) and absisic acid (ABA), are involved in overall regulatory processes in seed development 10 (e.g. Ritchie & Gilroy, 1998, Plant Physiol. 116:765-776; Arenas-Huertero et al., 2000, Genes Dev. 14:2085-2096). Both the GA and ABA pathways are affected by okadaic acid, a protein phosphatase inhibitor (Kuo et al., 1996, Plant Cell. 8:259-269). The regulation of protein phosphorylation by kinases and phosphatases is accepted as a universal mechanism of cellular control (Cohen, 1992, Trends Biochem. Sci. 17:408 15 413). Likewise, the plant hormones ethylene (See, e.g., Zhou et al., 1998, Proc. NatI. Acad. Sci. USA 95:10294-10299; Beaudoin et al., 2000, Plant Cell 2000:1103-1115) and auxin (e.g. Colon-Carmona et al., 2000, Plant Physiol. 124:1728-1738) are involved in controlling plant development as well. [00111 Although several compounds are known that generally affect plant and seed 20 development, there is a clear need to specifically identify factors that are more specific for the developmental regulation of storage compound accumulation and to identify genes which have the capacity to confer altered or increased oil production to its host plant and to other plant species. This invention discloses a large number of nucleic acid sequences from Arabidopsis thaliana. These nucleic acid sequences can be used to alter 25 or increase the levels of seed storage compounds such as proteins, sugars, and oils in plants, including transgenic plants, such as rapeseed, canola, linseed, soybean, sunflower, maize, oat, rye, barley, wheat, rice, pepper, tagetes, cotton, oil palm, coconut palm, flax, castor, and peanut, which are oilseed plants containing high amounts of lipid compounds. 30 SUMMARY OF THE INVENTION [0012] The present invention provides novel isolated nucleic acid and amino acid sequences associated with the metabolism of seed storage compounds in plants. 100131 The present invention also provides isolated nucleic acids from Arabidopsis thaliana encoding a Lipid Metabolism Protein (LMP), or a portion thereof. These -5sequences may be used to modify or increase lipids and fatty acids, cofactors and enzymes in microorganisms and plants. [00141 Arabidopsis plants are known to produce considerable amounts of fatty acids like linoleic and linolenic acid (See, e.g., Table 2) and for their close similarity in many 5 aspects (gene homology, etc.) to the oil crop plant Brassica. Therefore, nucleic acid molecules originating from a plant like Arabidopsis thaliana are especially suited to modify the lipid and fatty acid metabolism in a host, especially in microorganisms and plants. Furthermore, nucleic acids from the plant Arabidopsis thaliana can be used to identify those DNA sequences and enzymes in other species, which are useful to modify 10 the biosynthesis of precursor molecules of fatty acids in the respective organisms. [0015] The present invention further provides an isolated nucleic acid comprising a fragment of at least 15 nucleotides of a nucleic acid from a plant (Arabidopsis thaliana) encoding a Lipid Metabolism Protein (LMP), or a portion thereof. 100161 Also provided by the present invention are polypeptides encoded by the 15 nucleic acids, heterologous polypeptides comprising polypeptides encoded by the nucleic acids, and antibodies to those polypeptides. [0017] Additionally, the present invention relates to and provides the use of LMP nucleic acids in the production of transgenic plants having a modified level of a seed storage compound. A method of producing a transgenic plant with a modified level of a 20 seed storage compound includes the steps of transforming a plant cell with an expression vector comprising an LMP nucleic acid, and generating a plant with a modified level of the seed storage compound from the plant cell. In one embodiment, the plant is a high oil producing species as described in Kinney et al. (1994, Current Opin. in Biotech. 5:144 151), T6pfer et al. (1995, Science 268:681-686), and Oil Crops of the World-Their 25 Breeding and Utilization (1989, eds. R~bbelen, Downey, and Ashri). In a preferred embodiment, the plant is an oil producing species selected from the group consisting of rapeseed, canola, linseed, soybean, sunflower, maize, oat, rye, barley, wheat, rice, pepper, tagetes, cotton, oil palm, coconut palm, flax, castor and peanut, for example. 100181 According to the present invention, the compositions and methods described 30 herein can be used to increase or decrease the level of an LMP in a transgenic plant comprising increasing or decreasing the expression of an LMP nucleic acid in the plant. Increased or decreased expression of the LMP nucleic acid can be achieved through transgenic expression, cosuppression, antisense inhibition, or in vivo mutagenesis of the LMP nucleic acid. The present invention also can be used to increase or decrease the -6level of a lipid in a seed oil, to increase or decrease the level of a fatty acid in a seed oil, or to increase or decrease the level of a starch in a seed or plant. [00191 The present invention provides transgenic plants having modified levels of seed storage compounds, and in particular, modified levels of a lipid, a fatty acid, or a 5 sugar. Also included herein is a seed produced by a transgenic plant transformed by an LMP DNA sequence, wherein the seed contains the LMP DNA sequence and wherein the plant is true breeding for a modified level of a seed storage compound. The present invention additionally includes a seed oil produced by the aforementioned seed. [00201 Further provided by the present invention are vectors comprising the nucleic 10 acids, host cells containing the vectors, and descendent plant materials produced by transforming a plant cell with the nucleic acids and/or vectors. [0021] According to the present invention, the compounds, compositions, and methods described herein can be used to increase or decrease the level of a lipid in a seed oil, to increase or decrease the level of a fatty acid in a seed oil, or to increase or decrease 15 the level of a starch or other carbohydrate in a seed or plant. A method of producing a higher or lower than normal or typical level of storage compound in a transgenic plant, comprises expressing an LMP nucleic acid from Arabidopsis thaliana in the transgenic plant, wherein the transgenic plant is Arabidopsis thaliana or a species different from Arabidopsis thaliana. Also included herein are compositions and methods of the 20 modification of the efficiency of production of a seed storage compound. [00221 The present invention provides novel isolated LMP nucleic acids and isolated LMP amino acid sequences from Arabidopsis thaliana as well as active fragments, analogs and orthologs thereof. The polynucleotides and polypeptides of the present invention, including agonists and/or fragments thereof, may have uses that include 25 modulating plant growth, and potentially plant yield, preferably increasing plant growth under adverse conditions (drought, cold, light, UV). In addition, antagonists of the present invention may have uses that include modulating plant growth and/or yield, through preferably increasing plant growth and yield. In yet another embodiment, overexpression polypeptides of the present invention using a constitutive promoter (e.g., 30 35S, or other promoters) may be useful for increasing plant yield under stress conditions (drought, light, cold, UV) by modulating light utilization efficiency. [0023] The present invention also provides methods for producing such aforementioned transgenic plants. -7- [00241 The present invention further provides seeds and seed oils from such aforementioned transgenic plants. 100251 These and other features and advantages of the present invention will become apparent after a review of the following detailed description of the disclosed 5 embodiments and the appended claims. DETAILED DESCRIPTION OF THE INVENTION 100261 The present invention may be understood more readily by reference to the following detailed description of the preferred embodiments of the invention and the 10 Examples included therein. [00271 Before the present compounds, compositions, and methods are disclosed and described, it is to be understood that this invention is not limited to specific nucleic acids, specific polypeptides, specific cell types, specific host cells, specific conditions, or specific methods, etc., as such may, of course, vary, and the numerous modifications and 15 variations therein will be apparent to those skilled in the art. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting. As used in the specification and in the claims, "a" or "an" can mean one or more, depending upon the context in which it is used. Thus, for example, reference to "a cell" can mean that at least one cell can be utilized. 20 100281 In accordance with the purposes of this invention, as embodied and described herein, this invention, in one aspect, provides an isolated nucleic acid from a plant (Arabidopsis thaliana) encoding a Lipid Metabolism Protein (LMP), or a portion thereof. 100291 One aspect of the invention pertains to isolated nucleic acid molecules that encode LMP polypeptides or biologically active portions thereof, as well as nucleic acid 25 fragments sufficient for use as hybridization probes or primers for the identification or amplification of an LMP-encoding nucleic acid (e.g., LMP DNA). As used herein, the term "nucleic acid molecule" is intended to include DNA molecules (e.g., cDNA or genomic DNA) and RNA molecules (e.g., mRNA) and analogs of the DNA or RNA generated using nucleotide analogs. This term also encompasses untranslated sequence 30 located at both the 3' and 5' ends of the coding region of a gene: at least about 1000 nucleotides of sequence upstream from the 5' end of the coding region and at least about 200 nucleotides of sequence downstream from the 3' end of the coding region of the gene. The nucleic acid molecule can be single-stranded or double-stranded, but preferably is double-stranded DNA. An "isolated" nucleic acid molecule is one which is -8substantially separated from other nucleic acid molecules which are present in the natural source of the nucleic acid. Preferably, an "isolated" nucleic acid is substantially free of sequences that naturally flank the nucleic acid (i.e., sequences located at the 5' and 3' ends of the nucleic acid) in the genomic DNA of the organism from which the nucleic 5 acid is derived. For example, in various embodiments, the isolated LMP nucleic acid molecule can contain less than about 5 kb, 4kb, 3kb, 2kb, 1 kb, 0.5 kb or 0.1 kb of nucleotide sequences which naturally flank the nucleic acid molecule in genomic DNA of the cell from which the nucleic acid is derived (e.g., an Arabidopsis thaliana cell). Moreover, an "isolated" nucleic acid molecule, such as a cDNA molecule, can be 10 substantially free of other cellular material, or culture medium when produced by recombinant techniques, or chemical precursors or other chemicals when chemically synthesized. 100301 A nucleic acid molecule of the present invention, e.g., a nucleic acid molecule having a nucleotide sequence as shown in the Appendix, or a portion thereof, can be 15 isolated using standard molecular biology techniques and the sequence information provided herein. For example, an Arabidopsis thaliana LMP clDNA can be isolated from an Arabidopsis thaliana library using all or portion of one of the sequences as shown in the Appendix as a hybridization probe and standard hybridization techniques (e.g., as described in Sambrook et al. 1989, Molecular Cloning: A Laboratory Manual. 2nd, ed., 20 Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY). Moreover, a nucleic acid molecule encompassing all or a portion of one of the sequences as shown in the Appendix can be isolated by the polymerase chain reaction using oligonucleotide primers designed based upon this sequence (e.g., a nucleic acid molecule encompassing all or a portion of one of the sequences as shown in the 25 Appendix can be isolated by the polymerase chain reaction using oligonucleotide primers designed based upon this same sequence as shown in the Appendix). For example, mRNA can be isolated from plant cells (e.g., by the guanidinium-thiocyanate extraction procedure of Chirgwin et al., 1979, Biochemistry 18:5294-5299) and cDNA can be prepared using reverse transcriptase (e.g., Moloney MLV reverse transcriptase, available 30 from Gibco/BRL, Bethesda, MD; or AMV reverse transcriptase, available from Seikagaku America, Inc., St. Petersburg, FL). Synthetic oligonucleotide primers for polymerase chain reaction amplification can be designed based upon one of the nucleotide sequences as shown in the Appendix. A nucleic acid of the invention can be amplified using cDNA or, alternatively, genomic DNA, as a template and appropriate -9oligonucleotide primers according to standard PCR amplification techniques. The nucleic acid so amplified can be cloned into an appropriate vector and characterized by DNA sequence analysis. Furthermore, oligonucleotides corresponding to an LMP nucleotide sequence can be prepared by standard synthetic techniques, e.g., using an 5 automated DNA synthesizer. 10031] In a preferred embodiment, an isolated nucleic acid of the invention comprises one of the nucleotide sequences as shown in the Appendix. The sequences as shown in the Appendix correspond to the Arabidopsis thaliana LMP cDNAs of the invention. These cDNAs comprise sequences encoding LMPs (i.e., the "coding region", as shown in 10 the Appendix), as well as 5' untranslated sequences and 3' untranslated sequences. Alternatively, the nucleic acid molecules can comprise only the coding region of any of the sequences as shown in the Appendix or can contain whole genomic fragments isolated from genomic DNA. [00321 For the purposes of this application, it will be understood that each of the 15 sequences set forth in the Appendix has an identifying entry number (e.g., pk309). Each of these sequences may generally comprise three parts: a 5' upstream region, a coding region, and a downstream region. A coding region of these sequences is indicated as "ORF position" (Table 3). [00331 In another preferred embodiment, an isolated nucleic acid molecule of the 20 invention comprises a nucleic acid molecule, which is a complement of one of the nucleotide sequences shown as shown in the Appendix, or a portion thereof. A nucleic acid molecule which is complementary to one of the nucleotide sequences shown as shown in the Appendix is one which is sufficiently complementary to one of the nucleotide sequences shown as shown in the Appendix such that it can hybridize to one 25 of the nucleotide sequences shown as shown in the Appendix, thereby forming a stable duplex. [00341 In still another preferred embodiment, an isolated nucleic acid molecule of the invention comprises a nucleotide sequence which is at least about 50-60%, preferably at least about 60-70%, more preferably at least about 70-80%, 80-90%, or 90-95%, and 30 even more preferably at least about 95%, 96%, 97%, 98%, 99% or more homologous to a nucleotide sequence shown as shown in the Appendix, or a portion thereof. In an additional preferred embodiment, an isolated nucleic acid molecule of the invention comprises a nucleotide sequence which hybridizes, e.g., hybridizes under stringent conditions, to one of the nucleotide sequences shown as shown in the Appendix, or a -10portion thereof. These hybridization conditions include washing with a solution having a salt concentration of about 0.02 M at pH 7 at about 600C. [0035] Moreover, the nucleic acid molecule of the invention can comprise only a portion of the coding region of one of the sequences shown in the Appendix, for example, 5 a fragment, which can be used as a probe or primer or a fragment encoding a biologically active portion of an LMP. The nucleotide sequences determined from the cloning of the LMP genes from Arabidopsis thaliana allows for the generation of probes and primers designed for use in identifying and/or cloning LMP homologues in other cell types and organisms, as well as LMP homologues from other plants or related species. Therefore 10 this invention also provides compounds comprising the nucleic acids disclosed herein, or fragments thereof. These compounds include the nucleic acids attached to a moiety. These moieties include, but are not limited to, detection moieties, hybridization moieties, purification moieties, delivery moieties, reaction moieties, binding moieties, and the like. The probe/primer typically comprises substantially purified oligonucleotide. The 15 oligonucleotide typically comprises a region of nucleotide sequence that hybridizes under stringent conditions to at least about 12, preferably about 25, more preferably about 40, 50, or 75 consecutive nucleotides of a sense strand of one of the sequences set forth in the Appendix, an anti-sense sequence of one of the sequences set forth in the Appendix, or naturally occurring mutants thereof. Primers based on a nucleotide sequence as shown in 20 the Appendix can be used in PCR reactions to clone LMP homologues. Probes based on the LMP nucleotide sequences can be used to detect transcripts or genomic sequences encoding the same or homologous proteins. In preferred embodiments, the probe further comprises a label group attached thereto, e.g. the label group can be a radioisotope, a fluorescent compound, an enzyme, or an enzyme co-factor. Such probes can be used as a 25 part of a genomic marker test kit for identifying cells which express an LMP, such as by measuring a level of an LMP-encoding nucleic acid in a sample of cells, e.g., detecting LMP mRNA levels or determining whether a genomic LMIP gene has been mutated or deleted. 10036] In one embodiment, the nucleic acid molecule of the invention encodes a 30 protein or portion thereof which includes an amino acid sequence which is sufficiently homologous to an amino acid encoded by a sequence as shown in the Appendix such that the protein or portion thereof maintains the same or a similar function as the wild-type protein. As used herein, the language "sufficiently homologous" refers to proteins or -11portions thereof which have amino acid sequences which include a minimum number of identical or equivalent (e.g., an amino acid residue which has a similar side chain as an amino acid residue in one of the polypeptides encoded by the ORF of a sequence shown in the Appendix) amino acid residues to an amino acid sequence such that the protein or 5 portion thereof is able to participate in the metabolism of compounds necessary for the production of seed storage compounds in plants, construction of cellular membranes in microorganisms or plants, or in the transport of molecules across these membranes. Regulatory proteins, such as DNA binding proteins, transcription factors, kinases, phosphatases, or protein members of metabolic pathways such as the lipid, starch and 10 protein biosynthetic pathways, or membrane transport systems, may play a role in the biosynthesis of seed storage compounds. Examples of such activities are described herein (see putative annotations in Table 3). Examples of LMP-encoding nucleic acid sequences are set forth in the Appendix. 100371 As altered or increased sugar and/or fatty acid production is a general trait 15 wished to be inherited into a wide variety of plants like maize, wheat, rye, oat, triticale, rice, barley, soybean, peanut, cotton, rapeseed, canola, manihot, pepper, sunflower, sugarbeet, tagetes, solanaceous plants like potato, tobacco, eggplant, and tomato, Vicia species, pea, alfalfa, bushy plants (coffee, cacao, tea), Salix species, trees (oil palm, coconut) and perennial grasses and forage crops, these crop plants are also preferred 20 target plants for genetic engineering as one further embodiment of the present invention. [00381 Portions of proteins encoded by the LMP nucleic acid molecules of the invention are preferably biologically active portions of one of the LMPs. As used herein, the term "biologically active portion of an LMP" is intended to include a portion, e.g., a domain/motif, of an LMP that participates in the metabolism of compounds necessary for 25 the biosynthesis of seed storage lipids, or the construction of cellular membranes in microorganisms or plants, or in the transport of molecules across these membranes, or has an activity as set forth in Table 3. To determine whether an LMP or a biologically active portion thereof can participate in the metabolism of compounds necessary for the production of seed storage compounds and cellular membranes, an assay of enzymatic 30 activity may be performed. Such assay methods are well known to those skilled in the art, and as described in Example 14. 100391 Biologically active portions of an LMP include peptides comprising amino acid sequences derived from the amino acid sequence of an LMP (e.g., an amino acid sequence encoded by a nucleic acid as shown in the Appendix or the amino acid -12sequence of a protein homologous to an LMP, which include fewer amino acids than a full length LMP or the full length protein which is homologous to an LMP) and exhibit at least one activity of an LMP. Typically, biologically active portions (peptides, e.g., peptides which are, for example, 5, 10, 15, 20, 30, 35, 36, 37, 38, 39, 40, 50, 100, or more 5 amino acids in length) comprise a domain or motif with at least one activity of an LMP. Moreover, other biologically active portions, in which other regions of the protein are deleted, can be prepared by recombinant techniques and evaluated for one or more of the activities described herein. Preferably, the biologically active portions of an LMP include one or more selected domains/motifs or portions thereof having biological 10 activity. [0040] Additional nucleic acid fragments encoding biologically active portions of an LMP can be prepared by isolating a portion of one of the sequences, expressing the encoded portion of the LMP or peptide (e.g., by recombinant expression in vitro) and assessing the activity of the encoded portion of the LMP or peptide. 15 [0041] The invention further encompasses nucleic acid molecules that differ from one of the nucleotide sequences shown as shown in the Appendix (and portions thereof) due to degeneracy of the genetic code and thus encode the same LMP as that encoded by the nucleotide sequences shown as shown in the Appendix. In a further embodiment, the nucleic acid molecule of the invention encodes a full length protein which is substantially 20 homologous to an amino acid sequence of a polypeptide encoded by an open reading frame shown as shown in the Appendix. In one embodiment, the full-length nucleic acid or protein or fragment of the nucleic acid or protein is from Arabidopsis thaliana. [00421 In addition to the Arabidopsis thaliana LMP nucleotide sequences shown as shown in the Appendix, it will be appreciated by those skilled in the art that DNA 25 sequence polymorphisms that lead to changes in the amino acid sequences of LM[Ps may exist within a population (e.g., the Arabidopsis thaliana population). Such genetic polymorphism in the LMP gene may exist among individuals within a population due to natural variation. As used herein, the terms "gene" and "recombinant gene" refer to nucleic acid molecules comprising an open reading frame encoding an LMP, preferably a 30 Arabidopsis thaliana LMP. Such natural variations can typically result in 1-40% variance in the nucleotide sequence of the LMP gene. Any and all such nucleotide variations and resulting amino acid polymorphisms in LMP that are the result of natural variation and that do not alter the functional activity of LMPs are intended to be within the scope of the invention. -13- 100431 Nucleic acid molecules corresponding to natural variants and non-Arabidopsis thaliana orthologs of the Arabidopsis thaliana LMP cDNA of the invention can be isolated based on their homology to Arabidopsis thaliana LMP nucleic acid disclosed herein using the Arabidopsis thaliana cDNA, or a portion thereof, as a hybridization 5 probe according to standard hybridization techniques under stringent hybridization conditions.. As used herein, the term "orthologs" refers to two nucleic acids from different species, but that have evolved from a common ancestral gene by speciation. Normally, orthologs encode proteins having the same or similar functions. Accordingly, in another embodiment, an isolated nucleic acid molecule of the invention is at least 15 10 nucleotides in length and hybridizes under stringent conditions to the nucleic acid molecule comprising a nucleotide sequence as shown in the Appendix. In other embodiments, the nucleic acid is at least 30, 50, 100, 250, or more nucleotides in length. As used herein, the term "hybridizes under stringent conditions" is intended to describe conditions for hybridization and washing under which nucleotide sequences at least 60% 15 homologous to each other typically remain hybridized to each other. Preferably, the conditions are such that sequences at least about 65%, more preferably at least about 70%, and even more preferably at least about 75% or more homologous to each other typically remain hybridized to each other. Such stringent conditions are known to those skilled in the art and can be found in Current Protocols in Molecular Biology, John Wiley 20 & Sons, N.Y., 1989, 6.3.1-6.3.6. A preferred, non-limiting example of stringent hybridization conditions are hybridization in 6X sodium chloride/sodium citrate (SSC) at about 45*C, followed by one or more washes in 0.2 X SSC, 0.1% SDS at 50-65*C. Another preferred example of stringent hybridization conditions is hybridization in a 6X SSC solution at 65*C. Preferably, an isolated nucleic acid molecule of the invention that 25 hybridizes under stringent conditions to a sequence as shown in the Appendix corresponds to a naturally occurring nucleic acid molecule. As used herein, a "naturally occurring" nucleic acid molecule refers to an RNA or DNA molecule having a nucleotide sequence that occurs in nature (e.g., encodes a natural protein). In one embodiment, the nucleic acid encodes a naturally occurring Arabidopsis thaliana LMP. 30 [00441 In addition to naturally-occurring variants of the LMP sequence that may exist in the population, the skilled artisan will further appreciate that changes can be introduced by mutation into a nucleotide sequence as shown in the Appendix, thereby leading to changes in the amino acid sequence of the encoded LMP, without altering the functional ability of the LMP. For example, nucleotide substitutions leading to amino -14acid substitutions at "non-essential" amino acid residues can be made in a sequence as shown in the Appendix. A "non-essential" amino acid residue is a residue that can be altered from the wild-type sequence of one of the LMPs (polypeptides encoded by any of the sequences as shown in the Appendix) without altering the activity of said LMP, 5 whereas an "essential" amino acid residue is required for LMP activity. Other amino acid residues, however, (e.g., those that are not conserved or only semi-conserved in the domain having LMP activity) may not be essential for activity and thus are likely to be amenable to alteration without altering LMP activity. 100451 Accordingly, another aspect of the invention pertains to nucleic acid 10 molecules encoding LMPs that contain changes in amino acid residues that are not essential for LMP activity. Such LMPs differ in amino acid sequence from a sequence yet retain at least one of the LMP activities described herein. In one embodiment, the isolated nucleic acid molecule comprises a nucleotide sequence encoding a protein, wherein the protein comprises an amino acid sequence at least about 50% homologous to 15 an amino acid sequence encoded by a nucleic acid as shown in the Appendix and is capable of participation in the metabolism of compounds necessary for the production of seed storage compounds in Arabidopsis thaliana, or cellular membranes, or has one or more activities set forth in Table 3. Preferably, the protein encoded by the nucleic acid molecule is at least about 50-60% homologous to one of the sequences encoded by a 20 nucleic acid as shown in the Appendix, more preferably at least about 60-70% homologous to one of the sequences encoded by a nucleic acid as shown in the Appendix, even more preferably at least about 70-80%, 80-90%, 90-95% homologous to one of the sequences encoded by a nucleic acid as shown in the Appendix, and most preferably at least about 96%, 97%, 98%, or 99% homologous to one of the sequences encoded by a 25 nucleic acid as shown in the Appendix. [0046] To determine the percent homology of two amino acid sequences (e.g., one of the sequences encoded by a nucleic acid as shown in the Appendix and a mutant form thereof) or of two nucleic acids, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of one protein or nucleic acid for 30 optimal alignment with the other protein or nucleic acid). The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in one sequence (e.g., one of the sequences encoded by a nucleic acid as shown in the Appendix) is occupied by the same amino acid residue or nucleotide as the corresponding position in the other sequence (e.g., a mutant form of the -15sequence selected from the polypeptide encoded by a nucleic acid as shown in the Appendix), then the molecules are homologous at that position (i.e., as used herein amino acid or nucleic acid "homology" is equivalent to amino acid or nucleic acid "identity"). The percent homology between the two sequences is a function of the number of identical 5 positions shared by the sequences (i.e., % homology = numbers of identical positions/total numbers of positions x 100). 10047] An isolated nucleic acid molecule encoding an LMP homologous to a protein sequence encoded by a nucleic acid as shown in the Appendix can be created by introducing one or more nucleotide substitutions, additions or deletions into a nucleotide 10 sequence as shown in the Appendix such that one or more amino acid substitutions, additions or deletions are introduced into the encoded protein. Mutations can be introduced into one of the sequences as shown in the Appendix by standard techniques, such as site-directed mutagenesis and PCR-mediated mutagenesis. Preferably, conservative amino acid substitutions are made at one or more predicted non-essential 15 amino acid residues. A "conservative amino acid substitution" is one in which the amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., 20 glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). Thus, a predicted non-essential amino acid residue in an LMP is preferably replaced with another amino acid residue 25 from the same side chain family. Alternatively, in another embodiment, mutations can be introduced randomly along all or part of an LMP coding sequence, such as by saturation mutagenesis, and the resultant mutants can be screened for an LMP activity described herein to identify mutants that retain LMP activity. Following mutagenesis of one of the sequences as shown in the Appendix, the encoded protein can be expressed 30 recombinantly, and the activity of the protein can be determined using, for example, assays described herein (See Examples 11-13). [00481 LMPs are preferably produced by recombinant DNA techniques. For example, a nucleic acid molecule encoding the protein is cloned into an expression vector (as described above), the expression vector is introduced into a host cell (as described -16herein) and the LMP is expressed in the host cell. The LMP can then be isolated from the cells by an appropriate purification scheme using standard protein purification techniques. Alternative to recombinant expression, an LMP or peptide thereof can be synthesized chemically using standard peptide synthesis techniques. Moreover, native 5 LMP can be isolated from cells, for example using an anti-LMP antibody, which can be produced by standard techniques utilizing an LMP or fragment thereof of this invention. 10049] The invention also provides LMP chimeric or fusion proteins. As used herein, an LMP "chimeric protein" or "fusion protein" comprises an LMP polypeptide operatively linked to a non-LMP polypeptide. An "LMP polypeptide" or "LMP protein" 10 refers to a polypeptide having an amino acid sequence corresponding to an LMP, whereas a "non-LMP polypeptide" refers to a polypeptide having an amino acid sequence corresponding to a protein which is not substantially homologous to the LMP, e.g., a protein which is different from the LMP and which is derived from the same or a different organism. With respect to the fusion protein, the term "operatively linked" is 15 intended to indicate that the LMP polypeptide and the non-LMP polypeptide are fused to each other so that both sequences fulfill the proposed function attributed to the sequence used. The non-LMP polypeptide can be fused to the N-terminus or C-terminus of the LMP polypeptide. For example, in one embodiment, the fusion protein is a GST-LMP (glutathione S-transferase) fusion protein in which the LMP sequences are fused to the C 20 terminus of the GST sequences. Such fusion proteins can facilitate the purification of recombinant LMPs. In another embodiment, the fusion protein is an LMP containing a heterologous signal sequence at its N-terminus. In certain host cells (e.g., mammalian host cells), expression and/or secretion of an LMP can be increased through use of a heterologous signal sequence. 25 [00501 Preferably, an LMP chimeric or fusion protein of the invention is produced by standard recombinant DNA techniques. For example, DNA fragments coding for the different polypeptide sequences are ligated together in-frame in accordance with conventional techniques, for example by employing blunt-ended or stagger-ended termini for ligation, restriction enzyme digestion to provide for appropriate termini, filling-in of 30 cohesive ends as appropriate, alkaline phosphatase treatment to avoid undesirable joining, and enzymatic ligation. In another embodiment, the fusion gene can be synthesized by conventional techniques including automated DNA synthesizers. Alternatively, PCR amplification of gene fragments can be carried out using anchor primers that give rise to complementary overhangs between two consecutive gene fragments, which can -17subsequently be annealed and reamplified to generate a chimeric gene sequence (See, for example, Current Protocols in Molecular Biology, eds. Ausubel et al., John Wiley & Sons: 1992). Moreover, many expression vectors are commercially available that already encode a fusion moiety (e.g., a GST polypeptide). An LMP-encoding nucleic acid can be 5 cloned into such an expression vector such that the fusion moiety is linked in-frame to the LMP. 100511 In addition to the nucleic acid molecules encoding LMPs described above, another aspect of the invention pertains to isolated nucleic acid molecules that are antisense thereto. An "antisense" nucleic acid comprises a nucleotide sequence that is 10 complementary to a "sense" nucleic acid encoding a protein, e.g., complementary to the coding strand of a double-stranded cDNA molecule or complementary to an mRNA sequence. Accordingly, an antisense nucleic acid can hydrogen bond to a sense nucleic acid. The antisense nucleic acid can be complementary to an entire LMP coding strand, or to only a portion thereof. In one embodiment, an antisense nucleic acid molecule is 15 antisense to a "coding region" of the coding strand of a nucleotide sequence encoding an LMP. The term "coding region" refers to the region of the nucleotide sequence comprising codons that are translated into amino acid residues (e.g., the entire coding region of pk309 comprises nucleotides 214 to 1299). In another embodiment, the antisense nucleic acid molecule is antisense to a "noncoding region" of the coding strand 20 of a nucleotide sequence encoding LMP. The term "noncoding region" refers to 5' and 3' sequences which flank the coding region, that are not translated into amino acids (i.e., also referred to as 5' and 3' untranslated regions). 10052] Given the coding strand sequences encoding LMP disclosed herein (e.g., the sequences as shown in the Appendix), antisense nucleic acids of the invention can be 25 designed according to the rules of Watson and Crick base pairing. The antisense nucleic acid molecule can be complementary to the entire coding region of LMP mRNA, but more preferably is an oligonucleotide. which is antisense to only a portion of the coding or noncoding region of LMP mRNA. For example, the antisense oligonucleotide can be complementary to the region surrounding the translation start site of LMP mRNA. An 30 antisense oligonucleotide can be, for example, about 5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 nucleotides in length. An antisense or sense nucleic acid of the invention can be constructed using chemical synthesis and enzymatic ligation reactions using procedures known in the art. For example, an antisense nucleic acid (e.g., an antisense oligonucleotide) can be chemically synthesized using naturally occurring nucleotides or -18variously modified nucleotides designed to increase the biological stability of the molecules or to increase the physical stability of the duplex formed between the antisense and sense nucleic acids, e.g., phosphorothioate derivatives and acridine substituted nucleotides can be used. Examples of modified nucleotides which can be used to 5 generate the antisense nucleic acid include 5-fluorouracil, 5-bromouracil, 5-chlorouracil, 5-iodouracil, hypoxanthine, xanthine, 4-acetylcytosine, 5-(carboxyhydroxylmethyl) uracil, 5-carboxymethylamino-methyl-2-thiouridine, 5-carboxymethylaminomethyluracil, dihydrouracil, beta-D-galactosylqueosine, inosine, N-6-isopentenyladenine, I methylguanine, 1-methylinosine, 2,2-dimethylguanine, 2-methyladenine, 2-methyl 10 guanine, 3-methylcytosine, 5-methyl-cytosine, N-6-adenine, 7-methylguanine, 5-methyl aminomethyluracil, 5-methoxyamino-methyl-2-thiouracil, beta-D-mannosylqueosine, 5' methoxycarboxymethyl-uracil, 5-methoxyuracil, 2-methylthio-N-6-isopentenyladenine, uracil-5-oxyacetic acid (v), wybutoxosine, pseudouracil, queosine, 2-thiocytosine, 5 methyl-2-thiouracil, 2-thiouracil, 4-thiouracil, 5-methyluracil, uracil-5- oxyacetic acid 15 methylester, uracil-5-oxyacetic acid (v), 5-methyl-2-thiouracil, 3-(3-amino-3-N-2 carboxypropyl) uracil, (acp3)w, and 2,6-diamino-purine. Alternatively, the antisense nucleic acid can be produced biologically using an expression vector into which a nucleic acid has been subcloned in an antisense orientation (i.e., RNA transcribed from the inserted nucleic acid will be of an antisense orientation to a target nucleic acid of interest, 20 described further in the following subsection). 100531 In another variation of the antisense technology, a double-strand interfering RNA construct can be used to cause a down-regulation of the LMP mRNA level and LMP activity in transgenic plants. This requires transforming the plants with a chimeric construct containing a portion of the LMP sequence in the sense orientation fused to the 25 antisense sequence of the same portion of the LMP sequence. A DNA linker region of variable length can be used to separate the sense and antiscnse fragments of LMP sequences in the construct. [00541 The antisense nucleic acid molecules of the invention are typically administered to a cell or generated in situ such that they hybridize with or bind to cellular 30 mRNA and/or genomic DNA encoding an LMP to thereby inhibit expression of the protein, e.g., by inhibiting transcription and/or translation. The hybridization can be by conventional nucleotide complementarity to form a stable duplex, or, for example, in the case of an antisense nucleic acid molecule which binds to DNA duplexes, through specific interactions in the major groove of the double helix. The antisense molecule can -19be modified such that it specifically binds to a receptor or an antigen expressed on a selected cell surface, e.g., by linking the antisense nucleic acid molecule to a peptide or an antibody which binds to a cell surface receptor or antigen. The antisense nucleic acid molecule can also be delivered to cells using the vectors described herein. To achieve 5 sufficient intracellular concentrations of the antisense molecules, vector constructs in which the antisense nucleic acid molecule is placed under the control of a strong prokaryotic, viral, or eukaryotic including plant promoters are preferred. [00551 In yet another embodiment, the antisense nucleic acid molecule of the invention is an at-anomeric nucleic acid molecule. An t-anomeric nucleic acid molecule 10 forms specific double-stranded hybrids with complementary RNA in which, contrary to the usual units, the strands run parallel to each other (Gaultier et al., 1987, Nucleic Acids Res. 15:6625-6641). The antisense nucleic acid molecule can also comprise a 2'-o methyl-ribonucleotide (Inoue et al., 1987, Nucleic Acids Res. 15:6131-6148) or a chimeric RNA-DNA analogue (Inoue et al., 1987, FEBS Lett. 215:327-330). 15 100561 In still another embodiment, an antisense nucleic acid of the invention is a ribozyme. Ribozymes are catalytic RNA molecules with ribonuclease activity, which are capable of cleaving a single-stranded nucleic acid, such as an mRNA, to which they have a complementary region. Thus, ribozymes (e.g., hammerhead ribozymes (described in Haselboff & Gerlach, 1988, Nature 334:585-591)) can be used to catalytically cleave 20 LMP mRNA transcripts to thereby inhibit translation of LMP mRNA. A ribozyme having specificity for an LMP-encoding nucleic acid can be designed based upon the nucleotide sequence of an LMP cDNA disclosed herein (i.e., any of the sequences as shown in the Appendix) or on the basis of a heterologous sequence to be isolated according to methods taught in this invention. For example, a derivative of a 25 Tetrahymena L-19 IVS RNA can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved in an LMP encoding mRNA (See, e.g., Cech et al., U.S. Patent No. 4,987,071 and Cech et al., U.S. Patent No. 5,116,742). Alternatively, LMP mRNA can be used to select a catalytic RNA having a specific ribonuclease activity from a pool of RNA molecules (See, e.g., Bartel 30 & Szostak, 1993, Science 261:1411-1418). [00571 Alternatively, LMP gene expression can be inhibited by targeting nucleotide sequences complementary to the regulatory region of an LMP nucleotide sequence (e.g., an LMP promoter and/or enhancer) to form triple helical structures that prevent -20transcription of an LMP gene in target cells (See generally, Helene, 1991, Anticancer Drug Des. 6:569-84; Helene et al., 1992, Ann. N.Y. Acad. Sci. 660:27-36; and Maher, 1992, Bioassays 14:807-15). [00581 Another aspect of the invention pertains to vectors, preferably expression 5 vectors, containing a nucleic acid encoding an LMP (or a portion thereof). As used herein, the term "vector" refers to a nucleic acid molecule capable of transporting another nucleic acid to which it has been linked. One type of vector is a "plasmid", which refers to a circular double stranded DNA loop into which additional DNA segments can be ligated. Another type of vector is a viral vector, wherein additional DNA segments can 10 be ligated into the viral genome. Certain vectors are capable of autonomous replication in a host cell into which they are introduced (e.g., bacterial vectors having a bacterial origin of replication and episomal mammalian vectors). Other vectors (e.g., non episomal mammalian vectors) are integrated into the genome of a host cell upon introduction into the host cell, and thereby are replicated along with the host genome. 15 Moreover, certain vectors are capable of directing the expression of genes to which they are operatively linked. Such vectors are referred to herein as "expression vectors." In general, expression vectors of utility in recombinant DNA techniques are often in the form of plasmids. In the present specification, "plasmid" and "vector" can be used interchangeably as the plasmid is the most commonly used form of vector. However, the 20 invention is intended to include such other forms of expression vectors, such as viral vectors (e.g., replication defective retroviruses, adenoviruses and adeno-associated viruses), which serve equivalent functions. 100591 The recombinant expression vectors of the invention comprise a nucleic acid of the invention in a form suitable for expression of the nucleic acid in a host cell, which 25 means that the recombinant expression vectors include one or more regulatory sequences, selected on the basis of the host cells to be used for expression, which is operatively linked to the nucleic acid sequence to be expressed. With respect to a recombinant expression vector, "operatively linked" is intended to mean that the nucleotide sequence of interest is linked to the regulatory sequence(s) in a manner which allows for expression 30 of the nucleotide sequence and both sequences are fused to each other so that each fulfills its proposed function (e.g., in an in vitro transcription/translation system or in a host cell when the vector is introduced into the host cell). The term "regulatory sequence" is intended to include promoters, enhancers, and other expression control elements (e.g., polyadenylation signals). Such regulatory sequences are described, for example, in -21- Goeddel; Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA (1990), or see Gruber and Crosby, in: Methods in Plant Molecular Biology and Biotechnolgy, CRC Press, Boca Raton, Florida, eds.: Glick & Thompson, Chapter 7, 89-108 including the references therein. Regulatory sequences include those 5 that direct constitutive expression of a nucleotide sequence in many types of host cell and those that direct expression of the nucleotide sequence only in certain host cells or under certain conditions. [t will be appreciated by those skilled in the art that the design of the expression vector can depend on such factors as the choice of the host cell to be transformed, the level of expression of protein desired, etc. The expression vectors of the 10 invention can be introduced into host cells to thereby produce proteins or peptides, including fusion proteins or peptides, encoded by nucleic acids as described herein (e.g., LMPs, mutant forms of LMPs, fusion proteins, etc.). [0060] The recombinant expression vectors of the invention can be designed for expression of LMPs in prokaryotic or eukaryotic cells. For example, LMP genes can be 15 expressed in bacterial cells, insect cells (using baculovirus expression vectors), yeast and other fungal cells (See Romanos et al., 1992, Foreign gene expression in yeast: a review, Yeast 8:423-488; van den Hondel et al., 1991, Heterologous gene expression in filamentous fungi, in: More Gene Manipulations in Fungi, Bennet & Lasure, eds., p. 396 428:Academic Press: an Diego; and van den Hondel & Punt, 1991, Gene transfer systems 20 and vector development for filamentous fungi, in: Applied Molecular Genetics of Fungi, Peberdy et al., eds., p. 1-28, Cambridge University Press: Cambridge), algae (Falciatore et al., 1999, Marine Biotechnology 1:239-251), ciliates of the types: Holotrichia, Peritrichia, Spirotrichia, Suctoria, Tetrahymena, Paramecium, Colpidium, Glaucoma, Platyophrya, Potomacus, Pseudocohnilembus, Euplotes, Engelmaniella, and Stylonychia, 25 especially of the genus Stylonychia lemnae with vectors following a transformation method as described in WO 98/01572, and multicellular plant cells (See Schmidt & Willmitzer, 1988, Plant Cell Rep.:583-586); Plant Molecular Biology and Biotechnology, C Press, Boca Raton, Florida, chapter 6/7, S.71-119 (1993); White, Jenes et al., Techniques for Gene Transfer, in: Transgenic Plants, Vol. 1, Engineering and Utilization, 30 eds.: Kung and Wu, Academic Press 1993, 128-43; Potrykus, 1991, Annu. Rev. Plant Physiol. Plant Mol. Biol. 42:205-225 (and references cited therein), or mammalian cells. Suitable host cells are discussed further in Goeddel, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, CA 1990). Alternatively, the -22recombinant expression vector can be transcribed and translated in vitro, for example using T7 promoter regulatory sequences and T7 polymerase. 100611 Expression of proteins in prokaryotes is most often carried out with vectors containing constitutive or inducible promoters directing the expression of either fusion or 5 non-fusion proteins. Fusion vectors add a number of amino acids to a protein encoded therein, usually to the amino terminus of the recombinant protein but also to the C terminus or fused within suitable regions in the proteins. Such fusion vectors typically serve one or more of the following purposes: 1) to increase expression of recombinant protein; 2) to increase the solubility of the recombinant protein; and 3) to aid in the 10 purification of the recombinant protein by acting as a ligand in affinity purification. Often, in fusion expression vectors, a proteolytic cleavage site is introduced at the junction of the fusion moiety and the recombinant protein to enable separation of the recombinant protein from the fusion moiety subsequent to purification of the fusion protein. Such enzymes, and their cognate recognition sequences, include Factor Xa, 15 thrombin, and enterokinase. [00621 Typical fusion expression vectors include pGEX (Pharmacia Biotech Inc; Smith & Johnson 1988, Gene 67:31-40), pMAL (New England Biolabs, Beverly, MA) and pRIT5 (Pharmacia, Piscataway, NJ) which fuse glutathione S-transferase (GST), maltose E binding protein, or protein A, respectively, to the target recombinant protein. 20 In one embodiment, the coding sequence of the LMP is cloned into a pGEX expression vector to create a vector encoding a fusion protein comprising, from the N-terminus to the C-terminus, GST-thrombin cleavage site-X protein. The fusion protein can be purified by affinity chromatography using glutathione-agarose resin. Recombinant LMP unfused to GST can be recovered by cleavage of the fusion protein with thrombin. 25 100631 Examples of suitable inducible non-fusion E. coli expression vectors include pTrc (Amann et al., 1988, Gene 69:301-315) and pET I Id (Studier et al., 1990, Gene Expression Technology: Methods in Enzymology 185, Academic Press, San Diego, California 60-89). Target gene expression from the pTrc vector relies on host RNA polymerase transcription from a hybrid trp-lac fusion promoter. Target gene expression 30 from the pET lId vector relies on transcription from a T7 gn10-lac fusion promoter mediated by a coexpressed viral RNA polymerase (T7 gnl). This viral polymerase is supplied by host strains BL21(DE3) or HMS174(DE3) from a resident prophage harboring a T7 gnI gene under the transcriptional control of the lacUV 5 promoter. -23- [00641 One strategy to maximize recombinant protein expression is to express the protein in a host bacteria with an impaired capacity to proteolytically cleave the recombinant protein (Gottesman, 1990, Gene Expression Technology:Methods in Enzymology 185:119-128, Academic Press, San Diego, California). Another strategy is 5 to alter the nucleic acid sequence of the nucleic acid to be inserted into an expression vector so that the individual codons for each amino acid are those preferentially utilized in the bacterium chosen for expression (Wada et al., 1992, Nucleic Acids Res. 20:2111 2118). Such alteration of nucleic acid sequences of the invention can be carried out by standard DNA synthesis techniques. 10 [00651 In another embodiment, the LMP expression vector is a yeast expression vector. Examples of vectors for expression in yeast S. cerevisiae include pYepSecl (Baldari et al., 1987, Embo J. 6:229-234), pMFa (Kurjan & Herskowitz, 1982, Cell 30:933-943), pJRY88 (Schultz et al., 1987, Gene 54:113-123), and pYES2 (Invitrogen Corporation, San Diego, CA). Vectors and methods for the construction of vectors 15 appropriate for use in other fungi, such as the filamentous fungi, include those detailed in: van den Hondel & Punt, 1991, "Gene transfer systems and vector development for filamentous fungi," in: Applied Molecular Genetics of Fungi, Peberdy et al., eds., p. 1-28, Cambridge University Press: Cambridge. [0066] Alternatively, the LMPs of the invention can be expressed in insect cells using 20 baculovirus expression vectors. Baculovirus vectors available for expression of proteins in cultured insect cells (e.g., Sf 9 cells) include the pAc series (Smith et al., 1983, Mol. Cell Biol. 3:2156-2165) and the pVL series (Lucklow & Summers, 1989, Virology 170:31-39). 100671 In yet another embodiment, a nucleic acid of the invention is expressed in 25 mammalian cells using a mammalian expression vector. Examples of mammalian expression vectors include pCDM8 (Seed, 1987, Nature 329:840) and pMT2PC (Kaufman et al., 1987, EMBO J. 6:187-195). When used in mammalian cells, the expression vector's control functions are often provided by viral regulatory elements. For example, commonly used promoters are derived from polyoma, Adenovirus 2, 30 cytomegalovirus, and Simian Virus 40. For other suitable expression systems for both prokaryotic and eukaryotic cells see chapters 16 and 17 of Sambrook, Fritsh and Maniatis, Molecular Cloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 1989. -24- [00681 In another embodiment, the LMPs of the invention may be expressed in uni cellular plant cells (such as algae; see Falciatore et al., 1999, Marine Biotechnology 1:239-251, and references therein) and plant cells from higher plants (e.g., the spermatophytes, such as crop plants). Examples of plant expression vectors include those 5 detailed in: Becker et al., 1992, Plant Mol. Biol. 20:1195-1197) and Bevan, 1984, Nucleic Acids Res. 12:8711-8721; Vectors for Gene Transfer in Higher Plants; in: Transgenic Plants, Vol. 1, Engineering and Utilization, eds.: Kung und R. Wu, Academic Press, 1993, S. 15-38). [00691 A plant expression cassette preferably contains regulatory sequences capable 10 to drive gene expression in plant cells and which are operatively linked so that each sequence can fulfill its function such as termination of transcription, including polyadenylation signals. Preferred polyadenylation signals are those originating from Agrobacterium tumefaciens t-DNA such as the gene 3 known as octopine synthase of the Ti-plasmid pTiACH5 (Gielen et al., 1984, EMBO J. 3:835) or functional equivalents 15 thereof but also all other terminators functionally active in plants are suitable. [00701 As plant gene expression is very often not limited on transcriptional levels a plant expression cassette preferably contains other operatively linked sequences like translational enhancers such as the overdrive-sequence containing the 5'-untranslated leader sequence from tobacco mosaic virus enhancing the protein per RNA ratio (Gallie 20 et al., 1987, Nucleic Acids Res. 15:8693-8711). 100711 Plant gene expression has to be operatively linked to an appropriate promoter conferring gene expression in a timely, cell, or tissue specific manner. Preferred are promoters driving constitutive expression (Benfey et al., 1989, EMBO J. 8:2195-2202) like those derived from plant viruses like the 35S CAMV (Franck et al., 1980, Cell 25 21:285-294), the 19S CaMV (See also US 5,352,605 and WO 84/02913), or plant promoters like those from Rubisco small subunit described in US 4,962,028. Even more preferred are seed-specific promoters driving expression of LMP proteins during all or selected stages of seed development. Seed-specific plant promoters are known to those of ordinary skill in the art and are identified and characterized using seed-specific mRNA 30 libraries and expression profiling techniques. Seed-specific promoters include the napin gene promoter from rapeseed (US 5,608,152), the USP-promoter from Vicia faba (Baeumlein et al., 1991, Mol. Gen. Genetics 225:459-67), the oleosin-promoter from Arabidopsis (WO 98/45461), the phaseolin-promoter from Phaseolus vulgaris (US 5,504,200), the Bce4-promoter from Brassica (WO 91/13980) or the legumin B4 -25promoter (LeB4; Baeumlein et al., 1992, Plant J. 2:233-239) as well as promoters conferring seed specific expression in monocot plants like maize, barley, wheat, rye, rice etc. Suitable promoters to note are the lpt2 or lptl-gene promoter from barley (WO 95/15389 and WO 95/23230) or those described in WO 99/16890 (promoters from the 5 barley hordein-gene, the rice glutelin gene, the rice oryzin gene, the rice prolamin gene, the wheat gliadin gene, wheat glutelin gene, the maize zein gene, the oat glutelin gene, the Sorghum kasirin-gene, and the rye secalin gene). [00721 Plant gene expression can also be facilitated via an inducible promoter (For a review, see Gatz, 1997, Annu. Rev. Plant Physiol. Plant Mol. Biol. 48:89-108). 10 Chemically inducible promoters are especially suitable if gene expression is desired in a time specific manner. Examples for such promoters are a salicylic acid inducible promoter (WO 95/19443), a tetracycline inducible promoter (Gatz et al. 1992, Plant J. 2:397-404) and an ethanol inducible promoter (WO 93/21334). [00731 Promoters responding to biotic or abiotic stress conditions are also suitable 15 promoters such as the pathogen inducible PRP 1 -gene promoter (Ward et al., 1993, Plant. Mol. Biol. 22:361-366), the heat inducible hsp80-promoter from tomato (US 5,187,267), cold inducible alpha-amylase promoter from potato (WO 96/12814) or the wound inducible pinII-promoter (EP 375091). 100741 Other preferred sequences for use in plant gene expression cassettes are 20 targeting-sequences necessary to direct the gene-product in its appropriate cell compartment (For a review, see Kermode, 1996, Crit. Rev. Plant Sci. 15:285-423 and references cited therein) such as the vacuole, the nucleus, all types of plastids like amyloplasts, chloroplasts, chromoplasts, the extracellular space, mitochondria, the endoplasmic reticulum, oil bodies, peroxisomes, and other compartments of plant cells. 25 Also especially suited are promoters that confer plastid-specific gene expression, as plastids are the compartment where precursors and some end products of lipid biosynthesis are synthesized. Suitable promoters such as the viral RNA polymerase promoter are described in WO 95/16783 and WO 97/06250 and the clpP-promoter from Arabidopsis described in WO 99/46394. 30 100751 The invention further provides a recombinant expression vector comprising a DNA molecule of the invention cloned into the expression vector in an antisense orientation. That is, the DNA molecule is operatively linked to a regulatory sequence in a manner that allows for expression (by transcription of the DNA molecule) of an RNA molecule that is antisense to LMP mRNA. Regulatory sequences operatively linked to a -26nucleic acid cloned in the antisense orientation can be chosen which direct the continuous expression of the antisense RNA molecule in a variety of cell types, for instance viral promoters and/or enhancers, or regulatory sequences can be chosen which direct constitutive, tissue specific or cell type specific expression of antisense RNA. The 5 antisense expression vector can be in the form of a recombinant plasmid, phagemid, or attenuated virus in which antisense nucleic acids are produced under the control of a high efficiency regulatory region, the activity of which can be determined by the cell type into which the vector is introduced. For a discussion of the regulation of gene expression using antisense genes see Weintraub et al. (1986, Antisense RNA as a molecular tool for 10 genetic analysis, Reviews - Trends in Genetics, Vol. 1) and Mol et al. (1990, FEBS Lett. 268:427-430). 100761 Another aspect of the invention pertains to host cells into which a recombinant expression vector of the invention has been introduced. The terms "host cell" and "recombinant host cell" are used interchangeably herein. It is to be understood that such 15 terms refer not only to the particular subject cell but also to the progeny or potential progeny of such a cell. Because certain modifications may occur in succeeding generations due to either mutation or environmental influences, such progeny may not, in fact, be identical to the parent cell, but are still included within the scope of the term as used herein. A host cell can be any prokaryotic or eukaryotic cell. For example, an LMP 20 can be expressed in bacterial cells, insect cells, fungal cells, mammalian cells (such as Chinese hamster ovary cells (CHO) or COS cells), algae, ciliates or plant cells. Other suitable host cells are known to those skilled in the art. [00771 Vector DNA can be introduced into prokaryotic or eukaryotic cells via conventional transformation or transfection techniques. As used herein, the terms 25 "transformation," "transfection," "conjugation," and "transduction" are intended to refer to a variety of art-recognized techniques for introducing foreign nucleic acid (e.g., DNA) into a host cell, including calcium phosphate or calcium chloride co-precipitation, DEAE dextran-mediated transfection, lipofection, natural competence, chemical-mediated transfer, or electroporation. Suitable methods for transforming or transfecting host cells 30 including plant cells can be found in Sambrook et al. (1989, Molecular Cloning: A Laboratory Manual. 2nd, ed., Cold Spring Harbor Laboratory, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY) and other laboratory manuals such as Methods in Molecular Biology 1995, Vol. 44, Agrobacterium protocols, ed: Gartland and Davey, Humana Press, Totowa, New Jersey. -27- 100781 For stable transfection of mammalian and plant cells, it is known that, depending upon the expression vector and transfection technique used, only a small fraction of cells may integrate the foreign DNA into their genome. In order to identify and select these integrants, a gene that encodes a selectable marker (e.g., resistance to 5 antibiotics) is generally introduced into the host cells along with the gene of interest. Preferred selectable markers include those that confer resistance to drugs, such as G418, hygromycin, kanamycin, and methotrexate, or in plants that confer resistance towards an herbicide such as glyphosate or glufosinate. A nucleic acid encoding a selectable marker can be introduced into a host cell on the same vector as that encoding an LMP or can be 10 introduced on a separate vector. Cells stably transfected with the introduced nucleic acid can be identified by, for example, drug selection (e.g., cells that have incorporated the selectable marker gene will survive, while the other cells die). 10079] To create a homologous recombinant microorganism, a vector is prepared which contains at least a portion of an LMP gene into which a deletion, addition or 15 substitution has been introduced to thereby alter, e.g., functionally disrupt, the LMP gene. Preferably, this LMP gene is an Arabidopsis thaliana LMP gene, but it can be a homologue from a related plant or even from a mammalian, yeast, or insect source. In a preferred embodiment, the vector is designed such that, upon homologous recombination, the endogenous LMP gene is functionally disrupted (i.e., no longer encodes a functional 20 protein; also referred to as a knock-out vector). Alternatively, the vector can be designed such that, upon homologous recombination, the endogenous LMP gene is mutated or otherwise altered but still encodes functional protein (e.g., the upstream regulatory region can be altered to thereby alter the expression of the endogenous LMP). To create a point mutation via homologous recombination, DNA-RNA hybrids can be used in a technique 25 known as chimeraplasty (Cole-Strauss et al., 1999, Nucleic Acids Res. 27:1323-1330 and Kmiec, 1999, American Scientist 87:240-247). Homologous recombination procedures in Arabidopsis thaliana and other crops are also well known in the art and are contemplated for use herein. [00801 In a homologous recombination vector, the altered portion of the LMP gene is 30 flanked at its 5' and 3' ends by additional nucleic acid of the LMP gene to allow for homologous recombination to occur between the exogenous LMP gene carried by the vector and an endogenous LMP gene in a microorganism or plant. The additional flanking LMP nucleic acid is of sufficient length for successful homologous recombination with the endogenous gene. Typically, several hundreds of base pairs up to -28kilobases of flanking DNA (both at the 5' and 3' ends) are included in the vector (See e.g., Thomas & Capecchi, 1987, Cell 51:503, for a description of homologous recombination vectors). The vector is introduced into a microorganism or plant cell (e.g., via polyethyleneglycol mediated DNA). Cells in which the introduced LMP gene has 5 homologously recombined with the endogenous LMP gene are selected using art-known techniques. [0081] In another embodiment, recombinant microorganisms can be produced which contain selected systems that allow for regulated expression of the introduced gene. For example, inclusion of an LMP gene on a vector placing it under control of the lac operon 10 permits expression of the LMP gene only in the presence of IPTG. Such regulatory systems are well known in the art. [0082] A host cell of the invention, such as a prokaryotic or eukaryotic host cell in culture can be used to produce (i.e., express) an LMP. Accordingly, the invention further provides methods for producing LMPs using the host cells of the invention. In one 15 embodiment, the method comprises culturing a host cell of the invention (into which a recombinant expression vector encoding an LMP has been introduced, or which contains a wild-type or altered LMP gene in it's genome) in a suitable medium until LMP is produced. In another embodiment, the method further comprises isolating LMPs from the medium or the host cell. 20 100831 Another aspect of the invention pertains to isolated LMPs, and biologically active portions thereof. An "isolated" or "purified" protein or biologically active portion thereof is substantially free of cellular material when produced by recombinant DNA techniques, or chemical precursors or other chemicals when chemically synthesized. The language "substantially free of cellular material" includes preparations of LMP in which 25 the protein is separated from cellular components of the cells in which it is naturally or recombinantly produced. In one embodiment, the language "substantially free of cellular material" includes preparations of LMP having less than about 30% (by dry weight) of non-LMP (also referred to herein as a "contaminating protein"), more preferably less than about 20% of non-LMP, still more preferably less than about 10% of non-LMP, and most 30 preferably less than about 5% non-LMP. When the LMP or biologically active portion thereof is recombinantly produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, more preferably less than about 10%, and most preferably less than about 5% of the volume of the protein preparation. The language "substantially free of chemical precursors or other chemicals" -29includes preparations of LMP in which the protein is separated from chemical precursors or other chemicals that are involved in the synthesis of the protein. In one embodiment, the language "substantially free of chemical precursors or other chemicals" includes preparations of LMP having less than about 30% (by dry weight) of chemical precursors 5 or non-LMP chemicals, more preferably less than about 20% chemical precursors or non LMP chemicals, still more preferably less than about 10% chemical precursors or non LMP chemicals, and most preferably less than about 5% chemical precursors or non LMP chemicals. In preferred embodiments, isolated proteins or biologically active portions thereof lack contaminating proteins from the same organism from which the 10 LMP is derived. Typically, such proteins are produced by recombinant expression of, for example, an Arabidopsis thaliana LMP in other plants than Arabidopsis thaliana or microorganisms, algae, or fungi. [0084] An isolated LMP or a portion thereof of the invention can participate in the metabolism of compounds necessary for the production of seed storage compounds in 15 Arabidopsis thaliana or another plant, or of cellular membranes, or has one or more of the activities set forth in Table 3. In preferred embodiments, the protein or portion thereof comprises an amino acid sequence which is sufficiently homologous to an amino acid sequence encoded by a nucleic acid as shown in the Appendix such that the protein or portion thereof maintains the ability to participate in the metabolism of compounds 20 necessary for the construction of cellular membranes in Arabidopsis thaliana, or in the transport of molecules across these membranes. The portion of the protein is preferably a biologically active portion as described herein. In another preferred embodiment, an LMP of the invention has an amino acid sequence encoded by a nucleic acid as shown in the Appendix. In yet another preferred embodiment, the LMP has an amino acid 25 sequence which is encoded by a nucleotide sequence that hybridizes, e.g., hybridizes under stringent conditions, to a nucleotide sequence as shown in the Appendix. In still another preferred embodiment, the LMP has an amino acid sequence which is encoded by a nucleotide sequence that is at least about 50-60%, preferably at least about 60-70%, more preferably at least about 70-80%, 80-90%, 90-95%, and even more preferably at 30 least about 96%, 97%, 98%, 99%, or more homologous to one of the amino acid sequences encoded by a nucleic acid as shown in the Appendix. The preferred LMPs of the present invention also preferably possess at least one of the LMP activities described herein. For example, a preferred LMP of the present invention includes an amino acid sequence encoded by a nucleotide sequence which hybridizes, e.g., hybridizes under -30stringent conditions, to a nucleotide sequence as shown in the Appendix, and which can participate in the metabolism of compounds necessary for the construction of cellular membranes in Arabidopsis thaliana, or in the transport of molecules across these membranes, or which has one or more of the activities set forth in Table 3. 5 [0085] In other embodiments, the LMP is substantially homologous to an amino acid sequence encoded by a nucleic acid as shown in the Appendix and retains the functional activity of the protein of one of the sequences encoded by a nucleic acid as shown in the Appendix yet differs in amino acid sequence due to natural variation or mutagenesis, as described in detail above. Accordingly, in another embodiment, the LMP is a protein 10 which comprises an amino acid sequence which is at least about 50-60%, preferably at least about 60-70%, and more preferably at least about 70-80, 80-90, 90-95%, and most preferably at least about 96%, 97%, 98%, 99%, or more homologous to an entire amino acid sequence and which has at least one of the LMP activities described herein. In another embodiment, the invention pertains to a full Arabidopsis thaliana protein, which 15 is substantially homologous to an entire amino acid sequence encoded by a nucleic acid as shown in the Appendix. [00861 Dominant negative mutations or trans-dominant suppression can be used to reduce the activity of an LMP in transgenics seeds in order to change the levels of seed storage compounds. To achieve this a mutation that abolishes the activity of the LMP is 20 created and the inactive non-functional LMP gene is overexpressed in the transgenic plant. The inactive trans-dominant LMP protein competes with the active endogenous LMP protein for substrate or interactions with other proteins and dilutes out the activity of the active LMP. In this way the biological activity of the LMP is reduced without actually modifying the expression of the endogenous LMP gene. This strategy was used 25 by Pontier et al to modulate the activity of plant transcription factors (Pontier et al., Plant J 2001, 27(6):529-38). 100871 Homologues of the LMP can be generated by mutagenesis, e.g., discrete point mutation or truncation of the LMP. As used herein, the term "homologue" refers to a variant form of the LMP that acts as an agonist or antagonist of the activity of the LMP. 30 An agonist of the LMP can retain substantially the same, or a subset, of the biological activities of the LMP. An antagonist of the LMP can inhibit one or more of the activities of the naturally occurring form of the LMP by, for example, competitively binding to a downstream or upstream member of the cell membrane component metabolic cascade -31which includes the LMP, or by binding to an LMP which mediates transport of compounds across such membranes, thereby preventing translocation from taking place. [00881 In an alternative embodiment, homologues of the LMP can be identified by screening combinatorial libraries of mutants, e.g., truncation mutants, of the LMP for 5 LMP agonist or antagonist activity. In one embodiment, a variegated library of LMP variants is generated by combinatorial mutagenesis at the nucleic acid level and is encoded by a variegated gene library. A variegated library of LMP variants can be produced by, for example, enzymatically ligating a mixture of synthetic oligonucleotides into gene sequences such that a degenerate set of potential LMP sequences is expressible 10 as individual polypeptides, or alternatively, as a set of larger fusion proteins (e.g., for phage display) containing the set of LMP sequences therein. There are a variety of methods that can be used to produce libraries of potential LMP homologues from a degenerate oligonucleotide sequence. Chemical synthesis of a degenerate gene sequence can be performed in an automatic DNA synthesizer, and the synthetic gene then ligated 15 into an appropriate expression vector. Use of a degenerate set of genes allows for the provision, in one mixture, of all of the sequences encoding the desired set of potential LMP sequences. Methods for synthesizing degenerate oligonucleotides are known in the art (See, e.g., Narang, 1983, Tetrahedron 39:3; Itakura et al., 1984, Annu. Rev. Biochem. 53:323; Itakura et al., 1984, Science 198:1056; Ike et al., 1983, Nucleic Acids Res. 20 11:477). [0089] In addition, libraries of fragments of the LMP coding sequences can be used to generate a variegated population of LMP fragments for screening and subsequent selection of homologues of an LMP. In one embodiment, a library of coding sequence fragments can be generated by treating a double stranded PCR fragment of an LMP 25 coding sequence with a nuclease under conditions wherein nicking occurs only about once per molecule, denaturing the double stranded DNA, renaturing the DNA to form double stranded DNA which can include sense/antisense pairs from different nicked products, removing single stranded portions from reformed duplexes by treatment with SI nuclease, and ligating the resulting fragment library into an expression vector. By this 30 method, an expression library can be derived which encodes N-terminal, C-terminal, and internal fragments of various sizes of the LMP. [00901 Several techniques are known in the art for screening gene products of combinatorial libraries made by point mutations or truncation, and for screening cDNA libraries for gene products having a selected property. Such techniques are adaptable for -32rapid screening of the gene libraries generated by the combinatorial mutagenesis of LMP homologues. The most widely used techniques, which are amenable to high through-put analysis, for screening large gene libraries typically include cloning the gene library into replicable expression vectors, transforming appropriate cells with the resulting library of 5 vectors, and expressing the combinatorial genes under conditions in which detection of a desired activity facilitates isolation of the vector encoding the gene whose product was detected. Recursive ensemble mutagenesis (REM), a technique that enhances the frequency of functional mutants in the libraries, can be used in combination with the screening assays to identify LMP homologues (Arkin & Yourvan, 1992, Proc. Natl. 10 Acad. Sci. USA 89:7811-7815; Delgrave et al., 1993, Protein Engineering 6:327-331). In another embodiment, cell based assays can be exploited to analyze a variegated LMP library, using methods well known in the art. 10091] The nucleic acid molecules, proteins, protein homologues, fusion proteins, primers, vectors, and host cells described herein can be used in one or more of the 15 following methods: identification of Arabidopsis thaliana and related organisms; mapping of genomes of organisms related to Arabidopsis thaliana; identification and localization of Arabidopsis thaliana sequences of interest; evolutionary studies; determination of LMP regions required for function; modulation of an LMP activity; modulation of the metabolism of one or more cell functions; modulation of the 20 transmembrane transport of one or more compounds; and modulation of seed storage compound accumulation. [00921 The plant Arabidopsis thaliana represents one member of higher (or seed) plants. It is related to other plants such as canola or soybean, which require light to drive photosynthesis and growth. Plants like Arabidopsis thaliana share a high degree of 25 homology on the DNA sequence and polypeptide level, allowing the use of heterologous screening of DNA molecules with probes evolving from other plants or organisms, thus enabling the derivation of a consensus sequence suitable for heterologous screening or functional annotation and prediction of gene functions in third species. The ability to identify such functions can therefore have significant relevance, e.g., prediction of 30 substrate specificity of enzymes. Further, these nucleic acid molecules may serve as reference points for the mapping of Arabidopsis genomes, or of genomes of related organisms. [00931 The LMP nucleic acid molecules of the invention have a variety of uses. First, they may be used to identify an organism as being Arabidopsis thaliana or a close -33relative thereof. Also, they may be used to identify the presence of Arabidopsis thaliana or a relative thereof in a mixed population of microorganisms. The invention provides the nucleic acid sequences of a number of Arabidopsis thaliana genes; by probing the extracted genomic DNA of a culture of a unique or mixed population of microorganisms 5 under stringent conditions with a probe spanning a region of an Arabidopsis thaliana gene, which is unique to this organism, one can ascertain whether this organism is present. 100941 Further, the nucleic acid and protein molecules of the invention may serve as markers for specific regions of the genome. This has utility not only in the mapping of 10 the genome, but also for functional studies of Arabidopsis thaliana proteins. For example, to identify the region of the genome to which a particular Arabidopsis thaliana DNA-binding protein binds, the Arabidopsis thaliana genome could be digested, and the fragments incubated with the DNA-binding protein. Those which bind the protein may be additionally probed with the nucleic acid molecules of the invention, preferably with 15 readily detectable labels; binding of such a nucleic acid molecule to the genome fragment enables the localization of the fragment to the genome map of Arabidopsis thaliana, and, when performed multiple times with different enzymes, facilitates a rapid determination of the nucleic acid sequence to which the protein binds. Further, the nucleic acid molecules of the invention may be sufficiently homologous to the sequences of related 20 species such that these nucleic acid molecules may serve as markers for the construction of a genomic map in related plants. [0095] The LMP nucleic acid molecules of the invention are also useful for evolutionary and protein structural studies. The metabolic and transport processes in which the molecules of the invention participate are utilized by a wide variety of 25 prokaryotic and eukaryotic cells; by comparing the sequences of the nucleic acid molecules of the present invention to those encoding similar enzymes from other organisms, the evolutionary relatedness of the organisms can be assessed. Similarly, such a comparison permits an assessment of which regions of the sequence are conserved and which are not, which may aid in determining those regions of the protein which are 30 essential for the functioning of the enzyme. This type of determination is of value for protein engineering studies and may give an indication of what the protein can tolerate in terms of mutagenesis without losing function. 100961 Manipulation of the LMP nucleic acid molecules of the invention may result in the production of LMPs having functional differences from the wild-type LMPs. -34- These proteins may be improved in efficiency or activity, may be present in greater numbers in the cell than is usual, or may be decreased in efficiency or activity. 100971 There are a number of mechanisms by which the alteration of an LMP of the invention may directly affect the accumulation and/or composition of seed storage 5 - compounds. In the case of plants expressing LMPs, increased transport can lead to altered accumulation of compounds and/or solute partitioning within the plant tissue and organs which ultimately could be used to affect the accumulation of one or more seed storage compounds during seed development. An example is provided by Mitsukawa et al. (1997, Proc. Natl. Acad. Sci. USA 94:7098-7102), where overexpression of an 10 Arabidopsis high-affinity phosphate transporter gene in tobacco cultured cells enhanced cell growth under phosphate-limited conditions. Phosphate availability also affects significantly the production of sugars and metabolic intermediates (Hurry et al., 2000, Plant J. 24:383-396) and the lipid composition in leaves and roots (Hartel et al., 2000, Proc. Nat]. Acad. Sci. USA 97:10649-10654). Likewise, the activity of the plant ACCase 15 has been demonstrated to be regulated by phosphorylation (Savage & Ohlrogge, 1999, Plant J. 18:521-527), and alterations in the activity of the kinases and phosphatases (LMPs) that act on the ACCase could lead to increased or decreased levels of seed lipid accumulation. Moreover, the presence of lipid kinase activities in chloroplast envelope membranes suggests that signal transduction pathways and/or membrane protein 20 regulation occur in envelopes (See, e.g., Miller et al., 2000, J. Biol. Chem. 275:19475 19481 and literature cited therein). The ABJI and ABI2 genes encode two protein serine/threonine phosphatases 2C, which are regulators in abscisic acid signaling pathway, and thereby in early and late seed development (e.g. Merlot et al., 2001, Plant J. 25:295-303). For more examples, see also the section 'Background of the Invention.' 25 [00981 The present invention also provides antibodies, which specifically binds to an LMP-polypeptide, or a portion thereof, as encoded by a nucleic acid disclosed herein or as described herein. Antibodies can be made by many well-known methods (See, e.g. Harlow and Lane, "Antibodies; A Laboratory Manual" Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, 1988). Briefly, purified antigen can be injected into an 30 animal in an amount and in intervals sufficient to elicit an immune response. Antibodies can either be purified directly, or spleen cells can be obtained from the animal. The cells can then fused with an immortal cell line and screened for antibody secretion. The antibodies can be used to screen nucleic acid clone libraries for cells secreting the -35antigen. Those positive clones can then be sequenced (See, for example, Kelly et al., 1992, BioTechnology 10:163-167; Bebbington et al., 1992, BioTechnology 10:169-175). 10099] The phrase "selectively binds" with the polypeptide refers to a binding reaction, which is determinative of the presence of the protein in a heterogeneous 5 population of proteins and other biologics. Thus, under designated immunoassay conditions, the specified antibodies bound to a particular protein do not bind in a significant amount to other proteins present in the sample. Selective binding to an antibody under such conditions may require an antibody that is selected for its specificity for a particular protein. A variety of immunoassay formats may be used to select 10 antibodies that selectively bind with a particular protein. For example, solid-phase ELISA immunoassays are routinely used to select antibodies selectively immunoreactive with a protein. See Harlow and Lane "Antibodies, A Laboratory Manual" Cold Spring Harbor Publications, New York (1988), for a description of immunoassay formats and conditions that could be used to determine selective binding. 15 100100] In some instances, it is desirable to prepare monoclonal antibodies from various hosts. A description of techniques for preparing such monoclonal antibodies may be found in Stites et al., editors, "Basic and Clinical Immunology," (Lange Medical Publications, Los Altos, Calif., Fourth Edition) and references cited therein, and in Harlow and Lane ("Antibodies, A Laboratory Manual" Cold Spring Harbor Publications, 20 New York, 1988). [001011 Throughout this application, various publications are referenced. The disclosures of all of these publications and those references cited within those publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art to which this invention pertains. 25 1001021 It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the scope or spirit of the invention. Other embodiments of the invention will be apparent to those skilled in the art from consideration of the specification and practice of the invention disclosed herein. It is intended that the specification and Examples be considered as exemplary 30 only, with a true scope and spirit of the invention being indicated by the claims included herein. -36- EXAMPLES Example 1 1001031 General Processes 1001041 a) General Cloning Processes: 5 1001051 Cloning processes such as, for example, restriction cleavages, agarose gel electrophoresis, purification of DNA fragments, transfer of nucleic acids to nitrocellulose and nylon membranes, linkage of DNA fragments, transformation of Escherichia coli and yeast cells, growth of bacteria and sequence analysis of recombinant DNA were carried out as described in Sambrook et al. (1989, Cold Spring Harbor Laboratory Press: ISBN 0 10 87969-309-6) or Kaiser, Michaelis and Mitchell (1994, "Methods in Yeast Genetics," Cold Spring Harbor Laboratory Press: ISBN 0-87969-451-3). 1001061 b) Chemicals: [001071 The chemicals used were obtained, if not mentioned otherwise in the text, in p.a. quality from the companies Fluka (Neu-Ulm), Merck (Darmstadt), Roth (Karlsruhe), 15 Serva (Heidelberg), and Sigma (Deisenhofen). Solutions were prepared using purified, pyrogen-free water, designated as H20 in the following text, from a Milli-Q water system water purification plant (Milliporc, Eschborn). Restriction endonucleases, DNA modifying enzymes and molecular biology kits were obtained from the companies AGS (Heidelberg), Amersham (Braunschweig), Biometra (Gbttingen), Boehringer 20 (Mannheim), Genomed (Bad Oeynnhausen), New England Biolabs (Schwalbach/ Taunus), Novagen (Madison, Wisconsin, USA), Perkin-Elmer (Weiterstadt), Pharmacia (Freiburg), Qiagen (Hilden) and Stratagene (Amsterdam, Netherlands). They were used, if not mentioned otherwise, according to the manufacturer's instructions. 1001081 c) Plant Material and Growth 25 100109] Arabidopsis thaliana plants 1001101 For this study, root material, leaves, siliques and seeds of wild-type and mutant plants of Arabidopsis thaliana were used. The wril mutation was isolated from an ethyl methanesulfonate-mutagenized population of the Columbia ecotype as described (Benning et al. 1998, Plant Physiol 118:91-101). Wild type and wril Arabidopsis seeds 30 were preincubated for three days in the dark at 4*C before placing them into an incubator (AR-75, Percival Scientific, Boone, IA) at a photon flux density of 60-80 pmol m 2 s' and a light period of 16 hours (22*C), and a dark period of 8 hours (18*C). All plants were started on half-strength MS medium (Murashige & Skoog, 1962, Plant Physiol. 15:473-497), pH 6.2, 2% sucrose and 1.2% agar. Seeds were sterilized for 20 minutes in -37- 20% bleach 0.5% triton X100 and rinsed 6 times with excess sterile water. Plants were either grown as described above or on soil under standard conditions as described in Focks & Benning (1998, Plant Physiol 118:91-101). [001111 In other series of experiments, siliques of individual ecotypes of Arabidopsis 5 thaliana and of selected Arabidopsis mutants were used. Seeds were obtained from the Arabidopsis stock center. Example 2 Total DNA Isolation from Plants 10 [001121 The details for the isolation of total DNA relate to the working up of one gram fresh weight of plant material. [001131 CTAB buffer: 2% (w/v) N-cethyl-N,N,N-trimethylammonium bromide (CTAB); 100 mM Tris HCl pH 8.0; 1.4 M NaCl; 20 mM EDTA. N-Laurylsarcosine buffer:10% (w/v) N-laurylsarcosine; 100 mM Tris HCl pH 8.0; 20 mM EDTA. 15 [001141 The plant material was triturated under liquid nitrogen in a mortar to give a fine powder and transferred to 2 ml Eppendorf vessels. The frozen plant material was then covered with a layer of 1 ml of decomposition buffer (I ml CTAB buffer, 100 pl of N-laurylsarcosine buffer, 20 sl of 0-mercaptoethanol and 10 sl of proteinase K solution, 10 mg/ml) and incubated at 60*C for one hour with continuous shaking. The homogenate 20 obtained was distributed into two Eppendorf vessels (2 ml) and extracted twice by shaking with the same volume of chloroform/isoamyl alcohol (24:1). For phase separation, centrifugation was carried out at 8000g at room temperature for 15 minutes in each case, The DNA was then precipitated at -70*C for 30 minutes using ice-cold isopropanol. The precipitated DNA was sedimented at 4*C and 10,000 g for 30 minutes 25 and resuspended in 180 ul of TE buffer (Sambrook et al., 1989, Cold Spring Harbor Laboratory Press: ISBN 0-87969-309-6). For further purification, the DNA was treated with NaCl (1.2 M final concentration) and precipitated again at -70*C for 30 minutes using twice the volume of absolute ethanol. After a washing step with 70% ethanol, the DNA was dried and subsequently taken up in 50 pl of H20 + RNAse (50 mg/ml final 30 concentration). The DNA was dissolved overnight at 4 0 C and the RNAse digestion was subsequently carried out at 37*C for I h. Storage of the DNA took place at 4*C. -38- Example 3 Isolation of Total RNA and poly-(A)+ RNA from Plants [001151 For the investigation of transcripts, both total RNA and poly-(A)+ RNA were isolated. RNA is isolated from siliques of Arabidopsis plants according to the following 5 procedure: [001161 RNA preparation from Arabidopsis seeds - "hot" extraction: 1001171 1. Buffers, enzymes, and solutions -2M KCI - Proteinase K 10 - Phenol (for RNA) - Chloroform: Isoamylalcohol (Phenol:choloroform 1:1; pH adjusted for RNA) -4 M LiCl, DEPC-treated - DEPC-treated water 15 - 3M NaOAc, pH 5, DEPC-treated - Isopropanol - 70% ethanol (made up with DEPC-treated water) - Resuspension buffer: 0.5% SDS, 10 mM Tris pH 7.5, 1 mM EDTA made up with DEPC-treated water as this solution 20 can not be DEPC-treated - Extraction Buffer: 0.2M Na Borate 30 mM EDTA 30 mM EGTA 25 1% SDS (250pl of 10% SDS-solution for 2.5 ml buffer) 1% Deoxycholate (25 mg for 2.5 ml buffer) 2% PVPP (insoluble - 50 mg for 2.5 ml buffer) 2% PVP 40K (50 mg for 2.5 ml buffer) 10 mM DTT 30 100 mM 0-Mercaptoethanol (fresh, handle under fume hood - use 35 il of 14.3 M solution for 5 ml buffer) [00118] 2. Extraction [001191 The extraction buffer is heated up to 80*C. Tissues are ground in liquid nitrogen-cooled mortar and transferred tissue powder to 1.5m] tubes. Tissue should be 35 kept frozen until buffer is added, therefore, the samples are transferred with a pre-cooled spatula and the tube is kept in liquid nitrogen at all times. Then 350pl preheated extraction buffer is added (here, for 100mg tissue, buffer volume can be as much as 500 pl for bigger samples) to tube. The tube is vortexed, heated to 80*C for -1 minute, and then kept on ice. Samples are vortexed and ground additionally with electric mortar. 40 1001201 3. Digestion -39- [001211 Proteinase K (0.15 mg/100 mg tissue) is added. Then the samples are vortexed and kept at 37*C for one hour. [00122] First Purification [001231 First, 27 jil 2 M KCI is added, and then samples are chilled on ice for 10 5 minutes. The samples are centrifuged at 12,000 rpm for 10 minutes at room temperature, and then the supernatant is transferred to fresh, RNAase-free tubes. One phenol extraction is performed, followed by a chloroform:isoamyl alcohol extraction. One volume isopropanol is added to supernatant, and the mixture is chilled on ice for 10 minutes. RNA is pelleted by centrifugation (7000 rpm for 10 minutes at room 10 temperature). The RNA pellets are dissolved in Iml 4M LiCI by 10 to 15 minutes vortexing. RNA is pelleted by 5 minutes centrifugation. [00124] Second Purification [00125] The pellets are resuspended in 500p] Resuspension buffer. Then, 500Rl phenol is added, and the samples are vortexed. Then, 250 pl chloroform:isoamylalcohol 15 is added, the samples are vortexed and then centrifuged for 5 minutes. The supernatant is transferred to a fresh tube, and the choloform:isoamylalcohol extraction is repeated until the interface is clear. The supernatant is transferred to a fresh tube, and 1/10 vol 3 M NaOAc, pH 5 and 60 0 pl isopropanol are added. The samples are kept at -20C for 20 minutes or longer. RNA is pelleted by 10 minutes centrifugation, and the pellets are 20 washed once with 70% ethanol. All remaining alcohol is removed before resolving the pellets with 15 to 20 gl DEPC-water. The quantity and quality of RNA are determined by measuring the absorbance of a 1:200 dilution at 260 and 280 nm. 4 0Ag RNA/ml = I OD260 [00126] RNA from wild-type and the wril mutant of Arabidopsis is isolated as 25 described (Hosein, 2001, Plant Mol. Biol. Rep. 19, 65a-65e; Ruuska et al., 2002, Plant Cell 14, 1191-1206). The mRNA is prepared from total RNA, using the Amersham Pharmacia Biotech mRNA purification kit, which utilizes oligo(dT)-cellulose columns. Poly-(A)+ RNA is isolated using Dyna BeadsR (Dynal, Oslo, Norway) following the instructions of the manufacturer's protocol. After determination of the concentration of 30 the RNA or of the poly(A)+ RNA, the RNA is precipitated by addition of 1/10 volumes of 3 M sodium acetate pH 4.6 and 2 volumes of ethanol and stored at -70*C. -40- Example 4 cDNA Library Construction 1001271 For cDNA library construction, first strand synthesis was achieved using Murine Leukemia Virus reverse transcriptase (Roche, Mannheim, Germany) and oligo 5 d(T)-primers, second strand synthesis by incubation with DNA polymerase I, Klenow enzyme and RNAseH digestion at 12*C (2 hours), 16*C (1 hour) and 22*C (1 hour). The reaction was stopped by incubation at 65*C (10 minutes) and subsequently transferred to ice. Double stranded DNA molecules were blunted by T4-DNA-polymerase (Roche, Mannheim) at 37*C (30 minutes). Nucleotides were removed by phenol/chloroform 10 extraction and Sephadex G50 spin columns. EcoRI adapters (Pharmacia, Freiburg, Germany) were ligated to the cDNA ends by T4-DNA-ligase (Roche, 12*C, overnight) and phosphorylated by incubation with polynucleotide kinase (Roche, 37*C, 30 minutes). This mixture was subjected to separation on a low melting agarose gel. DNA molecules larger than 300 base pairs were eluted from the gel, phenol extracted, concentrated on 15 Elutip-D-columns (Schleicher and Schuell, Dassel, Germany) and were ligated to vector arms and packed into lambda ZAPH phages or lambda ZAP-Express phages using the Gigapack Gold Kit (Stratagene, Amsterdam, Netherlands) using material and following the instructions of the manufacturer. 20 Example 5 Identification of LMP Genes of Interest 1001281 Arabidopsis wild type and the wril Arabidopsis mutant were used to identify LMP-encoding genes. The wril mutant is characterized by an 80% reduction in seed storage lipids (Focks & Benning, 1998, Plant Physiol. 118:91-101). The WRI1 gene has 25 been cloned and described (Benning&Cernac, 2002, WO 02/072775 A2). [00129] Other LMP candidate genes were identified by various Arabidopsis thaliana developmental or phytohormone mutants (e.g. obtained from EMS treatment or tDNA knock-out mutants) from the Arabidopsis stock center. These mutants and control wild type plants were grown under standard conditions in growth chambers and screened for 30 the accumulation of seed storage compounds. Mutants showing altered levels of seed storage compounds were considered as having a mutation in an LMP candidate gene and were investigated further. The sequences disclosed herein can comprise sequences -41encoding proteins and/or nucleic acids that affect the lipid composition and/or level in a plant. These can be independent of wri 1 or they can also be targets of wri 1 in that they are affected by expression of wril. That effect can be either a decreased oil level or an increased oil level, or an alteration in the oil composition of a plant or part of a plant. 5 Example 6 Cloning offull-length cDNAs of identified LMP genes [00130] Full-length cDNAs were isolated by RACE PCR using the SMART RACE cDNA amplification kit from Clontech allowing both 5'- and 3' rapid amplification of 10 cDNA ends (RACE). The RACE PCR primers were designed based on the proprietary clone sequences. The isolation of full-length cDNAs and the RACE PCR protocol used were based on the manufacturer's conditions. The RACE product fragments were extracted from agarose gels with a QlAquick Gel Extraction Kit (Qiagen) and ligated into the TOPO pCR 2.1 vector (Invitrogen) following manufacturer's instructions. 15 Recombinant vectors were transformed into TOP10 cells (Invitrogen) using standard conditions (Sambrook et al. 1989). Transformed cells were grown overnight at 37"C on LB agar containing 50 pg/mi kanamycin and spread with 40 pl of a 40 mg/ml stock solution of X-gal in dimethylformamide for blue-white selection. Single white colonies were selected and used to inoculate 3 ml of liquid LB containing 50 pg/ml kanamycin 20 and grown overnight at 37*C. Plasmid DNA is extracted using the QlAprep Spin Miniprep Kit (Qiagen) following manufacturer's instructions. Subsequent analyses of clones and restriction mapping was performed according to standard molecular biology techniques (Sambrook et al. 1989). 1001311 Full-length cDNAs were isolated and cloned into binary vectors by using the 25 following procedure: Gene specific primers were designed using the full-length sequences obtained from Arabidopsis cDNA libraries or subsequent RACE amplification products. Full-length sequences and genes were amplified utilizing cDNA libraries as DNA template using touch-down PCR. In some cases, primers were designed to add an "AACA" Kozak-like sequence just upstream of the gene start codon and two bases 30 downstream were, in some cases, changed to GC to facilitate increased gene expression levels (Chandrashekhar et al. 1997, Plant Molecular Biology 35:993-1001). PCR reaction cycles were: 94 *C, 5 minutes; 9 cycles of 94 *C, 1 minute, 65 *C, I minute, 72 *C, 4 minutes and in which the anneal temperature was lowered by 1 "C each cycle; 20 -42cycles of 94 *C, 1 minute, 55 *C, 1 minute, 72 *C, 4 minutes; and the PCR cycle was ended with 72 *C, 10 minutes. Amplified PCR products were gel purified from 1% agarose gels using GenElute -EtBr spin columns (Sigma) and after standard enzymatic digestion, were ligated into the plant binary vector pBPS-GB1 for transformation of 5 Arabidopsis. The binary vector was amplified by overnight growth in E. coli DH5 in LB media and appropriate antibiotic and plasmid was prepared for downstream steps using Qiagen MiniPrep DNA preparation kit. The insert was verified throughout the various cloning steps by determining its size through restriction digest and inserts were sequenced to ensure the expected gene was used in Arabidopsis transformation. 10 1001321 Gene sequences can be used to identify homologous or heterologous genes (orthologs, the same LMP gene from another plant) from cDNA or genomic libraries. This can be done by designing PCR primers to conserved sequences identified by multiple sequence alignments. Orthologs are often identified by designing degenerate primers to full-length or partial sequences of genes of interest. 15 [001331 Gene sequences can be used to identify homologues or orthologs from cDNA or genomic libraries. Homologous genes (e. g. full-length cDNA clones) can be isolated via nucleic acid hybridization using for example cDNA libraries: Depending on the abundance of the gene of interest, 100,000 up to 1,000,000 recombinant bacteriophages are plated and transferred to nylon membranes. After denaturation with alkali, DNA is 20 immobilized on the membrane by, e.g., UV cross linking. Hybridization is carried out at high stringency conditions. Aqueous solution hybridization and washing is performed at an ionic strength of I M NaCl and a temperature of 68C. Hybridization probes are generated by e. g. radioactive (32P) nick transcription labeling (High Prime, Roche, Mannheim, Germany). Signals are detected by autoradiography. 25 [00134] Partially homologous or heterologous genes that are related but not identical can be identified in a procedure analogous to the above-described procedure using low stringency hybridization and washing conditions. For aqueous hybridization, the ionic strength is normally kept at 1 M NaCl while the temperature is progressively lowered from 68 to 42*C. 30 100135] Isolation of gene sequences with homology (or sequence identity/similarity) only in a distinct domain of, for example, 10-20 amino acids, can be carried out by using synthetic radiolabeled oligonucleotide probes. Radiolabeled oligonucleotides are prepared by phosphorylation of the 5-prime end of two complementary oligonucleotides with T4 polynucleotide kinase. The complementary oligonucleotides are annealed and -43ligated to form concatemers. The double stranded concatemers are than radiolabeled by for example nick transcription. Hybridization is normally performed at low stringency conditions using high oligonucleotide concentrations. 5 [001361 Oligonucleotide hybridization solution: o 6 x SSC o 0.01 M sodium phosphate o 1 mM EDTA (pH 8) o 0.5 % SDS 10 o 100 pg/ml denaturated salmon sperm DNA o 0.1 % nonfat dried milk 1001371 During hybridization, temperature is lowered stepwise to 5-10'C below the estimated oligonucleotide Tm or down to room temperature followed by washing steps and autoradiography. Washing is performed with low stringency such as 3 washing steps 15 using 4x SSC. Further details are described by Sambrook et al. (1989, "Molecular Cloning: A Laboratory Manual." Cold Spring Harbor Laboratory Press) or Ausubel et al. (1994, "Current Protocols in Molecular Biology", John Wiley & Sons). [00138] Table 3 20 A table of putative functions of the LMPs (Full length cDNA sequences can be found in the Appendix using the LMP name.) LMP Function ORF position pk309 acyl-(acyl carrier protein) thioesterase 214-1299 pk310 Mitochondrial import inner membrane translocase subunit 26-604 pk3l1 unknown protein 31-1218 pk312 glycosyl transferase, putative 38-1714 pk313 RNA binding like protein 161-1464 pk314 Cdc-45 like protein 294-2081 pk315 F-box family protein-related 1-975 pk316 putative pyruvate kinase, plastid isozyme 44-1756 pk317 pyruvate kinase 135-1922 pk318 Hexokinase 1-1488 pk319 pyruvate kinase 138-1874 Example 7 Identification of Genes of Interest by Screening Expression Libraries with Antibodies 25 1001391 c-DNA clones can be used to produce recombinant protein for example in E. coli (e.g. Qiagen QlAexpress pQE system). Recombinant proteins are then normally -44affinity purified via Ni-NTA affinity chromatography (Qiagen). Recombinant proteins can be used to produce specific antibodies for example by using standard techniques for rabbit immunization. Antibodies are affinity purified using a Ni-NTA column saturated with the recombinant antigen as described by Gu et al. (1994, BioTechniques 17:257 5 262). The antibody can then be used to screen expression cDNA libraries to identify homologous or heterologous genes via an immunological screening (Sambrook et al. 1989, Molecular Cloning: A Laboratory Manual," Cold Spring Harbor Laboratory Press or Ausubel et al., 1994, "Current Protocols in Molecular Biology," John Wiley & Sons). 10 Example 8 Northern-Hybridization 1001401 For RNA hybridization, 20 sLg of total RNA or 1 sg of poly-(A)+ RNA is separated by gel electrophoresis in 1.25% agarose gels using formaldehyde as described in Amasino (1986, Anal. Biochem. 152:304), transferred by capillary attraction using 10 15 x SSC to positively charged nylon membranes (Hybond N+, Amersham, Braunschweig), immobilized by UV light and pre-hybridized for 3 hours at 68 0 C using hybridization buffer (10% dextran sulfate w/v, I M NaCl, 1% SDS, 100 sg/ml of herring sperm DNA). The labeling of the DNA probe with the Highprime DNA labeling kit (Roche, Mannheim, Germany) is carried out during the pre-hybridization using alpha-32P dCTP 20 (Amersham, Braunschweig, Germany). Hybridization is carried out after addition of the labeled DNA probe in the same buffer at 68*C ovemight. The washing steps are carried out twice for 15 minutes using 2 x SSC and twice for 30 minutes using I x SSC, 1% SDS at 68*C. The exposure of the sealed filters is carried out at -70*C for a period of 1 day to 14 days. 25 Example 9 DNA Sequencing and Computational Functional Analysis [001411 cDNA libraries can be used for DNA sequencing according to standard methods, in particular by the chain termination method using the ABI PRISM Big Dye Terminator Cycle Sequencing Ready Reaction Kit (Perkin-Elmer, Weiterstadt, 30 Germany). Random sequencing can be carried out subsequent to preparative plasmid recovery from cDNA libraries via in vivo mass excision, retransformation, and subsequent plating of DH1OB on agar plates (material and protocol details from -45- Stratagene, Amsterdam, Netherlands). Plasmid DNA can be prepared from overnight grown E. coli cultures grown in Luria-Broth medium containing ampicillin (See Sambrook et al. (1989, Cold Spring Harbor Laboratory Press: ISBN 0-87969-309-6) on a Qiagene DNA preparation robot (Qiagen, Hilden) according to the manufacturer's 5 protocols). Sequences can be processed and annotated using the software package EST MAX commercially provided by Bio-Max (Munich, Germany). The program incorporates bioinformatics methods important for functional and structural characterization of protein sequences. For reference, see http://pedant.mlps.biochem.mpg.de. 10 1001421 The most important algorithms incorporated in EST-MAX are: FASTA: Very sensitive protein sequence database searches with estimates of statistical significance (Pearson, 1990, Rapid and sensitive sequence comparison with FASTP and FASTA. Methods Enzymol, 183:63-98); BLAST: Very sensitive protein sequence database searches with estimates of statistical significance (Altschul et al., Basic local alignment 15 search tool. J. Mol. Biol. 215:403-410); PREDATOR: High-accuracy secondary structure prediction from single and multiple sequences (Frishman & Argos, 1997, 75% accuracy in protein secondary structure prediction. Proteins 27:329-335); CLUSTALW: Multiple sequence alignment (Thompson et al., 1994, CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific 20 gap penalties and weight matrix choice, Nucleic Acids Res. 22:4673-4680); TMAP: Transmembrane region prediction from multiply aligned sequences (Persson & Argos, 1994, Prediction of transmembrane segments in proteins utilizing multiple sequence alignments, J. Mol. Biol. 237:182-192); ALOM2:Transmembrane region prediction from single sequences (Klein et al., 1984, Prediction of protein function from sequence 25 properties: A discriminant analysis of a database. Biochim. Biophys. Acta 787:221-226. Version 2 by Dr. K. Nakai); PROSEARCH: Detection of PROSITE protein sequence patterns. Kolakowski et al., 1992, ProSearch: fast searching of protein sequences with regular expression patterns related to protein structure and function. Biotechniques 13:919-921); BLIMPS: Similarity searches against a database of ungapped blocks 30 (Wallace & Henikoff, 1992, PATMAT: A searching and extraction program for sequence, pattern and block queries and databases, CABIOS 8:249-254. Written by Bill Alford). -46- Example 10 Plasmids for Plant Transformation [001431 For plant transformation binary vectors such as pBinAR can be used (H6fgen & Willmitzer 1990, Plant Sci. 66:221-230). Construction of the binary vectors can be 5 performed by ligation of the cDNA in sense or antisense orientation into the T-DNA. 5' to the cDNA a plant promoter activates transcription of the cDNA. A polyadenylation sequence is located 3' to the cDNA. Tissue-specific expression can be achieved by using a tissue specific promoter. For example, seed-specific expression can be achieved by cloning the napin or LeB4 or USP promoter 5' to the cDNA. Also, any other seed 10 specific promoter element can be used. For constitutive expression within the whole plant, the CaMV 35S promoter can be used. The expressed protein can be targeted to a cellular compartment using a signal peptide, for example for plastids, mitochondria, or endoplasmic reticulum (Kermode, 1996, Crit. Rev. Plant Sci. 15:285-423). The signal peptide is cloned 5' in frame to the cDNA to achieve subcellular localization of the 15 fusion protein. [00144] Further examples for plant binary vectors are the pBPS-GBI, pSUN2-GW, or pBPS-GB047 vectors into which the LMP gene candidates are cloned. These binary vectors contain an antibiotic resistance gene driven under the control of the AtAct2-I promoter and a seed-specific promoter or a constitutive promoter in front of the candidate 20 gene with the NOSpA terminator or the OCS terminator. Partial or full-length LMP cDNAs are cloned into the multiple cloning site of the plant binary vector in sense or antisense orientation behind the USP or other seed-specific, tissue-specific, or constitutive promoters. The recombinant vector containing the gene of interest is transformed into ToplO cells (Invitrogen) using standard conditions. Transformed cells 25 are selected for on LB agar containing 50 pg/ml kanamycin grown overnight at 37"C. Plasmid DNA is extracted using the QlAprep Spin Miniprep Kit (Qiagen) following manufacturer's instructions. Analysis of subsequent clones and restriction mapping is performed according to standard molecular biology techniques (Sambrook et al. 1989, Molecular Cloning, A Laboratory Manual. 2 nd Edition. Cold Spring Harbor Laboratory 30 Press. Cold Spring Harbor, NY). -47- Example 11 Agrobacterium Mediated Plant Transformation [00145] Agrobacterium mediated plant transformation with the LMP nucleic acids described herein can be performed using standard transformation and regeneration 5 techniques (Gelvin & Schilperoort, Plant Molecular Biology Manual, 2nd ed. Kluwer Academic Pub]., Dordrecht 1995 in Sect., Ringbuc Zentrale Signatur:BTI -P; Glick, Bernard R. and Thompson, John E. Methods in Plant Molecular Biology and Biotechnology, S. 360, CRC Press, Boca Raton 1993). For example, Agrobacterium mediated transformation can be performed using the GV3 (pMP90) (Koncz & Schell, 10 1986, Mol. Gen. Genet. 204:383-396) or LBA4404 (Clontech) Agrobacterium tumefaciens strain. [001461 Arabidopsis thaliana can be grown and transformed according to standard conditions (Bechtold 1993, Acad. Sci. Paris. 316:1194-1199; Bent et al. 1994, Science 265:1856-1860). Additionally, rapeseed can be transformed with the LMP nucleic acids 15 of the present invention via cotyledon or hypocotyl transformation (Moloney et al. 1989, Plant Cell Report 8:238-242; De Block et al. 1989, Plant Physiol. 91:694-701). Use of antibiotics for Agrobacterium and plant selection depends on the binary vector and the Agrobacterium strain used for transformation. Rapeseed selection is normally performed using kanamycin as selectable plant marker. Additionally, Agrobacterium mediated gene 20 transfer to flax can be performed using, for example, a technique described by Mlynarova et al. (1994, Plant Cell Report 13:282-285). [001471 Transformation of soybean can be performed using for example a technique described in EP 0424 047, U.S. Patent No. 5,322,783 (Pioneer Hi-Bred International) or in EP 0397 687, U.S. Patent No. 5,376,543 or U.S. Patent No. 5,169,770 (University 25 Toledo). Soybean seeds are surface sterilized with 70% ethanol for 4 minutes at room temperature with continuous shaking, followed by 20% (v/v) CLOROX supplemented with 0.05% (v/v) TWEEN for 20 minutes with continuous shaking. Then the seeds are rinsed 4 times with distilled water and placed on moistened sterile filter paper in a Petri dish at room temperature for 6 to 39 hours. The seed coats are peeled off, and cotyledons 30 are detached from the embryo axis. The embryo axis is examined to make sure that the meristematic region is not damaged. The excised embryo axes are collected in a half open sterile Petri dish and air-dried to a moisture content less than 20% (fresh weight) in a sealed Petri dish until further use. -48- [001481 The method of plant transformation is also applicable to Brassica and other crops. In particular, seeds of canola are surface sterilized with 70% ethanol for 4 minutes at room temperature with continuous shaking, followed by 20% (v/v) CLOROX supplemented with 0.05 % (v/v) TWEEN for 20 minutes, at room temperature with 5 continuous shaking. Then, the seeds are rinsed 4 times with distilled water and placed on moistened sterile filter paper in a Petri dish at room temperature for 18 hours. The seed coats are removed and the seeds are air dried overnight in a half-open sterile Petri dish. During this period, the seeds lose approximately 85% of their water content. The seeds are then stored at room temperature in a sealed Petri dish until further use. 10 1001491 Agrobacterium tumefaciens culture is prepared from a single colony in LB solid medium plus appropriate antibiotics (e.g. 100 mg/l streptomycin, 50 mg/l kanamycin) followed by growth of the single colony in liquid LB medium to an optical density at 600 nm of 0.8. Then, the bacteria culture is pelleted at 7000 rpm for 7 minutes at room temperature, and re-suspended in MS (Murashige & Skoog 1962, Physiol. Plant. 15 15:473-497) medium supplemented with 100 mM acetosyringone. Bacteria cultures are incubated in this pre-induction medium for 2 hours at room temperature before use. The axis of soybean zygotic seed embryos at approximately 44% moisture content are imbibed for 2 hours at room temperature with the pre-induced Agrobacterium suspension culture. (The imbibition of dry embryos with a culture of Agrobacterium is also 20 applicable to maize embryo axes). 1001501 The embryos are removed from the imbibition culture and are transferred to Petri dishes containing solid MS medium supplemented with 2% sucrose and incubated for 2 days, in the dark at room temperature. Alternatively, the embryos are placed on top of moistened (liquid MS medium) sterile filter paper in a Petri dish and incubated under 25 the same conditions described above. After this period, the embryos are transferred to either solid or liquid MS medium supplemented with 500 mg/l carbenicillin or 300 mg/l cefotaxime to kill the agrobacteria. The liquid medium is used to moisten the sterile filter paper. The embryos are incubated during 4 weeks at 25*C, under 440 jymol m- 2 s- 1 and 12 hours photoperiod. Once the seedlings have produced roots, they are transferred to 30 sterile metromix soil. The medium of the in vitro plants is washed off before transferring the plants to soil. The plants are kept under a plastic cover for 1 week to favor the acclimatization process. Then the plants are transferred to a growth room where they are -49incubated at 25*C, under 440 ymol m 2 s- 1 light intensity and 12 h photoperiod for about 80 days. 1001511 Samples of the primary transgenic plants (TO) are analyzed by PCR to confirm the presence of T-DNA. These results are confirmed by Southern hybridization wherein 5 DNA is electrophoresed on a 1% agarose gel and transferred to a positively charged nylon membrane (Roche Diagnostics). The PCR DIG Probe Synthesis Kit (Roche Diagnostics) is used to prepare a digoxigenin-labeled probe by PCR as recommended by the manufacturer. Example 12 10 In vivo Mutagenesis [001521 In vivo mutagenesis of microorganisms can be performed by incorporation and passage of the plasmid (or other vector) DNA through E. coli or other microorganisms (e.g. Bacillus spp. or yeasts such as Saccharomyces cerevisiae),that are impaired in their capabilities to maintain the integrity of their genetic information. 15 Typical mutator strains have mutations in the genes for the DNA repair system (e.g., mutHLS, mutD, mutT, etc.; for reference, see Rupp, 1996, DNA repair mechanisms, in: Escherichia coli and Salmonella, p. 2277-2294, ASM: Washington.) Such strains are well known to those skilled in the art. The use of such strains is illustrated, for example, in Greener and Callahan 1994, Strategies 7:32-34. Transfer of mutated DNA molecules 20 into plants is preferably done after selection and testing in microorganisms. Transgenic plants are generated according to various examples within this document. Example 13 Assessment of the mRNA Expression and Activity of a Recombinant Gene Product in the Transformed Organism 25 [00153] The activity of a recombinant gene product in the transformed host organism can be measured on the transcriptional and/or on the translational level. A useful method to ascertain the level of transcription of the gene (an indicator of the amount of mRNA available for translation to the gene product) is to perform a Northcrn blot (for reference see, for example, Ausubel et al., 1988, Current Protocols in Molecular Biology, Wiley: 30 New York), in which a primer designed to bind to the gene of interest is labeled with a detectable tag (usually radioactive or chemiluminescent), such that when the total RNA of a culture of the organism is extracted, run on gel, transferred to a stable matrix and incubated with this probe, the binding and quantity of binding of the probe indicates the -50presence and also the quantity of mRNA for this gene. This information at least partially demonstrates the degree of transcription of the transformed gene. Total cellular RNA can be prepared from plant cells, tissues, or organs by several methods, all well-known in the art, such as that described in Bormann et al. (1992, Mol. Microbiol. 6:317-326). 5 [001541 To assess the presence or relative quantity of protein translated from this mRNA, standard techniques, such as a Western blot, may be employed (see, for example, Ausubel et al. 1988, Current Protocols in Molecular Biology, Wiley: New York). In this process, total cellular proteins are extracted, separated by gel electrophoresis, transferred to a matrix such as nitrocellulose, and incubated with a probe, such as an antibody, which 10 specifically binds to the desired protein. This probe is generally tagged with a chemiluminescent or colorimetric label, which may be readily detected. The presence and quantity of label observed indicates the presence and quantity of the desired mutant protein present in the cell. [001551 The activity of LMPs that bind to DNA can be measured by several well 15 established methods, such as DNA band-shift assays (also called gel retardation assays). The effect of such LMP on the expression of other molecules can be measured using reporter gene assays (such as that described in Kolmar et al., 1995, EMBO J. 14:3895 3904 and references cited therein). Reporter gene test systems are well known and established for applications in both prokaryotic and eukaryotic cells, using enzymes such 20 as beta-galactosidase, green fluorescent protein, and several others. [00156] The determination of activity of lipid metabolism membrane-transport proteins can be performed according to techniques such as those described in Gennis R.B. (1989 Pores, Channels and Transporters, in Biomembranes, Molecular Structure and Function, Springer: Heidelberg, pp. 85-137, 199-234 and 270-322). 25 Example 14 In vitro Analysis of the Function ofArabidopsis thaliana Genes in Transgenic Plants [001571 The determination of activities and kinetic parameters of enzymes is well established in the art. Experiments to determine the activity of any given altered enzyme 30 must be tailored to the specific activity of the wild-type enzyme, which is well within the ability of one skilled in the art. Overviews about enzymes in general, as well as specific details concerning structure, kinetics, principles, methods, applications and examples for the determination of many enzyme activities may be found, for example, in the following -51references: Dixon & Webb, 1979, Enzymes. Longmans: London; Fersht, (1985) Enzyme Structure and Mechanism. Freeman: New York; Walsh (1979) Enzymatic Reaction Mechanisms. Freeman: San Francisco; Price, N.C., Stevens, L. (1982) Fundamentals of Enzymology. Oxford Univ. Press: Oxford; Boyer, P.D., ed. (1983) The Enzymes, 3rd ed. 5 Academic Press: New York; Bisswanger, H., (1994) Enzymkinetik, 2nd ed. VCH: Weinheim (ISBN 3527300325); Bergmeyer, H.U., Bergmeyer, J., Gral, M., eds. (1983 1986) Methods of Enzymatic Analysis, 3rd ed., vol. I-XII, Verlag Chemie: Weinheim; and Ullmann's Encyclopedia of Industrial Chemistry (1987) vol. A9, Enzymes. VCH: Weinheim, p. 352-363. 10 Example 15 Analysis ofthe Impact of Recombinant Proteins on the Production of a Desired Seed Storage Compound 100158) The effect of the genetic modification in plants on a desired seed storage compound (such as a sugar, lipid, or fatty acid) can be assessed by growing the modified 15 plant under suitable conditions and analyzing the seeds or any other plant organ for increased production of the desired product (i.e., a lipid or a fatty acid). Such analysis techniques are well known to one skilled in the art, and include spectroscopy, thin layer chromatography, staining methods of various kinds, enzymatic and microbiological methods, and analytical chromatography such as high performance liquid 20 chromatography (see, for example, Ullman 1985, Encyclopedia of Industrial Chemistry, vol. A2, pp. 89-90 and 443-613, VCH: Weinheim; Fallon et al., 1987, Applications of HPLC in Biochemistry in: Laboratory Techniques in Biochemistry and Molecular Biology, vol. 17; Rehm et al., 1993, Product recovery and purification, Biotechnology, vol. 3, Chapter III, pp. 469-714, VCH: Weinheim; Belter, P.A. et al., 1988 25 Bioseparations: downstream processing for biotechnology, John Wiley & Sons; Kennedy & Cabral, 1992, Recovery processes for biological materials, John Wiley and Sons; Shaeiwitz & Henry, 1988, Biochemical separations in: Ulmann's Encyclopedia of Industrial Chemistry, Separation and purification techniques in biotechnology, vol. B3, Chapter 11, pp. 1-27, VCH: Weinheim; and Dechow F.J. 1989). 30 [00159] Besides the above-mentioned methods, plant lipids are extracted from plant material as described by Cahoon et al. (1999, Proc. Nat]. Acad. Sci. USA 96, 22:12935 12940) and Browse et al. (1986, Anal. Biochemistry 442:141-145). Qualitative and quantitative lipid or fatty acid analysis is described in Christie, William W., Advances in -52- Lipid Methodology. Ayr/Scotland:Oily Press. - (Oily Press Lipid Library; Christie, William W., Gas Chromatography and Lipids. A Practical Guide - Ayr, Scotland:Oily Press, 1989 Repr. 1992. - IX,307 S. - (Oily Press Lipid Library; and "Progress in Lipid Research, Oxford :Pergamon Press, 1 (1952) - 16 (1977) Progress in the Chemistry of 5 Fats and Other Lipids CODEN. [001601 Unequivocal proof of the presence of fatty acid products can be obtained by the analysis of transgenic plants following standard analytical procedures: GC, GC-MS or TLC as described by Christie and references therein (1997 in: Advances on Lipid Methodology 4th ed.: Christie, Oily Press, Dundee, pp. 119-169; 1998). Detailed 10 methods are described for leaves by Lemieux et al. (1990, Theor. Apple. Genet. 80:234 240) and for seeds by Focks & Benning (1998, Plant Physiol. 118:91-10 1). [001611 Positional analysis of the fatty acid composition at the sn-1, sn-2 or sn-3 positions of the glycerol backbone is determined by lipase digestion (See, e.g., Siebertz & Heinz 1977, Z. Naturforsch. 32c:193-205, and Christie 1987, Lipid Analysis 2 nd Edition, 15 Pergamon Press, Exeter, ISBN 0-08-023791-6). [001621 Total seed oil levels can be measured by any appropriate method. Quantitation of seed oil contents is often performed with conventional methods, such as near infrared analysis (NIR) or nuclear magnetic resonance imaging (NMR). NTR spectroscopy has become a standard method for screening seed samples whenever the samples of interest 20 have been amenable to this technique. Samples studied include canola, soybean, maize, wheat, rice, and others. NIR analysis of single seeds can be used (See, e.g., Velasco et al., 'Estimation of seed weight, oil content and fatty acid composition in intact single seeds of rapeseed (Brassica napus L.) by near-infrared reflectance spectroscopy, 'Euphytica, Vol. 106, 1999, pp. 79-85). NMR has also been used to analyze oil content 25 in seeds (See, e.g., Robertson & Morrison, "Analysis of oil content of sunflower seed by wide-line NMR, "Journal of the American Oil Chemists Society, 1979, Vol. 56, 1979, pp. 961-964, which is herein incorporated by reference in its entirety). [00163] A typical way to gather information regarding the influence of increased or decreased protein activities on lipid and sugar biosynthetic pathways is for example via 30 analyzing the carbon fluxes by labeling studies with leaves or seeds using "C-acetate or 14 C-pyruvate (See, e.g., Focks & Benning, 1998, Plant Physiol. 118:91-101; Eccleston & Ohlrogge, 1998, Plant Cell 10:613-621). The distribution of 1C into lipids and aqueous soluble components can be determined by liquid scintillation counting after the respective -53separation (for example on TLC plates) including standards like 1C-sucrose and "C malate (Eccleston & Ohlrogge, 1998, Plant Cell 10:613-621). [001641 Material to be analyzed can be disintegrated via sonification, glass milling, liquid nitrogen and grinding or via other applicable methods. The material has to be 5 centrifuged after disintegration. The sediment is resuspended in distilled water, heated for 10 minutes at 100*C, cooled on ice, and centrifuged again, followed by extraction in 0.5 M sulfuric acid in methanol containing 2% dimethoxypropane for 1 hour at 90*C leading to hydrolyzed oil and lipid compounds, resulting in transmethylated lipids. These fatty acid methyl esters are extracted in petrolether and finally subjected to GC analysis 10 using a capillary column (Chrompack, WCOT Fused Silica, CP-Wax-52 CB, 25 m, 0.32 mm) at a temperature gradient between 170*C and 240'C for 20 minutes, and then 5 minutes at 240*C. The identity of resulting fatty acid methylesters is defined by the use of standards available form commercial sources (i.e., Sigma). In case of fatty acids where standards are not available, molecule identity is shown via derivatization and 15 subsequent GC-MS analysis. For example, the localization of triple bond fatty acids is shown via GC-MS after derivatization via 4,4-Dimethoxy-oxazolin-Derivaten (Christie, Oily Press, Dundee, 1998). [00165] A common standard method for analyzing sugars, especially starch, is published by Stitt et al. (1989, Methods Enzymol. 174:518-552; for other methods, see 20 also Hartel et al., 1998, Plant Physiol. Biochem. 36:407-417 and Focks & Benning, 1998, Plant Physiol. 118:91-10 1). 100166] For the extraction of soluble sugars and starch, 50 seeds are homogenized in 500 pl of 80% (v/v) ethanol in a 1.5-ml polypropylene test tube and incubated at 70*C for 90 minutes. Following centrifugation at 16,000 g for 5 minutes, the supernatant is 25 transferred to a new test tube. The pellet is extracted twice with 500 Al of 80% ethanol. The solvent of the combined supernatants is evaporated at room temperature under a vacuum. The residue is dissolved in 50 pl of water, representing the soluble carbohydrate fraction. The pellet left from the ethanol extraction, which contains the insoluble carbohydrates including starch, is homogenized in 200 pl of 0.2 N KOH, and the 30 suspension is incubated at 95 0 C for I hour to dissolve tie starch. Following the addition of 35 pil of 1 N acetic acid and centrifugation for 5 minutes at 16,000 g, the supernatant is used for starch quantification. -54- 1001671 To quantify soluble sugars, 10 pl of the sugar extract is added to 990 Al of reaction buffer containing 100 mM imidazole, pH 6.9, 5 mM MgCI2, 2 mM NADP, I mM ATP, and 2 units 2 ml- of Glucose-6-P-dehydrogenase. For enzymatic determination of glucose, fructose and sucrose, 4.5 units of hexokinase, 1 unit of 5 phosphoglucoisomerase, and 2 Al of a saturated fructosidase solution are added in succession. The production of NADPH is photometrically monitored at a wavelength of 340 rn. Similarly, starch is assayed in 30 ul of the insoluble carbohydrate fraction with a kit from Boehringer Mannheim. 1001681 An example for analyzing the protein content in leaves and seeds can be found 10 by Bradford (1976, Anal. Biochem. 72:248-254). For quantification of total seed protein, 15-20 seeds are homogenized in 250 Al of acetone in a 1.5-ml polypropylene test tube. Following centrifugation at 16,000 g, the supernatant is discarded, and the vacuum-dried pellet is resuspended in 250 yI1 of extraction buffer containing 50 mM Tris-HCl, pH 8.0, 250 mM NaCl, 1 mM EDTA, and 1% (w/v) SDS. Following incubation for 2 hours at 15 25*C, the homogenate is centrifuged at 16,000 g for 5 minutes, and 200 ml of the supernatant will be used for protein measurements. In the assay, y-globulin is used for calibration. For protein measurements, Lowry DC protein assay (Bio-Rad) or Bradford assay (Bio-Rad) is used. [001691 Enzymatic assays of hexokinase and fructokinase are performed 20 spectrophotometrically according to Renz et al. (1993, Planta 190:156-165), of phosphogluco-isomerase, ATP-dependent 6-phosphofructokinase, pyrophosphate dependent 6-phospho-fructokinase, Fructose-1,6-bisphosphate aldolase, triose phosphate isomerase, glyceral-3-P dehydrogenase, phosphoglycerate kinase, phosphoglycerate mutase, enolase, and pyruvate kinase are performed according to Burrell et al. (1994, 25 Planta 194:95-101) and of UDP-Glucose-pyrophosphorylase according to Zrenner et al. (1995, Plant J. 7:97-107). 1001701 Intermediates of the carbohydrate metabolism, like Glucose-l-phosphate, Glucose-6-phosphate, Fructose-6-phosphate, Phosphoenolpyruvate, Pyruvate, and ATP are measured as described in Hartel et al. (1998, Plant Physiol. Biochem. 36:407-417) 30 and metabolites are measured as described in Jelitto et al. (1992, Planta 188:238-244). 100171] In addition to the measurement of the final seed storage compound (i.e., lipid, starch or storage protein) it is also possible to analyze other components of the metabolic pathways utilized for the production of a desired seed storage compound, such as -55intermediates and side-products, to determine the overall efficiency of production of the compound (Fiehn et al., 2000, Nature Biotech. 18:1447-1161). For example, yeast expression vectors comprising the nucleic acids disclosed herein, or fragments thereof, can be constructed and transformed into Saccharomyces cerevisiae using standard 5 protocols. The resulting transgenic cells can then be assayed for alterations in sugar, oil, lipid, or fatty acid contents. [001721 Similarly, plant expression vectors comprising the nucleic acids disclosed herein, or fragments thereof, can be constructed and transformed into an appropriate plant cell such as Arabidopsis, soybean, rapeseed, rice, maize, wheat, Medicago truncalula, 10 etc., using standard protocols. The resulting transgenic cells and/or plants derived from the cells can then be assayed for alterations in sugar, oil, lipid, or fatty acid contents. [001731 Additionally, the sequences disclosed herein, or fragments thereof, can be used to generate knockout mutations in the genomes of various organisms, such as bacteria, mammalian cells, yeast cells, and plant cells (Girke at al., 1998, Plant J. 15:39 15 48). The resultant knockout cells can then be evaluated for their composition and content in seed storage compounds, and the effect on the phenotype and/or genotype of the mutation. Other methods of gene inactivation include those described in US 6,004,804 and Puttaraju et al. (1999, Nature Biotech. 17:246-252). 20 Example 16 Purification of the Desired Product from Transformed Organisms 1001741 An LMP can be recovered from plant material by various methods well known in the art. Organs of plants can be separated mechanically from other tissue or organs prior to isolation of the seed storage compound from the plant organ. Following 25 homogenization of the tissue, cellular debris is removed by centrifugation and the supernatant fraction containing the soluble proteins is retained for further purification of the desired compound. If the product is secreted from cells grown in culture, then the cells are removed from the culture by low-speed centrifugation, and the supemate fraction is retained for further purification. 30 [001751 The supernatant fraction from either purification method is subjected to chromatography with a suitable resin, in which the desired molecule is either retained on a chromatography resin while many of the impurities in the sample are not, or where the impurities are retained by the resin, while the sample is not. Such chromatography steps -56may be repeated as necessary, using the same or different chromatography resins. One skilled in the art would be well-versed in the selection of appropriate chromatography resins and in their most efficacious application for a particular molecule to be purified. The purified product may be concentrated by filtration or ultrafiltration, and stored at a 5 temperature at which the stability of the product is maximized. 1001761 There is a wide array of purification methods known to the art and the preceding method of purification is not meant to be limiting. Such purification techniques are described, for example, in Bailey & Ollis, 1986, Biochemical Engineering Fundamentals, McGraw-Hill:New York). 10 [001771 The identity and purity of the isolated compounds may be assessed by techniques standard in the art. These include high-performance liquid chromatography (HPLC), spectroscopic methods, staining methods, thin layer chromatography, analytical chromatography such as high performance liquid chromatography, NIRS, enzymatic assay, or microbiologically. Such analysis methods are reviewed in: Patek et al. (1994, 15 Appl. Environ. Microbiol. 60:133-140), Malakhova et al. (1996, Biotekhnologiya 11:27 32), and Schmidt et al. (1998, Bioprocess Engineer 19:67-70), Ulmann's Encyclopedia of Industrial Chemistry (1996, Vol. A27, VCH: Weinheim, p. 89-90, p. 521-540, p. 540 547, p. 559-566, 575-581 and p. 581-587), and Michal G. (1999, Biochemical Pathways: An Atlas of Biochemistry and Molecular Biology, John Wiley and Sons; Fallon, A. et al. 20 1987, Applications of HPLC in Biochemistry in: Laboratory Techniques in Biochemistry and Molecular Biology, vol. 17). Example 17 Screening for increased stress tolerance and plant growth 25 [001781 The transgenic plants are screened for their improved stress tolerance demon strating that transgene expression confers stress tolerance. The transgenic plants are further screened for their growth rate demonstrating that transgene expression confers increased growth rates and/or increased seed yield. 1001791 Increased seed size might be reflected in an increased seed weight of gene 30 overexpressors. Increased seed size leads to greater yield in many economically important crop plants. Therefore, increased seed size is one goal of genetically engineering and selection using LMPs as described in this application. -57- [001801 For in vitro root analysis square plates measuring 12 cm x 12 cm can be used. For each plate, 52 ml of MS media (0.5X MS salts, 0.5% sucrose, 0.5 g/L MES buffer, 1% Phytagar) without selection will be used. Plates will be allowed to dry in the sterile hood for one hour to reduce future condensation. Seed aliquots will be sterilized in glass 5 vials with ethanol for 5 minutes, ethanol will be removed, and the seeds are allowed to dry in the sterile hood for one hour. [001811 Seeds will be spotted in the plates using the Vacuseed Device (Lehle). After the seeds are spotted on the plates, the plates will be wrapped with Ventwrap and placed vertically in racks in the dark at 4*C for four days to stratify the seeds. The plates are 10 transferred to a C5 Percival Growth Chamber and placed vertically. The growth chamber conditions will be 23*C day/21*C night and 16 hour day/8 hour night. For data collection, a high-resolution flatbed scanner is used. Analysis of the roots is done using the WinRhizo software package. [001821 A comparison of the root length obtained with Arabidopsis wild type and the 15 wril mutant indicated a significant reduction in root length in wril mutants. This reduction in root length was found to be associated with a delayed germination and a reduced number of leaves at a defined time point of development as compared with the wild type. Overexpression of genes involved in the WRI1 regulatory network in wild type background may improve seed germination, increase root length, and increase speed 20 of leaf development and number of leaves. The latter may improve photosynthetic performance of plants resulting in increase yield of biomass and in increased amounts and/or size of seeds associated with increased amounts of seed storage compounds like oil, protein, and sugars. [001831 For soil root analysis, seeds may be imbibed at 4*C for 2 days in water and 25 planted directly in soil with no selection. Deepots (Hummert D40) will be used with a saturated peat pellet (Jiffy 727) at the base and filled with water saturated Metromix. After planting, pots will be covered with plastic wrap to prevent drying. Plants may be grown using only water present at media preparation, as the water in the soil in these large pots is sufficient for 3 weeks of growth, and encourages rapid root growth. The 30 plastic wrapping of the pots will be removed after 12 days and morphological data documented, At day 17, the aerial parts of the plant will be harvested, dried (65*C for 2 days), and dry weight measured. To examine the roots the peat pellet will be pushed towards the top of the pot to remove the soil and roots as a unit. The soil will then be -58separated from the roots in a tray and the maximum root length will be measured. Root length of all plants for all transgenic lines will be averaged and compared against the average of the wild type plants. [001841 Those skilled in the art will recognize, or will be able to ascertain using no 5 more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Such equivalents are intended to be encompasses by the claims to the invention disclosed and claimed herein. 10 APPENDIX Nucleic acid sequence of pk309 (SEQ ID NO:1) TGACAATTGTCCATCATCAATGGCATrAAATGGCAAAACCGTAATTTCGA ACTCCACCAAGGGGCAAACTTAAAAGTCGATGTCTTTCTTCTTCACCTCGG ACCCATCGGAGAGAAGATACTACTAGAAGAGATTCATTCACAGTGTTGAA 15 ATTAAAAAACCGAAACTTTCTCGTTFCTTCTTCTTTCTCCAATTTTCAAA ATTCGAAAAGATGTTGAAGCTrTCGTGTAATGTGACTGATTCTAAGTTACA GAGAAGCTrACTCTTCTTCTCCCATTCATATCGATCTGATCCGGTGAATIT CATCCGTCGGAGAATTGTCTCTTGTTCTCAGACGAAGAAGACAGGTTTGGT TCCTTTGCGTGCTGTrGTATCTGCTGATCAAGGAAGTGTGGTTCAAGGTTT 20 GGCTACTCTCGCGGATCAGCTCCGATTAGGTAGTTrGACTGAAGATGGTTT ATCTTATAAAGAGAAGTTTGTTGTTAGATCTTACGAAGTGGGTAGTAACA AAACCGCTACTGTTGAAACCATTGCTAATCTTITACAGGAGGTGGGATGT AATCATGCACAAAGTGTFGGTTTCGACTGATGGGTTTGCAACAACAACT ACTATGAGGAAGTTGCATCTCATTTGGGTTACTGCGAGAATGCATATCGA 25 GATCTATAAGTACCCTGCTTGGGGTGATGTGGTTGAGATAGAGACTTGGT GTCAGAGTGAAGGAAGGATTGGGACAAGGCGTGATTGGATTCTTAAGGAT TCTGTCACTGGTGAAGTCACTGGCCGTGCTACAAGCAAGTGGGTGATGAT GAACCAAGACACGAGACGGCTTCAGAAAGTTTCTGATGATGTTCGGGACG AGTACTTGGTCTTCTGTCCTCAAGAACCGAGGTrAGCATTTCCGGAAGAG 30 AATAACAGAAGCTFGAAGAAAATCCCGAAACTCGAAGATCCGGCTCAGTA TTCAATGATTGGGCTTAAGCCTAGACGAGCTGATCTCGACATGAACCAGC ATGTCAATAATGTCACCTATATTGGATGGGTTCTCGAGAGCATACCACAA GAAATTGTAGACACGCACGAGCTTCAGGTCATAACTCTGGATTATAGAAG AGAATGTCAACAAGACGATGTGGTGGATTCACTCACCACCACCACCTCTG 35 AAATTGGTGGAACCAATGGCTCTGCCACGTCTGGCACACAGGGCCACAAC GATAGCCAGTTCTTGCACCTCCTGAGGTTGTCTGGAGATGGTCAGGAGAT CAACCGCGGGACAACTCTGTGGAGAAAGAAGCCTTCAAGTTAAGAAATA GACAATGTCTTTAGCCATTTTGTTCTCAAGTTTCCATCATCTCAATGAAGA TFCGCTTCACGAGTCTGAGCAGGTCTCCATTTITTCTCTTTCAAGTTTGG 40 GTrAGACTAGAGGGAACTGGATrGTrGGAGTATTAATCTTTGTTGAATTTC ATTATGTITGTTCATG1 TTTGTACAAATTTTGGGGATTTAGCCAAACCCAT ATCGTCTTrGGTCTTGTCTTGTGCCTGCGTGTATTTTTAATCTATCCGTTCA AAACACAGATTGTTCATTAGATGTTATATAAACAGAGTTAAAGACCTG A 45 -59- Nucleic acid sequence of pk310 (SEQ ID NO:2) GAAATTGGAGTTCTCTCGAAGTTCCGTGGCGTCAAAAATGGCGTTGGGTG ATCGGAAATCCCCAGAACAAACAAATCAGGCGTTATCTCCTCCGACGCCT ATTGTGCAGGAAAATGGAACTCCGACGAAGCGTGTGTTGATCACTTCCCT 5 TTrAGCAGGAGTAATTGGTGGAGGAGCTGGTTTAGTGTCTAAACACCGGA TAGCTCATCCCAATATTCCTACTGTTTACGCTGCTAATTTTGCTATTGTCGC CGGTTGCTATTGCGGAGCTCGTGAATCTGTGAGAATAACTCGAAGATCAG AACACGATGATTTAATGAACTCAGCTATTGGAGGACTIT1AGTGGTGCTT TGCTTGGAAGAC'TTCAAGGAGGTCCTAAGGGTGCGATTCGCTACTCTCTA 10 GT1TTTGCTGCTGTAGGCACAGCATTTGATTATGCTACCCTTAAAGGAAAA CCAATGTTAGAGAGCTACCGTAACATGGAGTCATTCAAGTTACCTGAATG GTCTCCTATTAAAGTCCTCGACGAAGAAGCCTTAGCAAAGAAGAAAGCTC ATGAAGAGAAGATATTCCCTGAAAGAGTCCTCGGCAAATTGAACAAAGA ATAGTCTTAACCAACTAAGATTAT1TCTCTTTGCCCCCATAAATTFCTTA 15 AGTTGGAATlTGTTTATCGGTGATGTTTCGTGAAAGACTGAAAGTAATTC CAGACCTTGTAGATGAGACTTGAGGAGGA=TTTGGTTTTTTGTTGTITCCT CAAGGTAAAAAT=TTCTTGAGACATAAGAAAACATCTTrGTATGCTGACCT ACCCATAAAGCGTATATATTCATGGTTAATTATGGGCTTA 20 Nucleic acid sequence of pk3lI (SEQ ID NO:3) GTTTTTCTCCACAAGGTITF[CTGCAAACATGTCTGAACTTGCATTGTCAT CTCAAGAAGAGTCTCCAAGTAGTAATAAGATTGGTTTATCTTCTCTTCTTC TCTCTGACTTTCATCTCTTGCTCATTTATCCTCACTCACCCTTTCTATT TCTTACTTGCTTrCTTTTCACCTTACATCTTCAAGATCTCTCTTTCTTITC 25 ACCACTCTTTGTCACCACCACACTCTTGCTTCTTGCCTTATTGAGTACTT[A CATGTTCAAGACACTTGTCTCGACTCTGAATCACTAGAAACACAACCAAG CTTCCTTTTCTCCTTTGTAGTAAGCTTGGAAGTGTCTTGGAACACAAGTTT GATGTCAACAATGAGGGTTAAGTCATTGGAGGAGTTGGAAGCGTATAA GATGGTCGTCGAGGCTTGCTCGATGGAATGTGCGTCCGAGAATGAGATAT 30 GTTCGGATGAATTGACGTTTGTTGACAAATTCTGTAGCCATGAGAGCACG GTGTCGGAATCTTTGACCGATGAGACCCTTGAGGAGCAAGTTGAGATCCA ACCGTrGAAGTTTGAGGATGTGATTGTTTTGGAGAAAGAAGAAGAAACCA AGAAATGTGAAAAGGAAGAAGTAGAAGAACAAAAAGTCAAGCATAAAA GTGACGTTGTCCTCGATAACAGAGAAGAGCCGACAAAAGAAGAATCCAA 35 AGCTCAAAAAGTTGACCTTGTCGGAGATAGTAATAATGAAAGTTATGATC TCCCAAAACTGAGCAATTTTCTCGGAGAAGGAGAAGGTAAAAGAAATGTA GTGACTAAGAACGAAGAAGAAGATAATGTTTCTCTCCAAAGCTTrGGATC AATGAGAAAAGAGAAAGAATGGAGGAGAACATTGGCTTGCAAGCTATTT GAAGAACGACACAATGCTGACGTTGGACAAGGCATGGATCAGCTGTGGG 40 AGACTrACGAGACACAAACAGAGAAGAAGCAGCAAACCGAAGAAGAGA AGAAGAAGCTCAAGAAGAAGACGAAGTCGATGATGAAGACA AAGAGTAT AGAGAAGGAAGTTATAGTGGAGGAGGAAGATGATGATGGGATTGATCAT CAGCAACTTGTTGTTTACAAGCTTTGAAGTICTCAACAGGGAAGATGCAT TTGGGAATTGCGAGGCCTAACCTTTTGAAGCTATCTAAGGCTTTCAAAGGC 45 ATTGGACGTTTTTACAATGCTAACAAACATTCCAAGAAAGCTTGAAAAGG AGATGAATAATAAAACTTTGTATTAATTGGGATCTATAAACAATGTAACTT GTAAGTTTCCATTGTTTTGGGCAAGTTCTATGAACAATGTAAGGGAAAAT AAAAGGTAAAGGCTAGGATTGCCATATGTGTTTAGCTTTGATCTTAACT -60- TTCTTTCCTATCCTTGTATATITfGGGACGGATAACCCGTAATGGCCCGTA TCGATTGAG Nucleic acid sequence of pk312 (SEQ ID NO:4) 5 ACACTGTGAGATTCAAGTGTAAAGTGCTCTCTCCCCAATGGCTAATCACCA CCGACTTTTACGCGGCGGCGGATCTCCGGCCATAATCGGTGGCAGAATCA CACTCACAGCTTrCGCTTCCACTATCGCACTCTTCCTCTTCACTCTCTCCTT CTTCTrCGCTTCAGATTCTAACGATTCTCCTGATCTCCTTCTTCCCGGTGTT GAGTACTCTAATGGAGTCGGATCTAGAAGATCCATGTTGGATATCAAATC 10 GGATCCGCTTAAGCCACGGTTGATTCAGATCCGGAAACAAGCTGATGATC ATCGGTCATTAGCATTAGCTTATGCTTCTTACGCGAGAAAGCTTAAGCTCG AGAATTCGAAACTCGTCAGGATCTTCGCTGATCTTTCGAGGAATTACACG GATCTGATTAACAAACCGACGTATCGAGCTITGTATGATTCTGATGGAGCC TCGATTGAAGAATCTGTGCTTAGGCAATTTGAGAAAGAAGTTAAGGAACG 15 GATTAAAATGACTCGTCAAGTGATTGCTGAAGCTAAAGAGTCTTTTGATA ATCAGTTGAAGATrCAGAAGCTGAAAGATACGATTTTCGCTGTTAACGAA CAGTTAACTAATGCTAAGAAGCAAGGTGCGTTTTCGAGTTTGATCGCTGC GAAATCGATTCCGAAAGGATTGCATTGTCTTGCTATGAGGCTGATGGAAG AGAGGATTGCTCACCCTGAGAAGTATACTGATGAAGGGAAAGATAGACC 20 GCGGGAGCTCGAGGATCCGAATCTTACCATTACGCTATATTTTCGGATAA TGTGATTGCGGCTTCGGTGGTTGTGAACTCTGCTGTGAAGAATGCTAAGG AGCCGTGGAAGCATGTITrCACGTTGTGACTGATAAGATGAATCTTGGA GCTATGCAGGTTATGTTTAAACTGAAGGAGTATAA AGGAGCTCATGTAGA AGTTAAAGCTGTrGAGGATTATACGTTTITGAACTCTrCGTATGTGCCTGT 25 GTTGAAGCAGTTAGAATCTGCGAATCTTCAGAAGT'ITATTTCGAGAATAA GCTCGAGAATGCGACGAAAGATACCACGAATATGAAGTTCAGGAACCCC AAGTATTTATCTATATTGAATCACTrGAGGTTTTATTTACCCGAGATGTAC CCGAAACTACATAGGATACTGTTTGGACGATGATGTGGTTGTGCAGAA GGATTTAACGGGTCTGTGGGAGATTGATATGGATGGGAAAGTGAATGGAG 30 CTGTAGAGACTTGTTGGGTCGTTTCATCGGTACGCTCAATACATGAATT TCTCACATCCTTTGATCAAAGAGAAGTTAATCCCAAAGCATGTGCGTGG GCGTATGGAATGAACTTCTT-TGATCTTGATGCTTGGAGAAGAGAGAAGTG CACAGAAGAATATCACTACTGGCAAAATCTGAACGAGAACAGGGCTCTAT GGAAACTGGGGACGTTACCACCGGGACTGATCACCTTTTACTCAACCACA 35 AAGCCGCTGGACAAATCATGGCATGTGCTrGGGCTGGGTTACAATCCGAG CATTAGCATGGATGAGATCCGCAACGCTGCAGTGGTACACTTCAACGGTA ACATGAAGCCATGGCTTGACATAGCTATGAACCAGTrCGACCACTrTGG ACCAAACACGTCGACTATGACCTCGAGTTTGTTCAGGCTTGCAATTGGC CTCTGAACTATGAAAATTTTCTTTATCATCAAAATCTGAAAGCATATGTTG 40 TTTGTTACTTCAGCTCTACGAAGFTTAACCTTAGTTTTTGTTTGTGTTTAT TTATATATTITGGGGGTTAGTAGAACACTTGTATTTTGTTCATAGCTATC TTGTTCTATGGCAACCTATAATCAAAGCTTAATTATAAAGTCACATTATG cc 45 Nucleic acid sequence of pk3l3 (SEQ ID NO:5) TTITrCTTCTTCTTCCATITITGTTCTCACGTCGCTCTCTCTT-T1TCG AGATTCAGCTGTAAAACCCTAACTAGCGCCATAGCCAAGGAAGCTTTCCT CAGATCGTCTCTCCGAAATTCCGGTTAATCGTCAGTTAAGGGGAAAATT -61- AGGCTATGGCGATGTTAGGTGCACAGCAAGTTCCAGCAGCAGCTTGTACT CCAGATATGGTTGGGAATGCTTTGTGCCCCAGTATTATCACATATTGCAT CAATCACCTGAGCATGTTCACAGATTTACCAAGAGATTAGCAAGTTAGG TCGTCCTGAAGAGAATGGTTTAATGAGCATCACTTCTACCTTGCAAGCTAT 5 TGACAAGAAGATAATGGCGCTrGGTTACGGTGTAATCAGTGCAGAGATAG CTACTGTGGACACACAAGAATCTCATGGAGGTGGTTATATTGTACTGGTG ACTGGGTATTTGACGGGAAAAGACAGTGTCAGGAGGACGTTTAGTCAGAC CTTCTTCCTTGCTCCACAGGAGACAGGATACTTTGTCTrGAATGATATGTT TCGATTCATTGATGAAGGCACTGTCGTACATGGAAATCAGATTCCAGTGA 10 ACAACGTCCAAGCTCCTGTCAACACTACCAGGACACAGCTGCTGCGAAG GAAATTCCAGATGACTTTGTTCAGGAGAAATATGTCCAAGAGAATCATGC TGTTAAGCAAACCGAGGTGTTGTCCAAGAGCATTAATGAGCCTGAAAAAG TGT'TCACGCCCTCTGAAGATGAACAAGTATCAGCTGCAGAAGAAGCTCTG GTGACTGAAACAGTTAATGAAGCACCAATTGAAGTGCAAAAGGTTGGAG 15 AATCTGATTCTAGGACTGGCGAAATTCCAAAGAGATCTTATGCATCAATT GTGAAGGTTATGAAAGAAAATGCTGCACCAATGTCTGCTTCGAGAACTCC AACAAAGGTGGAACCAAAGAAACAAGAAGATCAAGCCATTCATATCCCT CTACCAACACCATTGTCTGAGAAATCAGATTCAGGAGCAAATGTTGCTGT AAATGAGAACAATCAAGAGAATGAAAGAGCTCTAGGTCCATCCATCTATC 20 TAAAGGGTTrACCCCTTGATGCAACACCTGCCTTGCTTGAGAATGAGTTCC AGAAATTT'GGACTTATTAGGACCAATGGAATTCAAGTGAGAAGCCAGAAG GGATTCTGTrGGTTTGTTGAGTTGAATCCGCAAGTTCCATGCAAAGC GCTATCGAGGCATCACCTGTCATGCTCAATGGACACAAAGTTGTTGTGGA GGAAAAGCGATCTACCGCAAGAGGGAACTATAGAGGACGTTCGACGTTTG 25 GTGTAAACACAGGCTACAGAAACGAAGGAGGAAGGGGTCGTGGGAGCTT TGGAGGTGGAAGAGGAGGATATGGCCGGACCGATTTCAACGGATATGGT AATAACAGGGGAAACAATAGAGGCGGATACGCAAACCGAGCAAATGGTG ATGGTGGTGGGTTCCCGAGGGCCAATGGTAACAATGGACGAGTAAGACGT GGTGGCGGAAATGATGCTAACAGAGCTACGAAACCCGTGGATGATGCTCC 30 CCGTGTGTCTGTTGCTGCGTAAATGTGCTTTTGAAACAAAAAGCTCTATTG GTTTAGAGAGTTrAGGCGTAGAGCAATGGCAAAAAAAAACACTATTATT TTCTTTCACTGTGTCGCCATTFTATTAATTGGAGTCAAAACTTGAGAGCA AGAGAGAGTTTCGTCGGTTCTTGCTTGTCTATITTCTTCACTGCTAATGA AATCTCTTTCTTCATGTGGCTC 35 Nucleic acid sequence of pk314 (SEQ ID NO:6) GAAAGAAATCAAATACCTTCAGATCTCTATCTrCCTCATTCACACACCCTC TCTCTCTTCTCCTTTTCTCTCTTCTCCTTTCTCTATCTCCCTCTTTGTTCCGT TCGCATCCTCTAATCATCGTCAACAAGCCGACGAAGAGAGAAACGAATCC 40 AAAGTTCGTTACTrGAAAGCTACCCAGAAGAATTCAAATCTCAGGTACTTT TCCTGTGGATTGATCTGGGCACTGCTTAT-TAGGGATTGATrGGATCTAC AAAATCTGCCTrCTGGGTGATITCAATTTCACGGAAATGGTGAGGATTAA GAAAGTAGAATCGTTCTACGCGAAGCTTCGTGAGTCAGCTACTTCATTATC TTCACAGAATCCACT=TTGATATTTCCTTCAACATCTGATGTTGATTCACTT 45 TGTGCGCTTAAGGTTATTACTCATATCCTTGAATCAGATTCGATFCAGTAT TCTTGTTTCCCTGTATCGTCTTTTTGGAGATTCACAAGTATGCTGGTCCTG CTGGTTTGTGTTCTACTrCGTTGGAGAGTCCTCCTGTTACTATACTGTTGAT TAATTGGGGTTGTCACCGTGATTTGAAGCTTGTGTrGAAGTTAGGTCCTTC GGCTCGTGTTFTCGTTGTTGATAGTCATAGGCCTATTCATTTGCATAATCT -62- AGTGATTATAATGAGCAAGTTGTTGTTCTTCATACTGATGATGATGAGAGG CAAGGTGATTrGGCTTATGATTrCGATGTGTTGAAATrGGCGAATGAGAG CTTTCAGTTACGTGTAGAAGATGCTGGTGAAGAATCTGATGAGGAGGAGG AAGATGAGGAAGAGGATGAGGAGGATGATGATGATGATGATGGTGATAG 5 GCCAAGTAAGAGGAGGAAAATGGGAGATGGTGTGAAGGTTTTCAAGAAG CTAAAGAGGGATTATTACAAGATGGGGACTTTTCATGGGAAGCCATCGGG GTGTTTGTTGTTTGAGCTATCTCATATGTTGAGGAAGAACACTAACGAGTT GTTGTGGCTGGCTrGTGTTTCTTTGACTGATCAGTTrGTTCATGAGAGGTT GACTGATGAAAGATATCAAGCTGCGGTTATGGAGCTTGAACAACACATCA 10 ATAGCTCAGGGAATATAGATAAGATCACTAGTGTTACTCTGAAAGATGGA ACCAAGGTTCGAGCACCAGACTGTTCAAGAATCTCTTATGAAGAAGAGCC TAGGCTTATGCT-TCTTAGAGAGTGGACGTTGTTTGACTCCATGCT-rTGT'rCT TCATACATTGCGACTAAGTTGAAGACATGGAGTGATAACGGTATCAAGAA ACTTAAGCTTCTTCTAGCGCGTATGGGATTTGCACTTATCGAGTGTCAGCA 15 AAAG2TCCGTACATGAGCCTTGAGGTGAAGAGGAAGATGAAGCAAGAG TTTGATCGGTTTTTGCCAGAATATGGGCTrAATGATTTCTACTACCGGAGT TTCTTGCGGCTTCATGGTTATAGCTCAAGGGTCTCTGCTGCAGATGTTGTC TATGGTATTACAGCACTTCTTGAATCATTTCTTGGGTCAGGTGGCTCCTCT GCTTCAAAACAGTTTGGTGAAGCTrATGATGCTCTGTCTGAACAATTTG 20 GATAAACTTCGATCTGGGATGCAACAAGCAATCAAGGTTCAACGAGCAAT TCTTAGACAAGGAAGTGCAGCAATCACTAAAAGTGGATGCATTCGAAGTG GTAGGAAATTCAGATGGGTAAAGATTGAAGATTCAATGGATGCGAAGTAT TTGGGATATCCTCAGGCCTTAACAAAATTCTGTTACTTTCTGATGGATGCT TTGAGAGAGAAAGGAGCTAGGATGAAACCAATGCTATGTGCCTGCGCATC 25 TCAACAACCTGGGAAGATACTCGTGGT-TGGGGTTTGTGGGAAACCGAGGC TCGGGGCAGTCAGAGGGAATGCTTTGGCAATGCTTTCAGAAAGGCAGCT CAAGAAAGTAGAGCTGATTACTTTCACGAGCTATTCGAGTCTTCTTGGATT GTCTTGGATGCTTCTGCAGTTAACTCTTTCATGATTAGATTAACCGAGAAG CTCTGACATAGTCTCATTGTTCTTCGATTCAGTGTGTITCTTTTATAGTTT 30 TCAGTTATCTCACTGTTTGCATITrACGAGCCTGTGTAATAGGCACA ATCTGTTATCAATCATGTAACTTGTrTAAT Nucleic acid sequence of pk315 (SEQ ID NO:7) ATGCAAATAGGTCAAGCCTTAGCCGCAGCAAAGGAAGGTGAGTCTCAGAT 35 GATCGTGATGATGGGTAACAATCTTTCTTTAACAAGCATTATTCTCAATGG AGATCCATCTATAGAGCATAAAGGAAAACTTACTTGCCTTGACGAACAAG TCAAGATATCTCAGTTCTATCACTGCGAGGGCTrACTGCTATGCATTTTAA AAGATGATTCTAGGTTTGTGGTTTGTAATCCGTATTTGGAGCAAACAAGGT GGATCGAACCAAGATATTCCCATCGTCCATACGGAATGGATAGGTTCTCTT 40 ACGCTCTTGGATACGTGAATACGGATTCTTGTCGTAGCTACAAGTTGTTGA GGTTTATAGATTATTACTACAATGCACCCGAGAAGCAATTCTTTTGGTATG AAGTCTACGATTTTGACTCTGATTTATGGACTACTCTTGATGTCACTCCAC ATTGGCGTATAGCGTTTTGTAACACTGGCGTTCCTTAAGGGAAACACTT ACTGGTGTGCTGCAGAAAGGAACGTAGATGTAGATGAAGTCTTAGCTAAT 45 CGCTTAATCTGTTTTGATTTACAAAAGAGAAGCTTGCGG=TTACTTCAG CACGATGAATCAAATCCATATGAGCTTGACTTGTGGATTACAACTAAGAT TGAGACAGAAGAGGTGTTGTGGAGCAAGTTCTTGAGAGTGGAAACAGCTG GTTTTAATAGTTATGTTCCTTFTATAAGTGGAAGTTCTTCATTGACGAGG AGAAGAAAGTCGCCTTTGGTTTTGATGAACGTAACCGCCAGAGAGTTATT -63- GTCATTGGAGAGGCTGGATACTTGAGGGGATTGGATCTCGTTGGGGATTT TGGAGACCAAAGCTGTAAGCCAGATCTATGCTCTTATGTTCCAAGTTTAGT GCAAATCAAGCAACCTGAAGGAGGGGAAAGGGAAGAAGAAAGCGAATAT GGAGAAGCTTCGATATGA 5 Nucleic acid sequence of pk316 (SEQ ID NO:8) GTCTCTCCTCTGCATCTCCTCTGTTCCTCAGGTTTCTCTGCTCATGGCTGCT TATGGTCAAATCTCCTCGGGAATGACTGTAGATCCTCAGGTFCTCTCTTCC TCCAGAAACATTGGAGTTTCCCTATCACCTCTCCGGAGAACACTAATCGGC 10 GCCGGAGTTAGGTCTACTAGTATCTCTCTCCGTCAATGTTCTCTCTCCGTTA GATCGATrAAAATCTCCGAAGATAGCCGCAAACCTAAAGCTTATGCAGAG AACGGTGCTTTTGATGTGGGAGTITGGATTCTTCATCATATAGATTGGCT GATrCAAGAACAAGTAGTAATGATTCAAGGAGGAAGACTAAGATTGTGTG TACGATTGGACCGTCTTCGAGTTCTAGGGAAATGATTTGGAAACTCGCGG 15 AAGCTGGAATGAATGTGGCTCG1TFGAATATGTCTCATGGTGATCATGCTT CTCATCAGATAACTATTGATTAGTrAAGGAGTATAA TTCTTTGTTTGTTG ACAAAGCTATTGCTATTATGTTGGATACAAAGGGTCCTGAGGT-FCGAAGC GGGGATGTACCGCAGCCGATATTTCTTGAAGAGGGTCAAGAGTTrAACTT TACTATCAAGAGAGGTGTTTCGCTTAAAGACACTGTTAGTGTAAATTATGA 20 TGATTGTGAACGATGTTGAAGTrGGGGATATACTTTTGGTGGATGGTGG AATGATGTCGTTAGCTGTTAAATCAAAGACGAGTGATTTGGTGAAGTGTG TGGTTATTGATGGTGGAGAGCTTCAATCTAGACGTCACTTGAATGTTCGAG GAAAGAGTGCGACTCTTCCATCCATTACAGACAAAGATTGGGAAGACATA AAATTTGGAGTGGACAACCAAGTCGATTrCTACGCCGTCTCCTTTGTfAAG 25 GATGCTAAAGTTGTCCATGAGTTGAAGAACTATCTCAAAACCTGCAGTGC AGACATATCGGTGATTGTGAAAATGAAAGTGCAGACTCTATAAAGAATC TTCCTTCTATCATATCTGCTTGTGATGGGGCAATGGTTGCTCGTGGAGATC TTGGAGCTGAACTTCCCATTGAAGAGGTCCCGTTGTTACAGGAAGAAATA ATCAGAAGGTGTAGAAGCATTCATAAACCAGTGATTGTTGCCACAAACAT 30 GCTAGAGAGTATGATTAATCATCCAACGCCTACAAGAGCTGAAGTCTCTG ACATTGCAATTGCAGTACGTGAAGGCGCAGATGCTATCATGCTTTCTGGTG AAACCGCACATGGAAAGTTTCCGCTGAAAGCTGTTAACGTAATGCATACT GTGGCGTTGAGAACCGAGGCAAGTCTACCTGTCAGAACCTCGGCATCCCG TACCACTGCTTACAAGGGTCACATGGGCCAAATGTTrGCTTTTCATGCTTC 35 TATAATGGCAAATACACTGAGCTCACCGCTAATTGTATTTACGAGAACCG GATCCATGGCAGTGCTTCTAAGCCACTACCGCCCATCTGCAACAATTTTCG CCTTCACAAACCAGAGAAGAATAATGCAAAGGCTTGCTCTTTATCAAGGT GTCATGCCTATATATATGGAGTTCTCGGATGATGCAGAAGATACATATGC CCGGTCTCTCAAACTCTFACAGGACGAGAATATGCTCAAGGAAGGACAAC 40 ATGTAACTCTTGTCCAAAGTGGCTCGCAACCCATITGGCGTGAAGAATCA ACACATCTCATACAAGTCCGTAAGATAAAGATAGGTGGATGATGTT--A CTTCTTGAGCTACACAACATCTTGTTACTCAGCT-TCTTTCTCTTACAC AGT7CGATCCATAT 1-1I GAATCACTCACAGTGAATCAAACAACCATAT AAAATTTTAAGTTATTGAAGCT1T11rCTGGTTATAG 45 Nucleic acid sequence of pk317 (SEQ ID NO:9) AGAAAAAAAAAAAAAAATCCAAATTCAACACTCTCACACTTrCGATATCTC CGCCTTCATTCTCCTCAGAGCCAACTGTCCTGAGATTCGATTTCGATTTCT -64- CCGATCTCTCTTCCTCCGTCGCCGGCGAAACCATGTCTCAGTCTATTCAAT TCTCCACTCCTTCACACACTCCTCACCTTCTCCATCTCCCTCACTCACAATT CAACCGTCCTCTCTCCTCTATCTCCTTCCGTCGCTTCCCTCTAACAACCATC AAATACACTITCCATCAGAGCCTCCTCGTCATCATCTCCTTCACCGGATCTC 5 GATTCATCGTCCTCATCATCATCCTCGCAAGTACTTCTCTCACCTAACGGT ACTGGTGCTGTGAAGTCTGATGAGAGATCCGTTGTCGCTACGGCGGTTAC GACTGATACGTCTGGGATTGAGGTTGATACTGTGACGGAAGCTGAGCTTA AGGAGAATGGATTTAGAAGTACGAGGAGGACGAAGCTGATCTGTACGAT CGGACCGGCGACTTGTGGATTTGAGCAGCTTGAGGCGCTTGCTGTGGGAG 10 GTATGAATGTGGCAAGGCTTAATATGTGTCACGGTACGCGTGATTGGCAC CGCGGTGTGATTCGTAGTGTTCGGAGGCTTAATGAGGAGAAAGGCTTTGC GGTTGCTATTATGATGGATACTGAAGGTAGTGAGATTCATATGGGAGATC TTGGTGGTGAAGCTCAGCTAAAGCAGAGGATGGTGAGGTTTGGACTTTC ACTGTTAGAGCTITGATTCTTCTCGTCCTGAACGTACCAT-TAGTGTTAGCT 15 ACGATGGTTTCGCTGAAGATGTAAGAGTTGGGGATGAACTTTTGGTrGAT GGTGGGATGGTGAGATTTGAAGTGATTGAGAAGATTGGTCCTGATGTTAA GTGTCTATGTACCGATCCTGGATrGTTGCTTCCTCGAGCTAACTTGACGT7 TTGGAGAGATGGAAGTCTTGTACGAGAGCGTAATGCCATGCTTCCAACAA TTC'TTCCAAGGACTGGTrGGATATTGATTTTGGAATrGCTGAAGGTGTGG 20 ATTCATrGCTGTATCGTTTGTCAAGTCGGCTGAAGTCATTrAATCACCTEA AAAGTTATCTTGCTGCTCGTrCCCGTGGAGGGGAAATTGGAGTGATTGCA AAGATCGAGAGTATCGATTCACTGACCAAT-rTGGAAGAAATTATTCTAGC ATCAGATGGGGCCATGGTTGCAAGAGGAGATCTGGGAGCTCAGATACCTC TTGAGCAAGTTCCAGCAGCTCAACAGAGAATCGTCCAAGTATGCAGAGCT 25 CTTAACAAACCCGTCATTGTCGCTTrCACAGCTATTGGAGTCCATGATTGAG TACCCAACTCCAACCAGAGCAGAAGTTGCCGACGTGTCTGAAGCAGTAAG ACAAAGATCAGATGCATTGATGCTCTCTGGAGAATCAGCTATGGGACAAT TCCCAGACAAGGCGCTCACGGTTCTAAGGACTGTCAGTTTAAGAATCGAG AGATGGTGGAGGGAAGAGAAACGCCATGAGTCTGTACCGCTTCAAGCCAT 30 AGGCTCTTCATTCAGACAAAATCTCAGAAGAGATCTGTAACTCAGCTGC TAAAATGGCTAACAATCTTGGAGTGGACGCGGTTTCGTTTACACAACGA GCGGACACATGGCATCACTGGTCTCCCGATGTCGCCCGGACTGCCCGATC TTTGCTTCACAACCACAACCTCAGTGAGAAGACGCTTAAACCTACAATG GGGACTTATCCCATTCCGTCTCAGCTTCTCAGACGACATGGAAAGCAACTT 35 GAACAAAACATTCTCGTTACTGAAATCAAGAGGTATGATCAAATCTGGTG ACCTCGTGATCGCAGTCTCGGACATGCTGCAATCAATCCAGGTAATGAAC GTCCCGTAATTCTCTCTCTTTATACAATTTCGCAATCCCGCAA AAGAGTGT TTTGTTTCCTACTTTTGTTACTGTTTAGACTACTCTTACATTAGAFTTCCA GAGGCATCATCATCTTCGGTTTGTTAACAACAGTAATGTGTAAGCTTTGTF 40 TGTAGTGTGTACTGTTTGTTTTTGGTTTTCAATAATATCAGTAATCTTATTC AAATATFCGATTCTATC Nucleic acid sequence of pk318 (SEQ ID NO:1o) ATGGGTAAAGTAGCTGTTGGAGCGACTGTTGTTTGCACGGCGGCGGTTTG 45 TGCGGTGGCTGTTGGTTGTTCGACGACGGATGCAGAGCTCAGGGAAGT GGGGACGTGTTTTGGCTATCCTCAAGGCCTTTGAAGAGGA TTGTGCGACTC CGATCTCGAAACTGAGACAAGTGGCTGATGCTATGACCGTTGAGATGCAT GCTGGTCTTGCATCCGACGGTGGTAGCAAACTCAAGATGCTTATCAGCTA CGTTGATAATCTTCCTTCCGGGGATGAAAAGGGTCTCTTTTATGCATTGGA -65- CCTAGGGGGGACAAACTTCCGTGTCATGCGTGTGCTTCTTGGCGGGAAGC AAGAGCGTGTTGTAAACAAGAATTCGAAGAAGTTTCGATTCCTCCTCATr TGATGACTGGTGGTTCAGATGAGTTGTTCAATTATAGCTGAAGCTCTTG CGAAGTTTGTCGCTACAGAATGCGAAGACTTTCATCTrCCAGAAGGTAGA 5 CAGAGGGAATrAGGTTTCACTTTCTCGTTTCCTGTTAAGCAGACTTCTCTG TCCTCTGGTAGTCTCATCAAATGGACAAAAGGCTTTTCCATCGAAGAAGC AGTTGGACAAGATGTTGTTGGAGCACTTAATAAGGCTCTGGAAAGAGTTG GTCTTGACATGCGAATCGCAGCACTTGTTAATGATACCGTTGGAACACTA GCCGGTGGTAGATACTATAACCCGGATGTTGTTGCTGCTGTTATTAGGC 10 ACTGGGACAAACGCAGCCTATGTTGAGCGTGCAACCGCGATCCCTAAATG GCATGGTCTGCTTCCAAAATCAGGAGAAATGGTTATAAACATGGAATGGG GAAACTTCAGGTCATCACATCTTCCATTAACCGAGTTTGATCACACGCTGG ATTTCGAGAGTCTGAATCCAGGCGAACAGATTCTTGAGAAAATCATTTCC GGTATGTACTTGGGAGAGA1TTGCGAAGAGTTCTTCTAAAGATGGCTGA 15 AGATGCTGCTTTCT'TTGGCGATACAGTCCCATCTAAGCTGAGAATACCATT CATCATTAGGACTCCTCACATGTCGGCTATGCACAACGACACTTCTCCAGA CTTGAAGATTGTTGGGAGCAAGATTAAGGATATATTGGAGGTCCCTACAA CTTCTCTGAAAATGAGAAAAGTTGTGATCAGTCTCTGCAACATCATAGCA ACCCGAGGAGCTCGTCTCTCTGCTGCTGGAATCTATGGTATTCTGAAGAAA 20 CTGGGAAGAGATACTACTAAAGACGAGGAGGTGCAGAAATCGGTTATAG CCATGGATGGTGGATTGTTTGAGCATTACACTCAGTrTAGTGAGTGTATGG AGAGCTCACTAAAAGAGTTGCTTGGAGATGAAGCTTCAGGAAGCGTTGAA GTCACTCACTCCAATGATGGATCAGGCATTGGAGCTGCGCTTCT-FGCTGCT TCTCACTCTCTCTACCTTGAAGACTCTrAA 25 Nucleic acid sequence of pk319 (SEQ ID NO: 11) AAGCACTTCTTCTCCGCCTTCGTAAGTTCCGCCGAAAAGAACCAAATCCTT CACTACTCTGTCTCAGCTTTCGACCTCTCTCTrCTCATTCCTTTGCAACTTC TCACTTCTCGAATTCCTTCTCTTCAAAATCAGAAATGGCTCAAGTGGTTGC 30 TACCAGGTCAATTCAAGGCTCGATGTTATCTCCCAACGGTGGATCTGTGTC TACAAGATCCGAGAAGCTATTGAAACCAGCGAGTTTGCAGTGAAGGTTC TTGGCAACGAAGCAAAGAGAAGTGGAAGAGTCTCTGTAAGAAGCAGAAG AGTGGTTGATACTACTGTGAGATCCGCTCGTGTTGAGACTGAAGTCATCC TGTTTCTCCTGAAGATGTGCCTAACAGAGAGGAGCAGCTTGAGAGGTTGT 35 TGGAAATGCAGCAGTTGGTGATACATCGGTAGGGATGTGGTCGAAGCCG ACAGTGAGGAGGAAGACAAAGArGTTTGCACCGTTGGTCCGTCGACCAA CACACGAGAAATGATATGGAAATrGGCTGAAGCTGGGATGAATGTTGCTA GGATGAATATGTCTCATGGAGATCATGCTTCACATAAGAAGGTTATTGATT TGGTTAAAGAATACAATGCACAAACTAAAGACAACACTATTGCTATCATG 40 CTTGACACCAAGGGTCCGGAAGTTAGGAGTGGAGATTTACCTCAGCCAAT TATGTTAGATCCTGGTCAAGAGTTTACCTTTACAATTGAGAGAGGAGTCA GCACACCAAGTTGTGTCAGTGTTAACTATGATGATTTCGTTAATGACGTGG AAGCGGGTGACATGCTTCTTGTTGATGGTGGTATGATGTCGTTTATGGTGA AGTCAAAGACCAAAGACTCTGTCAAATGTGAAGTTGTTGATGGTGGAGAA 45 CTTAAGTCAAGGAGACACCTGAATGTCCGAGGAAAGAGTGCAACTTTACC TTCAATCACTGAGAAGGACTGGGAGGATATTAAAmTTGGAGTGGAGAACA AAGTTGACTTTATGCAGTTrCCTTTGTCAAAGATGCTCAAGTTGTACACG AGTTGAAGAAATACCTFCAAAATAGTGGTGCTGATATACACGTGATAGTG AAAATTGAGAGTGCAGACTCCATACCTAACTTGCACTCCATATCACAGC -66- ATCAGATGGGGCAATGGTTGCAAGAGGTGATCTTGGTGCAGAGCTTCCAA TTGAAGAAGTCCCCATTCTTCAGGAGGAGATCATTAACCTGTGCCGTAGT ATGGGAAAAGCTGTTATTGTTGCGACTAACATGCTTGAGAGTATGATAGT TCATCCAACTCCAACCCGGGCAGAGGTCTCAGACATTGCTATCGCTGTTAG 5 AGAAGGTGCTGATGCGGTAATGCTCAGGAGAAACTGCTCACGGAAAGT TCCCATTGAAAGCTGCTGGAGTGATGCACACTGTTGCATTGCGAACAGAA GCAACCATTACTAGCGGTGAAATGCCACCTAATCTTGGTCAAGCCTTCAA GAACCATATGAGTGAGATGTTTGCATACCATGCAACCATGATGTCAAACA CACTTGGAACTTCAACTGTTGTCTTCACCAGAACCGGTTTCATGGCCATAT 10 TGTTAAGTCACTATCGTCCTTCCGGCACAATCTATGCCTrCACAAATGAGA AAAAAATACAACAAAGATTAGCTTTGTATCAAGGTGTATGCCCCATATAT ATGGAGTTCACAGATGATGCAGAAGAAACTTGCTAATGCTTTGGCTAC ATTACTGAAACAAGGAATGGTGAAGAAGGGAGAGGAAATAGCAATCGTA CAGAGCGGTACACAGCCAATCTGGCGATCTCAATCGACACATAACATCCA 15 AGTCCGCAAGGTTrAAAGCTTCTTTAAGATGGGATGTCTTTAATATGTAG AACCTCGTTTTTGGTTATAATTTCGTTGCATGTCTCTCTTCTCTTGTACTA TTCACACTTGTTGTTTGCTGTATCTTCTTCTTCAGTTTGCTTTGCTACGATT GTGGTTTGGAGACATTATAGCTCATTAACTGTTTGTGAGACCAAATGTG TCAGAATCCGCTATT 20 -67-

Claims (26)

1. An isolated Lipid Metabolism Protein (LMP) nucleic acid comprising a polynucleotide sequence selected from the group consisting of: a) a polynucleotide sequence as defined in SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11; b) a polynucleotide sequence encoding a polypeptide that is encoded by a polynucleotide sequence as defined in SEQ ID NO:I, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11; c) a polynucleotide sequence having at least 70% sequence identity with the full-length LMP nucleic acid of a) or b) above; d) a polynucleotide sequence that is complementary to the full-length LMP nucleic acid of a) or b) above; and e) a polynucleotide sequence that hybridizes under stringent conditions to the full-length LMP nucleic acid of a) or b) above.

2. The isolated LMP nucleic acid of Claim 1, wherein the polynucleotide sequence encodes the polypeptide that is encoded by a polynucleotide sequence as defined in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11.

3. The isolated LMP nucleic acid of Claim 1, wherein the polynucleotide sequence is defined in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11.

4. The isolated LMP nucleic acid of Claim 1, wherein the polynucleotide sequence has at least 90% sequence identity with the full-length LMP nucleic acid of a) or b) of Claim 1, and wherein the isolated LMP nucleic acid encodes a polypeptide that functions as a modulator of a seed storage compound in a plant. -68-

5. The isolated LMP nucleic acid of Claim 1, wherein the polynucleotide sequence is complementary to the full-length LMP nucleic acid of a) or b) of Claim 1, and wherein the isolated LMP nucleic acid encodes a polypeptide that functions as a modulator of a seed storage compound in a plant.

6. The isolated LMP nucleic acid of Claim 1, wherein the polynucleotide sequence hybridizes under stringent conditions to the LMP nucleic acid of a) or b) of Claim 1, and wherein the isolated LMP nucleic acid encodes a polypeptide that functions as a modulator of a seed storage compound in a plant.

7. The isolated LMP nucleic acid of Claim 1, wherein the nucleic acid is located in an expression vector.

8. The expression vector of Claim 7, wherein the LMP nucleic acid is operatively linked to a heterologous promoter selected from the group consisting of a seed-specific promoter, a root-specific promoter, and a non-tissue-specific promoter.

9. A method of producing a transgenic plant having a modified level of a seed storage compound comprising, transforming a plant cell with an expression vector comprising a lipid metabolism protein (LMP) nucleic acid and generating from the plant cell the transgenic plant, wherein the nucleic acid encodes a polypeptide that functions as a modulator of a seed storage compound in the plant, and wherein the nucleic acid comprises a polynucleotide sequence selected from the group consisting of: a) a polynucleotide sequence as defined in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ [D NO: 11; b) a polynucleotide sequence encoding a polypeptide that is encoded by a polynucleotide sequence as defined in SEQ ID NO: 1, SEQ 1D NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11; c) a polynucleotide sequence having at least 70% sequence identity with the full-length LMP nucleic acid of a) or b) above; -69- d) a polynucleotide sequence that is complementary to the full-length LMP nucleic acid of a) or b) above; and e) a polynucleotide sequence that hybridizes under stringent conditions to the full-length LMP nucleic acid of a) or b) above.

10. The method of Claim 9, wherein the LMP nucleic acid comprises the polynucleotide sequence of SEQ ID NO:l, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11.

11. The method of Claim 9, wherein the LMP nucleic acid comprises a polynucleotide sequence encoding the polypeptide that is encoded by a polynucleotide sequence as defined in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, or SEQ ID NO:11.

12. The method of Claim 9, wherein the LMP nucleic acid comprises a polynucleotide sequence having at least 90% sequence identity with the polynucleotide sequence of a) or b) of Claim 9.

13. The method of Claim 9, wherein the LMP nucleic acid hybridizes under stringent conditions to the LMP nucleic acid of a) or b) of Claim 9.

14. The method of Claim 9, wherein the LMP nucleic acid comprises a polynucleotide sequence complementary to the LMP nucleic acid of a) or b) of Claim 9.

15. The method of Claim 9, wherein the level of total oil content in a seed is modified.

16. The method of Claim 9, wherein the level of a seed storage compound is increased in the transgenic plant as compared to an untransformed wild type variety of the plant. -70-

17. The method of Claim 9, wherein the LMP nucleic acid is operatively linked to a heterologous promoter selected from the group consisting of a seed-specific promoter, a root-specific promoter, and a non-tissue-specific promoter.

18. A method of modulating the level of a seed storage compound in a plant comprising, modifying the expression of a Lipid Metabolism Protein (LMP) nucleic acid in the plant, wherein the LMP nucleic acid comprises a polynucleotide sequence selected from the group consisting of: a) a polynucleotide sequence as defined in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11; b) a polynucleotide sequence encoding a polypeptide that is encoded by a polynucleotide sequence as defined in SEQ ID NO:I, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ [D NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11; c) a polynucleotide sequence having at least 70% sequence identity with the full-length LMP nucleic acid of a) or b) above; d) a polynucleotide sequence that is complementary to the full-length LMP nucleic acid of a) or b) above; and e) a polynucleotide sequence that hybridizes under stringent conditions to the full-length LMP nucleic acid of a) or b) above.

19. The method of Claim 18, wherein the level of total oil content in a seed is modified.

20. A transgenic plant made by a method comprising, transforming a plant cell with an expression vector comprising a lipid metabolism protein (LMP) nucleic acid, and generating from the plant cell the transgenic plant, wherein the nucleic acid encodes a polypeptide that functions as a modulator of a seed storage compound in the plant, and wherein the nucleic acid comprises a polynucleotide sequence selected from the group consisting of: a) a polynucleotide sequence as defined in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11; -71- b) a polynucleotide sequence encoding a polypeptide that is encoded by a polynucleotide sequence as defined in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO: 10, or SEQ ID NO: 11; c) a polynucleotide sequence having at least 70% sequence identity with the full-length LMP nucleic acid of a) or b) above; d) a polynucleotide sequence that is complementary to the full-length LMP nucleic acid of a) or b) above; and e) a polynucleotide sequence that hybridizes under stringent conditions to the full-length LMP nucleic acid of a) or b) above.

21. The transgenic plant of Claim 20, wherein the level of total oil content in a seed is modified.

22. The transgenic plant of Claim 20, wherein the plant is a dicotyledonous plant.

23. The transgenic plant of Claim 20, wherein the plant is a monocotyledonous plant.

24. The transgenic plant of Claim 20, wherein the level of the seed storage compound is increased in the transgenic plant as compared to an untransformed wild type variety of the plant.

25. The transgenic plant of Claim 20, wherein the plant is a high oil producing plant.

26. The transgenic plant of Claim 25, wherein the high oil producing plant is selected from the group consisting of rapeseed, canola, linseed, soybean, sunflower, maize, oat, rye, barley, wheat, pepper, tagetes, cotton, oil palm, coconut palm, flax, castor, and peanut. -72-