MXPA00011975A - Genes for desaturases to alter lipid profiles in corn - Google Patents

Abstract

The preparation and use of nucleic acid fragments comprising all or substantially all of a corn oleosin promoter, a stearoyl-ACP desaturase and a delta-12 desaturase which can be used individually or in combination to modify the lipid profile of corn are described. Chimeric genes incorporating such nucleic acid fragments and suitable regulatory sequences can be used to create transgenic corn plants having altered lipid profiles.

Description

GENES FOR DESATURASES TO ALTER LIPID PROFILES IN MAIZE FIELD OF THE INVENTION The invention relates to the preparation and use of nucleic acid fragments comprising all or substantially all of the promoter of a maize oleosin, a stearoyl desaturase ACP and a delta-12 desaturase, which can be used individually or in combination to modify the lipid profile of the grain. Chimeric genes comprising such nucleic acid fragments and suitable regulatory sequences can be used to create transgenic corn plants having altered lipid profiles.

BACKGROUND OF THE INVENTION Plant lipids have a variety of industrial and nutritional uses and are central to the function of the plant membrane and climate adaptation. These lipids represent a vast array of chemical structures, and these structures determine the physiological and Industrial REF.124739 lipid. Many of these structures result either directly or indirectly from the metabolic processes that alter the degree of lipid unsaturation. Different metabolic regimes in different plants produce these altered lipids, and their domestication of exotic plant species or modification of agronomically adapted species is usually required to produce economically large quantities of the desired lipid. The lipids of plants find their greatest use as edible oils in the form of triacylglycerols. The specific performance and healthy attributes of edible oils are largely determined by their fatty acid composition. Most vegetable oils derived from commercial varieties of plants are composed mainly of palmitic (16: 0), stearic acids (18: 0), oleic (18: 1), linoleic (18: 2) and linolenic (18: 3). The palmitic and stearic acids are, respectively, saturated fatty acids of 16 and 18 long carbons. The oleic, linoleic and linolenic acids are unsaturated fatty acids of 18 carbons, containing one, two and three double bonds, respectively. Oleic acid refers as such to a monounsaturated fatty acid, while linoleic and linolenic acids refer as such to polyunsaturated fatty acids. The relative amounts of saturated and unsaturated fatty acids in commonly used vegetable oils are summarized below (Table 1): TABLE 1 Acid Percentages Saturated and Unsaturated Fats in Selected Oils Cultivation Oils Saturated Mono-unsaturated unsaturated Cañola 6% 58% 36% Oil of 15% 24% 61% Soybean Maize 13% 25% 62% Peanut 18% 48% 34% Safflower 9% 13% 78% Sunflower 9% 41% 51% Cotton 30% 19% 51% Corn oil is comprised mainly of fatty acids from carbon chains still numbered. The distribution of fatty acids in typical corn oil is approximately 12% palmitic acid (16: 0), 2% stearic acid (18: 0), 25% oleic acid (18: 1), 60% linoleic acid (18: 2), and 1% linolenic acid (18: 3). Stearic and palmitic acids refer to saturated fatty acids, because their carbon chains contain only single bonds and the carbon chain is "saturated" with hydrogen atoms. The oleic, linoleic and linolenic acids contain one, two and three double bonds, respectively, and are referred to as unsaturated fatty acids. The fatty acids in corn oil almost always occur esterified to the hydroxyl groups of glycerol, thus forming triglycerides. Approximately 99% of the refined corn oil is made from triglycerides ("Corn Oil", Corn Refiners Association, Inc.m 1001 Connecticut Ave., N.W., Washington, DC 20036, 1986, 24 pp). Many recent research efforts have examined the role of saturated and unsaturated fatty acids in reducing the risk of coronary heart disease. In the past, it was believed that monounsaturates, in contrast to saturated and polyunsaturated ones, had no effect on serum cholesterol and the risk of coronary heart disease. Several recent human clinical studies suggest that diets high in monounsaturated fats and baas in unsaturated fats can reduce "bad" cholesterol (low density lipoprotein), while maintaining "good" cholesterol (high density lipoprotein) (Mattson et al. (1985) Journal of Lipi d Research 26: 194-202). A vegetable protein low in total saturation and high in mono-unsaturated can provide significant health benefits to consumers as well as economic benefits to the oil processors. As an example, canola oil is considered a very healthy oil. However, in use, the high level of polyunsaturated fatty acids in canola oil produces unstable oil, easily oxidized, and susceptible to the development of unpleasant odors and flavors (Gailliard (1980) in The Biochemistry of Plants Vol. 4, pp. 85-116, Stumpf, PK, ed., Academic Press, New York). Polyunsaturated levels can be reduced by nitrogenation, but the cost of this process and the concomitant production of the nutritionally questionable trans-isomers of the remaining unsaturated fatty acids reduce the overall quality of the hydrogenated oil (Mensink et al. (1990) N. Eng J. Med.? 323: 439-445). When exposed to air, the unsaturated fatty acids are subjected to oxidation, which causes the oil to have a rancid odor. Oxidation is accelerated at high temperatures, such as under frying conditions. The oxidation rate increases in the case of oils that contain higher degrees of unsaturation. In addition, linoleic acid with two double bonds is more unstable than oleic acid which has only one double bond. Oxidation reduces the half-life of products containing corn oil because it is the highest proportion of linoleic acid oil. Corn oil and products containing corn oil are often packaged under nitrogen in special packaging materials such as plastics or laminated sheets, or stored under refrigeration to extend shelf life. These extra measurements reduce oxidation and subsequently, rancidity adds a considerable cost to products containing corn oil. Other measurements to reduce the effects of oxidation in corn oil is to chemically hydrogenate the oil. This commercially important process by which hydrogen is added to the double bonds of unsaturated fatty acids changes the physical properties of the oil and extends the shelf life of products containing corn oil. Hydrogenated vegetable oils are used to make margarine, salad dressing, cooking oil, and shortening to mix, for example. Approximately half a trillion pounds, or approximately 40-50%, of corn oil produced in the United States is used for cooking and as salad oils (Fitch, B., (1985) JAOCS, Vol. 2, no. 11, pp. 1524-31). The production of a more stable oil by genetic means could clearly have value by reducing or eliminating the time and costs of entry of chemical hydrogenation. In addition to the economic factors associated with the chemical hydrogenation of corn oil, there are human health factors that favor the production of a high natural oleic oil. During the hydrogenation process, the double bonds in the fatty acids are completely hydrogenated or are converted from the ci s configuration to the trans configuration. The double bonds ci s cause a fatty acid molecule to "bend" which imparts crystallization and keeps the liquid oil at room temperature. During hydrogenation, the bonds ci s are directed in the trans configuration, causing the oil to harden at room temperature. Recent studies on the effect of dietary trans fatty acids on cholesterol levels showed that the trans isomer of oleic acid raises blood cholesterol levels at least as much as saturated fatty acids, which are known to some time, they raised cholesterol in humans (Mensink, RP and BK Katan, (1990), N. Engl. J. Med., 323: 439-45). In addition, these studies show that the level of undesirable low density lipoprotein is increased, and the level of desirable low density lipoprotein decreases in response to high diets in trans fatty acids. Large amounts of fatty acids are found in margarines, kneading butter, and oils used for frying; The most abundant trans fatty acid in the human diet is the trans isomer of oleic acid, elaidic acid. While oils with low levels of unsaturated fatty acids are desirable from one point of view, they provide a healthy diet, fats that are solid at room temperature are required in some foods because of their functional properties. Such applications include the production of non-dairy margarines and spreads, and various applications in the preparation and baking. Many non-dairy fats and animals provide the necessary physical properties, but they also contain either medium-chain cholesteric cholesterol fatty acids. An ideal triglyceride for solid acid applications should contain a predominance of very high melting chain fatty acid, stearic acid, and a monounsaturated fatty acid balance with very little polyunsaturated fat. The fractions of solid fats of natural plants typically have a structure of triacylglycerides with saturated fatty acids that occupy positions sn -1 and sn -3 of triglycerides and an unsaturated fatty acid to position sn -2. This composition of total fatty acid and triglyceride structure confers an optimal solid fat crystal structure and a maximum melting point with the minimum content of saturated fatty acid.

The prototype of natural fat for this vegetable fat of high melting temperature is cocoa butter. More than 2 billion pounds of cocoa butter, the most expensive usable edible oil, are produced around the world. The United States imports several hundred million dollars in value annually in cocoa butter. Volatile and high prices, together with the uncertain supply of cocoa butter, have encouraged the development of cocoa butter substitutes. The fatty acid composition of cocoa butter is 26% palmitic acid, 34% stearic, 35% oleic and 3% linoleic. Approximately 72% of the triglycerides in cocoa butter have the structure in which saturated fatty acids occupy positions 1 and 3 and oleic acid occupies position 2. The composition and distribution of the single fatty acid in cocoa butter in The triglyceride molecule confers properties imminently suitable for the preparation of end uses: they are fragile below 27 ° C and depend on their crystalline state, they rigorously fuse at 25-30 ° C or 35-36 ° C. Consequently, they are hard and not greasy at ordinary temperatures and fuse very rigorously in the mouth. They are also extremely resistant to rancidity. For these reasons, the production of corn oil with increased levels of stearic acid, especially in maize lines containing higher than normal levels of palmitic acid, and reduced levels of unsaturated fatty acids, are expected to produce a substitute of butter. cocoa in the corn. Thus, additional value will be provided to oil and food processors as well as the foreign importation of certain tropical oils will be reduced. The human diet can also be improved by reducing the consumption of saturated fat. Many of the saturated fats in the human diet come from meat products. The diets of chickens and pigs often contain animal fat, which is high in saturated fatty acids, as a source of energy. Non-ruminant animals such as these are very susceptible to the alteration of the fatty acid of the tissue through the modification of the diet (MF Miller, et al (1990) J. Anim. Sci. ,, 68: 1624-31) . A large portion of animal feed ratios are made from corn, which typically contains only about 4% oil. By replacing some or all of the supplemental animal fats in a food ration with the oil present in corn varieties high in oil, which contain up to 10% oil, it will be possible to produce meat products that are low in saturated fats . Food trials in which pigs were fed diets high in oleic acid, showed that the amount of oleic acid deposited in the adipose tissue can be reached substantially without adversely influencing the quality of the meat (MF Miller, et al: LC St John et al. (1987) J. Anim. Sci., 64: 1441-47). The degree of saturation of the fatty acids that comprise an oil determines whether it is liquid or solid. In these studies, the diets of animals high in oleic acid, allow meat quality to be acceptable to the meat processing industry, due to the low level of polyunsaturated fatty acids. Only recently, serious efforts have been made to improve the quality of corn oil, through the progeny of plants, especially after mutagenesis and a wide range of fatty acid composition has been discovered in experimental lines. These findings (as well as those with other oil crops) suggest that the fatty acid composition of corn oil can be significantly modified without affecting the agronomic performance of a corn plant. These are serious limitations that use mutagenesis to alter the fatty acid composition. It is unlikely to discover mutations that a) result in a dominant phenotype ("function gained"), b) are in the genes that are essential for the growth of the plant, and c) are in an enzyme that is not in relationship limitation and that is encoded by more than one gene. Even if some of the desired mutations are available in mutant corn lines, their introgression into elite or better lines by progeny techniques will be slow and costly, since the desired oil compositions in corn are more likely to involve several recessive genes. Recent cell biology and molecular techniques offer the potential to overcome some of the limitations of the scope of mutagenesis, including the need for extensive progeny. Some of the technologies particularly employed are seed-specific expression of foreign genes in transgenic plants [see Goldberg et al. (1989) Cell 56: 149-160], and the use of antisense .RNA to inhibit the target genes of plants in a tissue-specific and dominant manner [see van der Krol et al. (1988) Gene 72: 45-50]. Other advances include the transfer of foreign genes in commercial elite varieties or better from commercial oil crops, such as soybeans [Chee et al. (1989) Plant Physiol. 91: 1212-1218; Christou et al. (1989) Proc. Nati Acad. Sci. U.S.A. 86: 7500-7504; Hinchee et al. (1988) Bio / Technology 6: 915-922; EPO publication 0 301 749 A2], turnip seed [De Block et al. (1989) Plant Physiol. 91: 694-701], and sunflower [Everett et al. (1987) Bio / Technology 5: 1201-1204], and the use of genes as restriction fragment length polymorphism markers (RFLP) in a progeny program, which makes the introgression of traits or qualities in elite lines faster and less costly [Tanksley et al. (1989) Bio / Technology 7: 257-264]. However, the application of each of these technologies requires identification and isolation of commercially important genes. WO 91/13972, published on September 19, 1991, discloses desaturases enzymes relevant for the synthesis of fatty acid in plants, especially desaturases delta-9. U.S. Patent No. 5,443,974, published by Hitz et al. on August 22, 1995, describes the preparation and use of nucleic acid fragments encoding the desaturases ACP stearoyl enzymes of soybean seed or its precursor to modify the composition of the oil of the plant. WO 94/11516, published on May 26, 1994, describes genes for delta-12 miscrosomal desaturases and related enzymes from plants. The cloning of a corn cDNA is described (Zea mays), which encodes the microsomal 12-delta fatty acid desaturase from seed. The discussion of such a citation is thereby incorporated herein for reference. The biosynthesis of oil in plants has been thoroughly studied [see Harwood (1989) in Cri ti cal Revi ews in Plant Sci enses Vol. 8 (l): 1-43]. The biosynthesis of palmitic, stearic and oleic acids occurs in plastids through the reciprocal action of three key enzymes of the "ACP trace"; palmitoyl elongase-ACP, stearoyl desaturase-ACP and acyl thioesterase-ACP. The stearoyl-ACP desaturase introduces the first double bond in the stearoyl-ACP to form the oleoyl-ACP. It is essential in determining the degree of unsaturation in vegetable oils. Due to its key position in the biosynthesis of fatty acid, it is expected as an important regulatory stage. While the natural substrate of the enzyme is stearoyl-ACP, it is shown that I could, like its counterpart in yeast and mammalian cells, desaturate stearoyl-CoA, although scarcely [McKeon et al. (1982) J. Biol. Chem. 257: 12141-12147]. The fatty acids synthesized in the plastid are exported as acyl-CoA in the cytoplasm. At the end three different glycerol acylating enzymes (glycerol-3-P acetyltransferase, l-acyl-glycerol-3-P acyltransferase and diacylglycerol acyltransferase) incorporate the acyl portions from the cytoplasm into the triglycerides during the biosynthesis of the oil. This acyltransferase shows a strong, but not absolute, preference for the incorporation of the saturated fatty acids to the sn-1 positions, and sn -3 and monosaturated fatty acid to sn -2 of the triglyceride. Thus, the alteration of the fatty acid composition of the acyl group will trigger a corresponding change in the fatty acid composition of the oil due to the effects of the mass action. In addition, there is experimental evidence that, due to this specificity and giving the correct composition of fatty acids, plants can produce suitable oils as substitutes for cocoa butter [Bafor et al. (1990) JAOCS 67: 2 1 1 - 225]. Based on the above discussion, an approach to alter the levels of stearic and oleic acids in vegetable oils is by altering their levels in the cytoplasmic clusters of acyl-CoA used for oil biosynthesis. There are two ways to do this genetically. One of these ways is to alter the biosynthesis of stearic and oleic acids in the plastid by modulating the levels of the desaturases of stearoyl-ACP, in seeds through their overexpression or antisense inhibition of their gene. Another conversion of stearoyl-CoA to oleoyl-CoA in the cytoplasm is through the expression of the stearoyl-ACP desaturase in the cytoplasm. To use sense or antisense inhibition of the stearoyl-ACP desaturase in the seed, it is essential to isolate the gene (s) or cDNA (s) encoding the target enzyme (s) in the seed, since any of these inhibition mechanisms require a high degree of complementarity between the antisense RNA (see Stam et al (1997) Annal s of Bo tany 79: 3-12) and the target gene. Such high levels of sequence complementarity or identity are not expected in the stearoyl-ACP desaturases genes from heterologous species. The purification and nucleotide sequences of mammals mammalian stearoyl-Co desaturases have been published [Thiede et al. (1986) J. Bi ol. Ch em. 262: 13230-13235; Ntambi et al. (1988) "Bi ol. Ch em. 263: 17291-17300 and Kaestner et al. (1989) J. Bi ol. Chem 264: 14755-14761.] However, the enzyme of the plant differs from them in being soluble, in the use of a different electron donor, and in their substrate specificities Purification and nucleotide sequences for animal enzymes do not show how to purify a plant enzyme or isolate a plant gene. of stearoyl-ACP was reported from sunflower seeds [McKeon et al (1982) J. Biol. Chem. 257: 12141-12147] and soybeans (US Patent No. 5, 443, 974). Stearoyl-CoA desaturase protein from rat liver has been expressed in E. coli [Strittmatter et al. (1988) J. Biol. Chem. 263: 2532-2535] but, as mentioned above, their specificity of the substrate and the electron donors are completely different from those of the plant. The cDNAs of the stearoyl-ACP desaturases of the plant have been cloned from numerous species including safflower [Thompson et al. (1991) Proc. Nati Acad. Sci. 88: 2578], castor [Shanklin and Somerville (1991) Proc. Nati Acad. Sci. 88: 2510-2514], and cucumber [Shanklin et al. (1991) Plant Physiol. 97: 467-468]. Kutzon et al. [(1992) Proc. Nati Acad. Sci. 89: 2624-2648] have reported that stearoyl-ACP desaturase from turnip seed when expressed in Bra ssi ca rapa and B. nap in an antisense orientation, can result in an 18: 0 level increase in transgenic seeds. The manipulation of stearate levels has been described (Knutzon, D.S. et al., (1992) Proc. Na ti. Acad. Sci. UA 89 (7): 2624-2628). It is possible to raise the level of stearate in seed oils by insufficient expression of the stearoyl-ACP desaturase, the enzyme responsible for the introduction of the first double bond in 18 carbons of fatty acids in plants. The seeds of either B plants. campes tres tri s and B. napus produced by the antisense expression of a cDNA encoding the stearoyl-ACP B desaturase. Farmers using a specific promoter region of the seed, produce oils high in stearic acid, but also contain high levels of linoleic acid (18: 3) when compared to unmodified plants from the same species. Elevated levels of stearic acid have been obtained in soybean by a similar insufficient expression of the stearoyl-ACP desaturase (Patent American No. 5,443,974) and in canola by over-expression of an acyl thioesterase-ACP (U.S. Patent No. 5,530,186). The mutation progeny have also been produced in soybean lines with high levels of stearic acid in their seed oils (Graef, GL Et al., (1985) JAOCS 62: 773-775; Hammond, EG and W, R , Fehr, (1983) Crop Sci: 23: 192-193).

The polyunsaturated fatty acids contribute to the decrease of the melting point of vegetable oils. In high saturated oils, it is a deterioration that lowers its melting point, and therefore higher levels of undesirable saturated fatty acids are still required to reach a plastic fat at room temperature. Additionally, when used in baking for baking and preparation applications, high levels of polyunsaturates lead to oxidative instability as described above for liquid oils. Thus, for maximum utility, a high saturated fat produced in corn should contain saturated fatty acids, mono-unsaturated fatty acid and as much polyunsaturated fatty acid as possible. The combinations of genes discovered in this invention provide new fatty acid profiles in corn, which suggests this criterion. Other combinations result in a lipid profile in which the oleic acid content is not less than 60% of the total oil content. Many of these combinations also utilize a new corn oleosin promoter, or an intron / exon region from the contracted gene 1, or both an oleosin propotor and an intron / exon region from the contracted 1 gene. Lipid reserves in corn seeds are synthesized and stored mainly in a specialized tissue of the embryo called the escutelo. These reserves of lipids constitute up to 50% of the dry weight of the embryo at maturity of the seed. As with all seeds, the lipid stored in corn seeds is packed in simple organelles called oil bodies. These small spherical organelles consist of a triacylglycerol center surrounded by a single layer of phospholipids embedded with proteins called oleosins (Huang (1985) Modern Meth ods of Pl an t Analysi s 1: 175-214; Stymme and Stobart (1987) The Bi ochemi s try of Pl an ts 10: 175-214; Yatsue and Jacks i (1972) Plan t Physi ol. 49: 937-943; and Gurr (1980) The Bi ochemistry of Plan ts 4: 205-248). At least two kinds of oleosin isoforms have been identified in various plant species (Tzen et al (1990) Plant Physi ol. 4: 1282-1289). These two classes are arbitrarily named as isoforms of high molecular weight (H) and low (L), within particular species. The elements of an isoform of various species are understood to be structurally related based on the demonstrations of the properties and possessions in one-sided carrying identity of significant amino acid sequence, and thus, are clearly distinct from the elements of other isoforms (Hatzopoulos et al. al. (1990) Pl an t Cell 2: 457-467; Lee and Huang (1994) Pl ant Mol. Bi ol. 26 (6): 1981-1987; Murphy et al (1991) Bi ochim. Bi ophys. Ta, 1088: 86-94; Qu and Huang (1990) J. Bi o. Chem. 265: 2238-2243). There are three isoforms of oleosins present in corn seeds. They are found in approximately proportional amounts of 2: 1: 1. These isoforms are called OLE16, OLE17, and 0LE18, which correspond to their apparent molecular weights which vary from approximately 16 kDa to 18 kDa. OLE17 and OLE18 are closely related elements of class H, while OLE16 is an element of class L (Lee and Huang, 1994). The genes encoding the three oleosins have been cloned and sequenced (Qu and Huang (1990) J. Biol.Chem. 265: 2238-2243; and Huang, personal communication). These genes are expressed only in the tissues within the embryo (escutelio and axes e briónicos) and the layer of aleurona during the development of the seed, and are positively regulated by the abcisico acid of the hormone (Vanee and Huang (1988) J. Bi ol. Chem. 263: 1476-1491; Huang (1992) Annu. Rev. Plan t Physi ol. Pl an t Mol. Bi ol. 43: 177-200). Oleosins are highly expressed in the embryo, representing approximately 5-10% of the total scutellum protein or 2-8% of the total seed proteins. Promoters of genes that exhibit a specific pattern or expression model of aleuron and embryo ("embryo / aleurone") such as oleosin genes may be attractive candidates for use in transgenic scopes aimed at the expression of a gene that it encodes an oil-modifying enzyme (Qu and Huang (1990) J. Biol. Chem. 265: 2238-2243; and Huang (1992)) and other enzymes of interest for the specific traits of the embryo, especially in corn. Another potential candidate gene from which a corn embryo / aleurone-specific promoter is isolated is the corn globulin-1 gene (Belanger and Kriz, 1989, Plant Physiol., 91: 636-643). However, to date, there is no report describing the expression, regulation or use of such promoters in either temporary expression tests or stably integrated into transgenic corn plants.

BRIEF DESCRIPTION OF THE INVENTION This invention relates to an isolated nucleic acid fragment comprising a corn oleosin promoter, wherein said promoter can be full length or partial and further, wherein said promoter comprises a corresponding nucleotide sequence. substantially the nucleotide sequence in any of SEQ ID NOS: 19 or 38-49 or said promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SEQ ID NOS: 19 or 38-49. In a second embodiment, this invention relates to an isolated nucleic acid fragment encoding a stearoyl-ACP delta-9 desaturase that corresponds sub-substantially to a nucleotide sequence set forth in any of SEQ ID NOS: 8 and 10 or any subfragment. functionally equivalent thereof. Also included are chimeric genes that compensate for such fragments or subfragments thereof or the reverse complement of such fragment or subfragment which are operably linked to suitable regulatory sequences wherein the expression of the chimeric gene results in an altered corn stearic acid phenotype. In a third modality, this invention relates to an isolated nucleic acid fragment encoding a delta-12 desaturase that substantially corresponds to the nucleotide sequence set forth in SEQ ID NO: 2, and any functionally equivalent subfragment thereof, as well as chimeric genes comprising such fragments or subfragments or the reverse complement of such fragment or subfragment which are operably linked to the appropriate regulatory sequences, wherein the expression of the chimeric gene results in an altered maize oleic acid phenotype. In a fourth embodiment, this invention also relates to chimeric genes comprising an isolated nucleic acid fragment encoding a corn stearoyl-ACP delta-9 desaturase substantially corresponding to a nucleotide sequence set forth in any of SEQ ID NOS: 8 and 10 or any functionally equivalent subfragment thereof or the inverse complement of such fragment or subfragment and an isolated nucleic acid fragment encoding a delta-12 desaturase or any functionally equivalent fragment or the reverse complement of such fragment or subfragment which they are operably linked and wherein the expression of such combinations results in an altered corn oil phenotype. Any of these chimeric genes can comprise an isolated nucleic acid fragment comprising a corn oleosin promoter wherein said promoter can be full length or partial and further wherein said promoter comprises a nucleotide sequence substantially corresponding to the sequence of nucleotide in any of SEQ ID NOS: 19 or 38-49 or said promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SEQ ID NOS: 19 or 38-49 or a intron 1 / exon 1 contracted, or both. Also included in this invention are the corn plants and plant parts thereof which contain the various chimeric genes, seeds of such plants, oils obtained from the grain of such plants, animal feed derivatives derived from the processing of such grain, use of the aforementioned oil in food, animal feed, cooking oil or industrial applications, products made from hydrogenation, fractionation, interesterification or hydrolysis of such oil and methods for improving the quality of fresh meat of an animal .

BRIEF DESCRIPTION OF THE LIST OF SEQUENCE AND FIGURES The invention can be more fully understood from the following detailed description and the Figures and Description of the Sequence, which are part of this application. The sequence descriptions summarize the Sequence Listing attached to it. The Sequence listing contains one-letter codes for the character of the nucleotide sequence and three-letter codes for amino acids as defined in the IUPAC-IUB standards described in Nucl ei c Aci ds Research 13: 303 (1985) and in Bi och emi cal Journal 219 (No. 2); 345-373 (1984), and the symbols and formats used for all amino acid and nucleotide sequence data also comply with the rules governing sequence descriptions of amino acid and / or nucleotide in patent applications such as those set forth in 37 CFR & 1,821- 1,825 and Standard St 25. Of the WIPO. SEQ ID NO: 1 is a 1790 nucleotide sequence obtained from a maize cDNA, which encodes a delta-12 desaturase enzyme (fad2-1). This sequence is also exposed in WO 94/11516. SEQ ID NO: 2 is a sequence of 1733 nucleotides obtained from a maize cDNA, which encodes a delta-12 desaturase enzyme (fad2-1). SEQ ID NO: 3 is the translation product of the nucleotide sequence set forth in SEQ ID NO: 3.

NO: 2. The translation product is a polypeptide of 392 amino acids (translation structure: nucleotides 176-1351). SEQ ID NO: 4 is a sequence of 12,313 nucleotides obtained from corn genomic DNA, which comprises the region upstream of the fad2-2 coding region. SEQ ID NO: 5 is a sequence of 2,907 nucleotides obtained from the maize genomic DNA, which includes the intron fad2-l.

"EC ID NO: 6" is an 18-base oligonucleotide primer used to amplify corn delta-9 desaturase, via PCR. SEQ ID NO: 7 is a 17-base oligonucleotide primer used to amplify delta-9 desaturase via PCR. SEQ ID NO: 8 is the 1714 nucleotide sequence of a corn delta-9 desaturase cDNA such as that contained in plasmid pCD520. SEQ ID NO: 9 is the translation product of the nucleotide sequence set forth in SEQ ID NO: 8. The translation product is a 392 amino acid polypeptide (translation structure: 134-1312 nucleotides). SEQ ID NO: 10 is a sequence of 1709 nucleotides of a second delta-9 corn desaturase DNA, such as that contained in plasmid pBN408. SEQ ID NO: 11 is the translation product of the nucleotide sequence set forth in SEQ ID NO: 10. The translation product is a polypeptide of 392 amino acids (translation structure: 102-1280 nucleotides). SEQ ID NO: 12 is an 18-base oligonucleotide primer used to amplify a portion of maize fad2-l via PCR.

SEQ ID NO: 13 is a 17-base oligonucleotide primer used to amplify a portion of maize fad2-l, via PCR. SEQ ID NOS: 14 and 15 are 21-base oligonucleotide primers used to amplify a portion of the 16 kDa gene of oleosin via PCR. SEQ ID NOs: 16 and 17 are oligonucleotide primers of 22, and 20 bases respectively, used to amplify a portion of the 18 kDa gene of oleosin via PCR. SEQ ID NO: 18 is a 46-base oligonucleotide used as a hybridization probe to identify oleosin genes. SEQ ID NO: 19 is a 1714 nucleotide sequence of a 16 kDa corn oleosin promoter. SEQ ID NO: 20 is a 32-base oligonucleotide primer used to amplify deletion derivatives of the 16 kDa promoter from oleosin via PCR. SEQ ID NO: 21 is a 33-base oligonucleotide primer used to amplify deletion derivatives of the 16 kDa oleosin promoter via PCR.

SEQ ID NO: 22 is an oligonucleotide primer of 33 bases used to amplify deletion derivatives of the 16 kDa promoter of oleosin via PCR. SEQ ID NO: 23 is a 32-base oligonucleotide primer used to amplify deletion derivatives of the 16 kDa promoter from oleosin via PCR. SEQ ID NO: 24 is a 37-base oligonucleotide primer used to amplify deletion derivatives of the 16 kDa promoter of oleosin via PCR. SEQ ID NO: 25 is a 32-base oligonucleotide primer used to amplify deletion derivatives of the 16 kDa promoter of oleosin via PCR. SEQ ID NO: 26 is a 32 base oligonucleotide primer used to amplify deletion derivatives of the 16 kDa promoter of oleosin via PCR. SEQ ID NO: 27 is a 33-base oligonucleotide primer used to amplify deletion derivatives of the 16 kDa promoter of oleosin via PCR.

SEQ ID NO: 28 is a 24-base oligonucleotide primer used to amplify deletion derivatives of the 16 kDa promoter from oleosin via PCR. SEQ ID NO: 29 is an oligonucleotide primer of 19 bases used to amplify deletion derivatives of the 16 kDa promoter of oleosin via PCR. SEQ ID NO: 30 is a 25-base oligonucleotide primer used to amplify intron 1 / exon 1 contracted 1 via PCR. SEQ ID NO: 31 is a 25-base oligonucleotide primer used to amplify intron 1 / exon 1 contracted 1 via PCR. SEQ ID NOs: 32 and 33 are 30-base oligonucleotides used as hybridization probes to identify clones containing the globulin-1 gene. SEQ ID NOs: 34 and 35 are 30-base oligonucleotide primers used to amplify the globulin-1 promoter. SEQ ID NOs: 36 and 37 are oligonucleotide primers of 36 and 39 bases respectively, used to amplify the globulin-1 promoter. SEQ ID NO: 38 is a 1.1 kb deletion derivative of the 16 kDa oleosin promoter.

SEQ ID NO: 39 is a 0.9 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 40 is a 0.55 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 41 is a 0.95 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 42 is a 1.4 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 43 is a 1.0 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 44 is a 0.75 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 45 is a 0.4 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 46 is a 1.3 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 47 is a 0.8 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 48 is a 0.6 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NO: 49 is a 0.3 kb deletion derivative of the 16 kDa oleosin promoter. SEQ ID NOs: 50 and 51 are 29-base oligonucleotide primers used to amplify the region encoding fad2-l via PCR.

SEQ ID NOs: 52 and 53 are oligonucleotide primers of 31 and 30 bases respectively, used to amplify the coding region of delta-9 desaturase via PCR. SEQ ID NO: 54 and 55 are oligonucleotide primers of 20 and 25 bases respectively, used to amplify portions of the fad2 genes via PCR. SEQ ID NO: 56 and 57 are 20-base oligonucleotide primers used to amplify the fad2-l intron via PCR. SEQ ID NO: 58 is the complete nucleotide sequence of plasmid pBN257. It contains a start of translation of structure output for fad2-l starting at position 1978. SEQ ID NO: 59 is a truncated form of the fad2-l gene from pBN257. The coding structure of pBN257 is represented by nucleotides 1991-3136 of SEQ ID NO: 58. Figure 1 shows the spotting analyzes Northern regulation of the development of genes that are highly expressed in the embryo and aleurone. The individual stains used the following as probes: Figure 1A, fad2-l; Figure IB, delta-9 desaturase; Figure 1C and Figure ID, • globulin-1, and Figure IE and 1F, 16 kDa oleosin. Figure 2A shows a restriction map of the plasmid pML63. Figure 2B shows a restriction map of the plasmid of pSH12. Figure 2C shows a restriction map of the pSMIOO plasmid. Figure 3A shows a restriction map of plasmid pBN256. Figure 3B shows a restriction map of plasmid pBN257. Figure 3C shows a restriction map of plasmid pBN264. Figure 3D shows a restriction map of plasmid pBN262. Figure 3E shows a restriction map of plasmid pBN414. Figure 3F shows a restriction map of plasmid pBN412. Figure 4A shows the lipid profiles of individual seeds obtained from the corn line FA0132-4.

Figure 4B is a histogram showing the segregation analysis of the lipid profiles of individual seeds obtained from the corn line FA0132-4. Figure 5 shows the lipid profiles of individual R2 seeds obtained from corn line FA013-3-2-15. Figure 6 shows the lipid profiles of individual Rl seeds obtained from corn line FA014-5-1. Figure 7A shows a restriction map of plasmid pBN427. Figure 7B shows a restriction map of plasmid pBN428. Figure 7C shows a restriction map of plasmid pBN431.

DETAILED DESCRIPTION OF THE INVENTION In the context of this description, a number of terms should be used. As used herein, an "isolated nucleic acid fragment" is a RNA or DNA polymer that is single- or double-stranded, optionally containing altered or unnatural, synthetic nucleotide bases. An isolated nucleic acid fragment in the form of a DNA polymer can comprise one or more segments of cDNA, genomic DNA or synthetic DNA. The terms "subfragment that is functionally equivalent" and "functionally equivalent subfragment" are used interchangeably herein. These terms refer to a portion or subsequence of an isolated nucleic acid fragment in which the ability to alter the expression of the gene or to produce a certain phenotype is retained whether or not the fragment or subfragment encodes an active enzyme. For example, the fragment or subfragment can be used in the design of chimeric genes to produce the desired phenotype in a transofmed plant. Chimeric genes can be designed to be used in co-suppression or antisense by binding to a nucleic acid fragment or subfragment thereof, whether or not they encode an active enzyme, in the proper orientation relative to the promoter sequence of the plant. The terms "substantially similar" and "substantially corresponding" as used herein, refer to nucleic acid fragments wherein changes in one or more nucleotide bases do not affect the ability of the nucleic acid fragment to mediate expression of the gene or produce a certain phenotype. The terms also refer to modifications of the nucleic acid fragments of the present invention, such as deletion or insertion of one or more nucleotides that do not substantially alter the functional properties of the resulting nucleic acid fragment relative to the initial unmodified fragment. It is therefore understood, as those skilled in the art will appreciate, that the invention encompasses more than the specific exemplary sequences. However, the skilled artisan recognizes that the substantially similar nucleic acid sequences encompassed by this invention are also defined by their ability to hybridize under moderately stringent conditions (eg, 0.5 x SSC, 0.1% SDS, 60 ° C) with the sequences exemplified herein, or to any portion of the nucleotide sequences reported herein and which are functionally equivalent to the promoter of the invention. Preferred substantially similar nucleic acid sequences encompassed by this invention are those sequences that are 80% identical to the nucleic acid fragments herein or which are 80% identical to any portion of the nucleotide sequences reported herein. The nucleic acid fragments are more preferred, which are 90% identical to the nucleic acid sequences reported herein, or which are 90% identical to any portion of the nucleotide sequences reported herein. More preferred are fragments of nucleic acid, which are 95% identical to the nucleic acid sequences reported herein, or which are 95% identical to any portion of the nucleotide sequences reported herein. Sequence alignments and similarity calculations in percent can be determined using the Megalin program of the LASARGENE bioinformatics computation series (DNSTAR Inc., Madison, Wl). The multiple alignment of the sequences is done using the Clustal alignment method (Higgins and Sharp (1989) CABIOS 5: 151-153) with the default peters (GAP FAULT = 10, GAP LENGTH FAULT = 10). The omission peters for alignments in pairs and the calculation of the percent identity of protein sequences using the Clustal method are KTUPLE = 1, FAULT GAP = 3, WINDOW = 5 AND DIAGONALS CONSERVED? = 5. For nucleic acids these peters are FAULT GAP = 10, FAILURE OF LENGTH GAP = 10, KTUPLE = 2, FAULT GAP = 5, WINDOW = 4 and DIAGONALS CONSERVED = 4. A "substantial portion" of an amino acid or nucleotide sequence comprises sufficient of the amino acid sequence of a polypeptide of the nucleotide sequence of a gene to provide a putative identification of such a polypeptide or gene, either by manual evaluation of the sequence by one skilled in the art, or by computerized sequence comparison and identification using algorithms such as BLAST (Altschul, SF et al., (1993) J. Mol.Bil. 215: 403-410) and Gapped Blast (Altschul , SF et al., (1997) Nucl ei c Acids Res. 25: 3389-3402); see also www.ncbi.nlm.nih.gov/BLAST/). "Gene" refers to a fragment of nucleic acid expressing a specific protein, including the preceding regulatory sequences (5 'non-coding sequences) and then (3' non-coding sequences) of the coding sequence. "Native gene" refers to a gene as it is found in nature with its own regulatory sequences. "Chimeric gene" refers to any gene that is not a native gene, comprising the regulatory and coding sequences that are not found together in nature. Accordingly, a chimeric gene can comprise regulatory sequences and coding sequences that are derived from different sources, or regulatory sequences and coding sequences derived from the same source, but arranged in a manner different from that found in nature. "Endogenous gene" refers to a native gene in its natural location in the genome of an organism. A "foreign" gene refers to a gene that is not normally found in the host organism, but that is introduced into the host organism by gene transfer. Foreign genes can comprise native genes inserted into a non-native organism, or chimeric genes. A "transgene" is a gene that has been introduced into the genome by a transformation procedure. "Coding sequence" refers to a DNA sequence that codes for a specific amino acid sequence. "Regulatory sequences" refer to nucleotide sequences located upstream (5 'non-coding sequences), within, or downstream (3' non-coding sequences) of a coding sequence, which influences transcription, RNA processing or stability, or translation of the associated coding sequence. Regulatory sequences may include, but are not limited to, promoters, leader translation sequences, introns, and polyadenylation recognition sequences. "Promoter" refers to a DNA sequence capable of controlling the expression of a coding sequence or functional RNA. The promoter sequence consists of proximal upstream and more distant elements, the latter elements are often referred to as enhancers. Consequently, an "enhancer" is a DNA sequence which can stimulate the activity of the promoter and can be an innate element of the. promoter or a heterologous element inserted to increase the level or specificity of the tissue of a promoter. The promoters can be derived in their entirety from a native gene, or they can be composed of different elements derived from different promoters found in nature, or still comprise the synthetic DNA segments. It will be understood by those skilled in the art, that different promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental conditions. Promoters that cause a gene to be expressed in most cell types at more times are commonly referred to as "constitutive promoters". The new promoters of various types employed in plant cells are constantly being discovered; Numerous examples can be found in the compilation by Okamuro and Goldberg (1989, Bi ochemi s try of Pl. 15: 1-82). It is further recognized that since in most cases, the exact boundaries of the regulatory sequence have not been fully defined, the DNA fragments of some variation may have identical promoter activity. An "intron" is a sequence that intervenes in a gene that does not code for a portion of the protein sequence. Thus, such sequences are transcribed in the RNA but are then excised or cut and not translated. The term is also used to excise or cut the RNA sequences. An "exon" is a portion of the sequence of a gene that is transcribed and found in the mature messenger RNA derived from the gene, but it is not necessarily a part of the sequence that encodes the product of the final gene. The term "intron / exon contracted 1 or shortened" refers to a region of gene 1 contracted or shortened from corn. The particular intron / exon used in the present invention is derived from a non-coding region ("exon / intron 1") of shortened gene 1 and is identical to the sequence at GenBank access # X02382 of nucleotides 1138 through 2220. As aguí was used, the shortened terms 1 and its abbreviation Sh 1, are used interchangeably. The "leader sequence of translation" refers to a DNA sequence located between the promoter of the sequence of a gene and the coding sequence. The translation leader sequence is present in the processed mRNA completely upstream of the translation initiation sequence. The translation leader sequence may affect the processing of the primary transcript to the mRNA, mRNA stability or translation efficiency. Examples of translation leader sequences have been described (Turner, R. and Foster, G.D. (1995) Molecul ar Bi o technolgy 3: 225). The term "3 'non-coding sequences" refers to DNA sequences located downstream of a coding sequence and includes polyadenylation recognition sequences and other sequences encoding regulatory signals capable of affecting mRNA processing or gene expression The polyadenylation signal is usually characterized by the affectation of the addition of the tracts or extensions of polyadenylic acid to the 3 'end of a mRNA precursor.The use of different 3' non-coding sequences is exemplified by Ingelbrecht et al. (1989). , Pl an t Cell 2: 671-680.) "Transcribed RNA" refers to a product that results from the catalyzed transcription of the RNA polymerase of a DNA sequence When the RNA in a transcript is a complementary copy perfect of a DNA sequence, it refers, meanwhile, to a primary transcript or it can be an RNA sequence derived from post-transcriptional processing l of a primary transcript and reifere as such to a mature RNA. "Messenger RNA" ("mRNA") refers to an RNA that is without introns and that can be translated into a protein by the cell. The "cDNA" refers to a DNA that is complementary to and synthesized from an mRNA standard using the enzyme reverse transcriptase. The cDNA can be double-stranded or double-stranded by the use of a Klenow fragment of DNA polymerase I. RNA "Sense" refers to an RNA transcript that includes an mRNA and thus can be translated into the protein within a cell or in vi tro. "Antisense RNA" refers to an RNA transcript that is complementary to all or part of a target primary transcript or mRNA and that blocks the expression or accumulation of transcripts of a target gene (U.S. Patent No. 5,107,065). The complementarity of an antisense RNA can be with any part of the transcript of the specific gene, ie, the 5 'non-coding sequence, non-coding sequence 3, introns, or the coding sequence. "Functional RNA" refers to an antisense RNA, a ribosome RNA or other RNA that can not be translated but still has an effect on the cellular process. The term "operably linked" refers to the association of nucleic acid sequences in a single nucleic acid fragment such that the function of one is affected by the other. For example, a promoter is operably linked with a coding sequence when it is capable of affecting the expression of such a coding sequence, ie, that the coding sequence is under the transcriptional control of the promoter. The coding sequences can be operably linked to the regulatory sequences in sense or antisense orientation.

The term "expression" as used herein, refers to the production of a functional end product. The expression of overexpression of a gene involves transcription of the gene and translation of the mRNA into a mature protein or precursor. "Antisense inhibition" refers to the production of antisense RNA transcripts capable of suppressing the expression of the target protein. "Overexpression" refers to the production of a gene product in transgenic organisms that exceed production levels in normal or non-transformed organisms. "Co-suppression" refers to the production of sense RNA transcripts capable of suppressing the expression or accumulation of transcripts of endogenous or foreign genes that are substantially similar or identical (U.S. Patent No. 5,231,020). The co-suppression mechanism can be at the DNA level (such as DNA methylation), at the transcriptional level, or at the post-transcriptional level. "Altered expression" refers to the production of the gene product (s) in transgenic organisms in amounts or proportions that differ significantly from the activity in comparable tissue (organ and type of development) from wild-type organisms. "Mature" protein refers to a polypeptide processed post-transcriptionally, that is, one from which any pre or propeptide present in the primary translation product has been removed. "Precursor" protein refers to the primary product of mRNA translation, ie, with the pre and propeptides still present. The pre and propeptides may be but not be limited to intracellular localization signals. A "chloroplast transit peptide" is an amino acid sequence which is translated in conjunction with a protein and directs the protein to chloroplasts and other types of plastids present in the cell in which the protein is made. "Chloroplast transit sequence" refers to a nucleotide sequence that encodes a chloroplast conduction peptide. A "peptide signal" is an amino acid sequence which is translated in conjunction with a protein and directs the protein to the secretory system (Chrispeels, JJ, (1991) Ann. Rev. Pl an t Phy Mol. Bi ol. 42: 21 - 53). If the protein is directed to a vacuole, a target vacuolar signal (supra) can be added additionally, or if it is to the endoplasmic reticulum, an endoplasmic reticulum retention signal (supra) can be added. If the protein is directed to the nucleus, any present peptide signal should be removed and instead include a nuclear localization signal (Raikhel (1992) Pl an tn. 1 00: 1 627-1 632). "Delta-9 desaturase" (alternatively, "stearoyl-ACP desaturase") catalyses the introduction of a double bond between 9 carbon atoms and 10 stearoyl-ACP to form the oleoyl-ACP. Stearoyl-CoA can also be converted to oleoyl-CoA, albeit with reduced efficiency. "Delta-12 desaturase" refers to a fatty acid desaturase that catalyzes the formation of a double bond between carbon positions 6 and 7 (numbered from the methyl end), (i.e., those corresponding to the carbon positions 12 and 13 (numbered carbonyl carbon) of an 18-carbon long-chain acyl chain As used herein, the terms "nucleic acid fragments encoding a corn delta-9 desaturase" and "acid fragment" nucleic acid encoding a corn delta-12 desaturase "refers to nucleic acid fragments that are derived from a desaturase cDNA or a genomic sequence, but which may or may not produce the active enzymes, eg, such fragment it could be a mutant sequence that does not reach a translated product, or the coding structure has been changed that can reach a different polypeptide, but which is functional for the alteration of the level of the enzyme desaturase. after words, such fragment may be used in the construction of a co-suppressor or chimeric antisense gene to alter the level of the enzyme desaturase and thus, alter the lipid profile of a plant transformed with such a chimeric gene. "Transformation" refers to the transfer of a nucleic acid fragment into the genome of a host organism, resulting in genetically stable inheritance. Host organisms that contain the transformed nucleic acid fragments refer to "transgenic" organisms. The preferred method of transforming maize cells is using "gene trip" or accelerated particle transformation technology (Klein K. et al. (1987) Na t ure (London) 327: 10-13; U.S. Patent No. 4,945,050), or a method mediated by Agrobacterium using an appropriate Ti plasmid containing the transgene (Ishida Y. et al., 1996, Nature Biotech, 14: 745-750). The term "transgenic event" refers to an independent transgenic line that is derived from a single callus clone that contains a transgene. The standard recombinant DNA and the molecular cloning techniques used herein are well known in the art and are described more fully in S.ambrook, J., Fritsch, EF and Maniatis, T. Ml ecul ar Cl oning: A Labora t ory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, 1989 (here later "Sambrook"). 'PCR' "Chain Reaction of the Polymerase "is a technique for the synthesis of large amounts of specific DNA segments, consists of a series of repetitive cycles (Perkin Elmer Cetus Instruments, Norwalk, CT) Typically, double-stranded DNA is denatured by heat, the two primers complementary to the 3 'boundaries of the target segment are annealed at low temperature and then extended to an intermediate temperature.A set of these three consecutive stages comprises a cycle.An "expression construct" is a plasmid vector or a subfragment thereof. comprises the immediate chimeric gene.The selection of the plasmid vector is dependent on the method that will be used to transform the host plants.The person skilled in the art is well aware of the genetic elements that could be present in the plasmid vector for the successful transformation of the selected and propagated host cells containing the chimeric gene.The person skilled in the art recognizes that the events of different independent transformation will result in different levels and models of expression (Jones et al., (1985) EMBO J. 4: 2 4 1 1 -2418; De Almeida et al., (1989) Mol. Gen Gene ti cs 21 8: 18 - 8 6), and thus such multiple events could be selected to obtain lines that present the desired level and pattern of expression. Such selection may be accompanied by Southern DNA analysis. Northern analysis of mRNA expression, Western analysis of protein expression, or phenotypic analysis. An "RO" plant is equivalent to a "primary transformant", which is the plant directly regenerated directly from the tissue culture process after transformation by the biolistic method or mediated by Agrobacterium. The seeds harvested from RIO plants were named Rl or RO seeds: 1. The progenies derived from Rl seeds are Rl plants, and the seeds harvested from Rl plants are R2 or Rl seeds: 2. Future generations are named in accordance with this convention. The "grain" is the caryopsis of corn, which consists of a mature embryo and endosperm which are products of double fertilization. The term "grain" or "maize" represents any variety, cultivar or population of Zea mayz L. "Granillo" comprises ripe maize grains produced by commercial growers for or in use of crops or for sale to consumers in both cases for the purposes preferably of growth or reproduction of the species. The "seed" is the grain of mature corn produced for the purposes of propagating the species and for sale to commercial growers. As used herein, the terms seed, grain and granules can be used interchangeably. The "embryo" or also called "germ", is a young sporophytic plant, before the beginning of a period of rapid growth (germination of the seed). The embryo (germ) of corn contains the vast majority of the oil found in the grain. The structure of the embryo in the cereal grain includes the embryonic axis and the escutelio. The "escutelio" is the cotyledon of a strand of a cereal grain embryo, specialized for the absorption of the endosperm. The "aleurone" is a proteineous matter, usually in the form of small granules, which originate in the outermost cell layer of the endosperm of corn or other grains. A "dominant" quality requires an allele to be dominant with respect to an alternative allele if an individual cell or homozygous horganism, for the dominant allele is phenotypic being indistinguishable from the heterozygote. The other, alternative allele is the recessive. "Recessive" describes a gene whose phenotypic expression is masked in the heterozygote by a dominant allele. "Semi-dominant" describes an intermediate phenotype. in a heterozygote. The term "homozygous" describes an existing genetic condition when the identical alleles reside at locis or corresponding sites on the homologous chromosomes. The term "heterozygous" describes an existing genetic condition when different alleles reside in coresponding loci or sites on homologous chromosomes.

As used herein in the description of "oleic acid content", the term "high oleate" refers to a grain or seed having an oleic acid content of not less than about 60% of the total oil content of the seed , by weight when measured at 0% humidity. "Stearic acid content", the term "high stearate" refers to a grain or seed having a stearic acid content of not less than about 20% of the total oil content of the seed, by weight when measured at 0 % moisture. "Saturated fatty acid" is a fatty acid that contains a saturated alkyl chain. The term "high saturated" refers to a grain or seed having a total saturated fatty acid content of not less than about 30% of the total oil content of the seed, by weight when measured at 0% moisture. Major components of the saturated fatty acid fraction of a grain or seed include but are not limited to palmitic (16: 0), stearic (18: 0), and arachidic (20: 0) acids. An "improved quantity of fresh meat quality" is that amount necessary to improve the quality of fresh meat of an animal The present invention relates to the alteration of lipid profiles in corn In one aspect this invention relates to a fragment of isolated nucleic acid comprising a promoter of maize oleosin, if said promoter can be full length or partial and further, wherein said promoter comprises an nucleotide sequence substantially corresponding to the nucleotide sequence in any of SEQ ID NO. NOs: 19 or 38-49 or said promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SEQ ID NOs: 19 or 38-49 In addition, the fragment or subfragments discussed above can hybridize to the nucleotide sequence set forth in SEQ ID NOs: 19 or 38-49 under moderately stringent conditions. The promoter of the corn oleosin, is able to drive the expression of the gene in a specific manner to the embryo and aleurone at a high expression level. The activity of the strong promoter in the development of maize embryos is best achieved by the use of nucleic acid fragment corresponding substantially to the nucleotide sequence set forth in SEQ ID NO: 39 and an intron element in the expression construct as shown in FIG. discuss in the previous examples. It has been found that the activity of the oleosin promoter is much higher and expressed much earlier in the development of the corn kernel, than in a specific promoter of the maize embryo / aleurone, obtained from the globulin-1 gene. The preferred oleosin promoter has the nucleotide sequence set forth in SEQ ID NO: 39. However, as those skilled in the art will appreciate,. any functional promoter which has the embryo / aleurone specified, is employed in the present invention. Other suitable promoters are well known to those skilled in the art, examples of which are discussed in WO 94/11516, the description of which is herewith incorporated herein by reference. In addition, one skilled in the art will be able to use the methods and analyzes described in the Examples below to identify other promoters with the desired embryo / aleurone expression specificity. For example, by using the immediate optimized oleosin promoter as a contol, it is possible to identify other sequences that function in a similar manner, using the molecular and biological histological characterizations of the embryo / aleurone promoter function, such as the expression levels of a GUS reporter function, synchronizing the expression of the gene that is contemporaneous with seed oil formation, and the specificity of the appropriate tissue. In a second embodiment, this invention relates to an isolated nucleic acid fragment that encodes a corn stearoyl-ACP delta-9 desaturase, which substantially corresponds to a nucleotide sequence set forth in any of SEQ ID NOs: 8 or 10 or any functionally equivalent subfragment thereof. Chimeric genes comprising this nucleic acid fragment or subfragment thereof or the reverse complement of such fragment or subfragment operably linked to the appropriate regulatory sequences can be constructed where the expression of the chimeric gene results in a stearic acid phenotype of altered corn Transgenic plants can be made in which a corn delta-9 desaturase enzyme is present at higher or lower than normal levels or in cell types or developmental stages in which they are not normally found. This could have the effect of altering the level of desaturases delta-9 in these cells. It may be desirable to remove the expression or transcribed accumulation of a gene encoding delta-9 desaturases in plants for some applications. To encompass this, a chimeric gene designed for the co-suppression of endogenous delta-9 desaturases can be constructed by linking a nucleic acid fragment or subfragment thereof encoding the corn delta-9 desaturases to the sequences promoters of the plant. Alternatively, a chimeric gene designed to express the antisense RNA for all or part of the immediate nucleic acid fragment can be constructed by linking the nucleic acid fragment or subfragment in reverse orientation to the promoter sequences of the plant, i.e. link of the inverse complement of the fragment or subfragment. Either the co-suppression or chimeric antisense genes can be introduced into the plants via the transformation where the expression or accumulation of the transcript of the corresponding endogenous genes is reduced or eliminated (Stam, et al. (1997) Annal s of Bo t any 79: 3-12).

The expression of a quality or trait gene in corn grains can be covered by the construction of a chimeric gene in which the coding region of the quality or trait gene and another regulatory element (for example, intron) are operably linked to the 16 kDa oleosin promoter. The chimeric gene can comrpender exon 1 / intron 1 shortened or contracted 1 in the 5 'untranslated sequence to either increase the expression of the gene or stabilize the transcripts of the transgene. The sequence of exon I Shl will remain as part of the leader sequences in the mRNA after division occurs. All, or a portion of the coding sequence of the trait or quality gene, is located 3 'in the exonl / intronl sequence.

Shl, and may be in the sense sense or antisense. Such a chimeric gene can also comprise one or more introns to facilitate the expression of the gene. The position of the element or elements may be in the translation leader sequence as described above, or in the coding region of the trait or quality gene. -The intron elements of other genes, such as actin-1, ubiquitin-1, Adh-1, fad2-l, and fad2-2 can also be used in the replacement of the Shl element to have the same effect. Consequently, any intron element from other genes can be used to practice the instant invention. The non-coding sequences that contain the transcription termination signals can also be provided in the chimeric gene. All or a portion of any of the nucleic acid fragments of the instant invention can also be used as a probe for genetically and physically mapping the genes that are part of, and as a marker for, linking traits to these genes. Such information can be used in seed progeny to develop the lines with desired phenotypes. For example, such fragment can be used as a restriction fragment length polymorphism marker (RFLP). Southern blots (Sambrook) of restriction digested genomic plant DNA can be tested with the nucleic acid fragment of the instant invention. The resulting associated patterns can then be subjected to genetic analyzes using computer programs such as MapMarker (Lander et al., (1987) Genomi cs 2: 174-181) in order to construct a genetic map. In addition, the nucleic acid fragment of the instant invention can be used for Southern blots in probes containing genomic DNAs treated with restriction endonucleases from a series of individuals representing origin and progeny of a defined genetic cross. The segregation of the DNA polymorphism is noted and used to calculate the position of the immediate nucleic acid sequence in the genetic map previously obtained using this population (Botstein, D. et al., (1980) Am. J. Hum. Gene £: 32-314-331). The production and use of probes derived from the plant gene for use in genetic mapping is described in R. Bernatsky, R. and Tanksley, S.D. (1986) Plan t Mol. Bi ol. Repórter 4 (1): 31 - A 1. Numerous publications describe the genetic mapping of specific cDNA clones using the methodology summarized above or variations thereof. For example, F2 intercross populations, hybrid cross populations, randomly matched populations, nearby isogenic lines, and other series of individuals can be used for mapping. Such methodologies are well known to those skilled in the art. Nucleic acid probes derived from the immediate nucleic acid sequence can also be used for physical mapping (i.e., placement of sequences on physical maps; see Hoheisel, JD, et al., In: Nonmammal i an Genomi c Analysi s: A Pra cti cal Gui de, Academic press 1996, pp. 319-346, and references cited here). In a third embodiment, this invention relates to an isolated nucleic acid fragment encoding a corn delta-12 desaturase substantially corresponding to the exposed nucleotide sequence., in SEQ ID NO: 2 or any functionally equivalent subfragment thereof. The gene for the microsomal delta-12 fatty acid desaturase described in WO 94/11516, published May 26, 1994, can be used to practice the instant invention. Chimeric genes comprising such a fragment of nucleic acid or subfragment derived therefrom or the reverse complement of such fragment operably linked to the appropriate regulatory sequences can be constructed wherein the expression of the chimeric gene results in an altered corn oleic acid phenotype. As discussed above with respect to an isolated nucleic acid fragment encoding a delta-9 desaturase, it may be desirable to reduce or eliminate the expression or transcript accumulation of a gene encoding a delta-12 desaturase in plants for some applications. To encompass this, a chimeric gene designed for the co-expression of endogenous delta-12 desaturases can be constructed by linking a nucleic acid fragment or subfragment thereof to a plant promoter seuvance. Alternatively, a chimeric gene designed to express an antisense RNA for all or part of a fragment of this nucleic acid can be constructed for binding of the nucleic acid fragment or subfragment in reverse orientation to the promoter sequences of the plants, i.e. the link of the inverse complement of the fragment or subfragment to the promoter sequences of the plant. Either the co-suppression or antisense chimeric genes can be introduced into the plants via the transformation where the expression of the corresponding endogenous genes are reduced or eliminated. The chimeric genes mentioned above may further comprise (1) an isolated nucleic acid fragment encoding a maize oleosin promoter, wherein said promoter may be full length or partial and further, wherein said promoter comprises a corresponding nucleotide sequence. substantially the nucleotide sequence in any of SEQ ID NOs: 19 or 38-49 or said promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SEQ ID NOS: 19 or 38-49 and / or (2) a shortened intron / exon 1. In a further aspect, the chimeric genes can be constructed to encompass a variety of combinations, including but not limited to the following: A) a chimeric gene comprising a isolated nucleic acid fragment encoding a maize stearoyl-ACP delta-9 desaturase, substantially corresponding to a sequence of nucleotide set forth in any of SEQ ID NOs: 8 or 10, or any functionally equivalent subfragment thereof or the reverse complement of this fragment and a nucleic acid fragment encoding a delta-12 desaturase or any substantially equivalent subfragment thereof or the inverse complement of this fragment or subfragment wherein the fragments or subfragments are operably linked and also where the expression of this chimeric gene results in an altered corn oil phenotype. The nucleic acid fragment encoding a corn delta-12 desaturase enzyme, used in the construction of such a chimeric gene, may be the fragment identified in WO 94/11516 or this fragment may correspond substantially to the nucleotide sequence set forth in SEQ ID NO: 2 or any functionally equivalent subfragment thereof. b) The chimeric gene described above in (a) can still further comprise an isolated nucleic acid fragment comprising a corn oleosin promoter, wherein said promoter can be full length or partial and further wherein said promoter comprises a nucleotide sequence that substantially corresponds to the nucleotide sequence in any of SEQ ID NOS: 19 or 38-49 or said promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SECs ID NOs: 19 or 38-49. c) The chimeric gene described in (a) or (b) above may each additionally comprise a shortened intron / exon 1. d) A chimeric gene comprising (1) an isolated nucleic acid fragment comprising an oleosin promoter of corn, wherein said promoter may be full length or partial and further, wherein said promoter comprises a nucleotide sequence substantially corresponding to the nucleotide sequence in any of SEQ ID NOs: 19 or 38-49 or The promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SEQ ID NOs: 19 or 38-49, (2) an isolated nucleic acid fragment encoding a stearoyl- desaturase. ACP delta-9 of corn that substantially corresponds to a nucleotide sequence set forth in any of SEQ ID NOs: 8 or 10 or a functionally equivalent subfragment thereof or the complement inver of the fragment sunfragment, (3) a nucleic acid fragment encoding a corn delta-12 desaturase, or any functionally equivalent subfragment thereof, or the reverse complement of the fragment or subfragment, and (4) a shortened intron / exon. 1 wherein the fragments are operably linked and in addition, wherein the expression of this chimeric gene results in an altered corn phenotype. In another embodiment, the nucleic acid fragment encoding the delta-12 desaturase corresponds substantially to the nucleotide sequence set forth in SEQ ID NO: 2. a) A chimeric gene comprising (1) an isolated nucleic acid fragment comprising a corn oleosin promoter, wherein said promoter can be full length or partial and further wherein said promoter comprises a nucleic acid sequence substantially corresponding to the nucleotide sequence in any of SEQ ID NOs: 19 or 38 -40 or said promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SEQ ID NOs: 19 or 38-49, (2) a fragment of nucleic acid encoding a desaturase delta-12 corresponding substantially to the nucleotide sequence set forth in SEQ ID NO: any functionally equivalent subfragment thereof, or the complement inverse of this fragment or subfragment, or an isolated nucleic acid fragment, which corresponds substantially to the nucleotide sequence set forth in SEQ ID NO: 58 or 59, or any functionally equivalent fragment thereof, or the reverse complement of this fragment or subfragment and a shortened intron / exon 1 wherein said fragments are operably linked and in addition where the expression of this chimeric gene results in an altered corn oil phenotype. In another embodiment, the nucleic acid fragment encoding the delta-12 desaturase corresponds substantially to the nucleotide sequence set forth in SEQ ID NO: 2. This invention also relates to parts of plants and maize plants thereof , which comprise in their genome several chimeric genes. Corn grains obtained from such plants will have altered corn oil phenotypes. For example, a maize grain obtained from a maize plant comprises in its genome, a chimeric gene comprising an isolated nucleic acid fragment encoding a stearoyl-ACP delta-9 desaturase that corresponds substantially to a nucleotide sequence Exposed in any of SEQ ID NOs: 8 or 10 or any functionally equivalent fragment thereof or the reverse complement of this fragment or subfragment operably linked to a suitable regulatory sequence will have a stearic acid content of not less than about 20% of the content of total oil or a total saturase content of not less than about 35% of the oil content total. The preferred regulatory sequence is the oleosin promoter. This same phenotype will be obtained from this chimeric gene which also comprises an isolated nucleic acid fragment encoding a stearoyl-ACP delta-9 desaturase substantially corresponding to a nucelotide sequence set forth in any of SEQ ID NOs: 8 or 10 or any functionally equivalent subfragment thereof or the inverse complement of this fragment or subfragment and / or a shortened intron / exon 1. A grain of corn that comprises in its genome, a chimeric gene comprising an isolated nucleic acid fragment comprising a corn delta-12 desaturase corresponding substantially to the nucleotide sequence set forth in SEQ ID NO: 1, a functionally equivalent subfragment thereof, or the reverse complement of said fragment or subfragment, or an isolated nucleic acid fragment corresponding substantially to the nucleotide sequence set forth in SEQ ID NO: 58 or 59 or a functionally equivalent subfragment thereof or the reverse complement of such fragment or subfragment, a fragment of isolated nucleic acid comprising a corn oleosin promoter: wherein said promoter can be full length or partial and further, wherein said promoter comprises a nucleotide sequence substantially corresponding to the nucleotide sequence in any of SEQ ID NOs: or 38-49, or said promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SEQ ID NOs: 19 or 38-49, and a shortened intron / exon 1 wherein said fragments are operably linked and further, wherein the expression of the chimeric gene results in a phenotype of oleic acid of altered grain, wherein said grain has an oil content in the range from about 6% to about 10% based on the dry matter and in addition where said oil is comprised of not less than about 60% oleic acid based on the total oil content of the seed, and preferably, not less than about 70% oleic acid based on the total oil content of the seed. Such a corn kernel can be obtained by the Top Cross® grain production method cited in the Examples below. In this method, one of the parents or origin comprises the chimeric gene described above and the other origin or parent comprises a phenotype of high oil in the range of about 12% -up to 20% oil by weight or based on dry matter . Alternatively, one of the parents or origin may comprise both transgenes of the invention, for example, a chimeric gene of this invention, and a high oil phenotype, and the other parent or origin is an elite or better hydride line. A corn kernel obtained from a corn plant comprises in its genome, a chimeric gene comprising an isolated nucleic acid fragment encoding a corn delta-12 desaturase that corresponds substantially to the nucleotide sequence set forth in the SEC ID NO: 2 or any functionally equivalent subfragment thereof or the reverse complement of the fragment or subfragment operably linked to the appropriate regulatory sequences will have an oleic acid content of not less than about 60% of the total oil content. The preferred regulatory sequence is the oleosin promoter. This same phenotype will be obtained from its chimeric gene further comprising an isolated nucleic acid fragment encoding a delta-9 maize stearoyl-ACP desaturase substantially corresponding to the nucleotide sequence set forth in any of SEQ ID NO: 8 or 10 or any functionally equivalent subfragment thereof or the reverse complement of the fragment or subfragment and / or a shortened intron / exon 1. With respect to the chimeric genes discussed above in (a) through (c), which comprise the various combinations of the gene, the corn grains obtained from the plants comprising such chimeric genes will have a total saturated content of not less than about 30% of the total oil content and an oleic acid content of not less than about 30% of the total oil content. This invention also relates to seeds obtained from corn plants that contain some of the chimeric genes discussed above, oil obtained from such grain, animal feed derived from the processing of such grain, the use of such oil in food, animal feed, industrial or cooking applications and products made from the hydrogenation, fractionation, interesterification or hydrolysis of such oil, derivatives made during the production of this oil, and methods for the improvement of the quality of fresh meat of animals. The present invention also relates to a method for improving the quality of fresh meat of an animal, which comprises feeding the animal a fresh meat quality by improving the amount of animal feed derived from the processing of the grains / seeds. obtained from any of the corn plants of the present invention. Vegetable oils are often used in high temperature applications. The oxidation of the oil accelerates in the presence of heat. It is important that an oil be able to withstand these conditions for applications such as frying, baking, roasting, etc. In some cases, antioxidants may be needed to improve stability but not all antioxidants resist high temperatures. In addition, in many cases a food manufacturer does not want to use the oils with added antioxidants, if a label with unadulterated ingredients is desired. Therefore, an oil which is stable to oxidation under high temperatures in the absence of any additive or other processing is highly desirable. The overheating of the oils often leads to the thermal polymerization of the oil and the oxidation products result in a construction as varnished, gummy in the equipment used for heating and excessive foaming of the oil. As a result of oxidation, a variety of degradation products are formed depending on the conditions under which the oil is exposed. High temperature stability can be assessed by exposing the oils to high temperature and monitoring the formation of undesirable degradation products. These include both volatile and non-volatile products and may be hydrocarbons, alcohols, aldehydes, ketones, and acids. The non-volatile components can also be classified into polar and polymerized compounds. The polar and polymerized compounds present in a degraded oil can be analyzed directly by reverse phase high performance liquid chromatography as described in Lin, S.S., 1991, Fats and oils oxidation. Introduction to Fats and Oils Technology (Wan, P.J. ed.), Pages 211-232, Am. Oil Chem. Soc. The oil of this invention can be used in a variety of applications. In general, oxidative stability is related to flavor stability. The oil of this invention can be used in the preparation of food. Examples include, but are not limited to, their uses as ingredients, as covers, as salad oils, as roasting oils, and as frying oils. Foods in which the oil can be used include, but are not limited to, biscuit and snack foods, preparation products, suspensions and finishes, sauces and gravies, soups, pastas and mixtures for breads, baking mixes and doughs. Foods incorporating the oil of this invention can better retain the flavor for periods of time due to the improved stability against oxidation imparted by this oil.

The oils of this invention can also be used as a mixing source to make a mixed oil product. For a mixing source, it is suggested that the oil of this invention can be mixed with other vegetable oils to improve the characteristics, such as the fatty acid composition, taste, and oxidative stability of the other oils. The amount of oil of this invention that can be used will depend on the desired properties suggested to achieve the resulting final mixed oil product. Examples of mixed oil products include, but are not limited to, margarines, shortening, frying oils, salad oils, etc. In another aspect, this invention relates to the industrial use of the oil of this invention for industrial applications such as an industrial lubricant for a variety of end uses, such as a hydraulic fluid, etc. The industrial use of vegetable oils as a base fluid for lubricants has been known for many years. However, the interest has intensified due to environmental relationships on the use of petroleum oils in environmentally sensitive areas. Vegetable oils are easily biodegradable, have low toxicity and have good lubricating characteristics. However, high points emptied and rapid oxidation at high temperatures limit their use. Since the oils of this invention are low in polyunsaturates, as discussed herein, and have high oxidative stability and high temperature stability, these characteristics also make the oil of this invention desirable for industrial applications such as an industrial fluid, i.e. , as an industrial lubricant or as a hydraulic fluid. The additives, which can be used to make industrial lubricants and hydraulic fluids, are commercially available. In fact, some additives have been specially formulated for use with high oleic vegetable oils. The additives generally contain antioxidants and materials which retard the formation of foam, deterioration, mold, etc. The oil is obtained from plants by a grinding process. Corn oil is extracted from grains through the use of either a dry or wet milling process. Wet milling is a multi-step process involving the soaking and grinding of grains and the separation of starch, protein, oil and fiber fractions. A review of the wet milling process of corn is given by S.R. Eckhoff in the Proceedings of the Fourth Corn Utilization Conference, June 24-26, 1992, St. Louis, MO printed by the National Corn Growers Association, CIBA-GEIGY Seed Division and the United States Department of Agriculture. Dry milling is a process by which the germ and calyx of the corn kernels are separated from the endosperm by the controlled addition of water to the kernel and subsequent passage through a degerminating mulch and a series of rollers and sieves. The dry milling industry in the United States produces approximately 50 million pounds of crude corn oil per year, and the wet milling industry produces over one billion pounds of crude corn oil (Fitch, B. (1985 ) JAOCS 62 (11): 1524-1531 The resulting oil is called crude oil.The crude oil can be degummed by the hydration of phospholipids and other neutral and polar lipid complexes, which facilitate their separation from the non-hydrating triglyceride fraction The oil can also be refined by the removal of impurities, mainly free fatty acids, pigments and residual gums.The refining is accompanied by the addition of caustic which reacts with the free fatty acid, to form a soup and phosphatide hydrates and proteins in the crude oil The water is used to wash soap residues formed during the refining The derivative of the soap pile can be used directly in food pair to animals or acidulated to recover free fatty acids. The color is removed through the adsorption with a bleaching earth which removes most chlorophyll and carotenoid compounds. The refined oil can be hydrogenated resulting in fats with various textures and melting properties. Winterization (fractionation), can be used to remove stearin from hydrogenated oil through crystallization under carefully controlled cooling conditions. Deodorization, which is mainly steam distillation under vacuum, is the last stage and is designed to remove the compounds which impart odor or taste to the oil. Other valuables derivatives such as tocopherols and sterols can be removed during the deodorization process. The deodorized distillate contained in these by-products can be sold for production of natural vitamin E and other pharmaceutical products of high value. Refined, bleached (hydrogenated, fractionated) oils and fats can be packaged and sold directly or further processed into more specialized products. Hydrogenation is a chemical reaction in which hydrogen is added to the double bonds of unsaturated fatty acid with the aid of a catalyst such as nickel. High oleic acid contains unsaturated oleic acid, linoleic acid, and lesser amount of linoleic acid, and each of these can be hydrogenated. Hydrogenation has two primary effects. First, the oxidative stability of the oil increases as a result of the reduction of the unsaturated fatty acid content. Second, the physical properties of the oil are changed because the fatty acid modifications increase the melting point resulting in a semi-liquid or solid fat at room temperature. There are many variables which affect the hydrogenation reaction, which, in turn, alters the composition of the final product. Operating conditions including pressure, temperature, catalyst type and concentration, agitation and reactor design are almost the most important parameters which can be controlled. The selective hydrogenation conditions can be used to hydrogenate the more unsaturated fatty acids in preference to the less unsaturated ones. Very light or renewed hydrogenation is often used to increase the stability of liquid oils. In addition, hydrogenation converts a liquid oil to a physically solid fat. The degree of hydrogenation depends on the desired embodiment and the melting characteristics designed for the particular final product. Edible fats, used in the manufacture of baked products, solid fats and edible fats used for commercial frying and roasting operations, and bases for the manufacture of margarine are among the myriads of possible fats and oils products achieved through hydrogenation. A more detailed description of hydrogenation and hydrogenation products can be found in Patterson, H.B.W., 1994, Hydrogenation of Fats and Oils: Theory and Practice. The American Oil Chemists' Society.

"Interesterification" refers to the exchange of the fatty acyl portion between an ester and an acid (acidolysis), an ester and an alcohol (alcoholysis) or an ester and ester (transesterification). The interesterification reactions are achieved using enzymatic or chemical processes. The processes of random or directed transesterification rearrange the fatty acids in the triglyceride molecule without changing the composition of the fatty acid. The modified triglyceride structure can result in a fat with altered physical properties. Targeted interesterification reactions using lipases become of increased interest for high value special products as cocoa butter substituents. The products are commercially produced using interesterification reactions including but not limited to edible fats, margarines, cocoa butter substituents and structured lipids containing medium chain fatty acids and polyunsaturated fatty acids. The interesterification is also discussed in Hui, Y.H. (1996, Bailey's Industrial Oil and Fat Products, Volume 4, John Wiley &Sons.) Fatty acids and methyl esters of fatty acid are two of the most important oleochemicals derived from vegetable oils. Fatty acids are used for the production of many products such as soaps, medium chain triglycerides, polyol esters, alkanolamides, etc. Vegetable oils can be hydrolyzed or separated into their corresponding fatty acids and glycerin. The fatty acids produced from various fat separation processes can be used raw or more often, they are purified in individual fatty acids or in fractions by distillation and fractionation. The purified fatty acids and their fractions are converted into a wide variety of oleochemicals, such as dimer and trimer acids, diacids, alcohols, amines, amides, and esters. Fatty acid methyl esters are fatty acids that are increasingly replaced as starting materials for many oleochemicals, such as fatty alcohols, alkanolamines, α-sulfonated methyl esters, diesel oil components, etc. Glycerin is also obtained by splitting triglycerides using separation or hydrolysis of vegetable oils. Additional references in the commercial use of fatty acids and oleochemicals can be found in Erickson, D.R., 1995, Practical Handbook of Soybean Processing and Utilization, The American Oil Chemists' Society, and United Soybean Board; Pryde, E. H., 1979, Fatty Acids, The American Oil Chemists' Society; and Hui, Y.H., 1996, Biley's Industrial Oil and Fat Products, Volume 4, John Wiley & Sons. As discussed above, this invention includes a transgenic corn plant capable of producing grains having an oleic acid content of not less than about 60% of the total oil content. The high oleate trait is dominant. Therefore, the desired phenotype can be obtained if only one of the parental lines in the seed or grain production scheme contains the trait gene. The timeline for commercial production of corn kernel that has high oleic levels, can be greatly accelerated. In addition, the high transgenic saturated trait is dominant. Therefore, the desired phenotype can be obtained if only one of the parental lines in the seed or grain production scheme contains the quality gene. The distribution line for the commercial production of corn that has high oleic levels can be greatly accelerated. The DNA sequence information disclosed in the present invention can be used to isolate cDNAs and genes encoding the delta-9 and delta-12 desaturases from corn grain. Isolation of homologous genes using sequence dependent protocols is well known in the art. Examples of protocol-dependent protocols include, but are not limited to, nucleic acid hybridization methods, and DNA and RNA amplification methods as exemplified by various uses of nucleic acid amplification technologies (e.g., polymerase chain reaction, chain reaction ligase). For example, the genes encoding the desaturases (either cDNAs or genomic DNAs) may be isolated directly by using all or a portion of the nucleic acid sequences present to create the DNA hybridization probes, which could be used to select the libraries using methodology well known to those skilled in the art. Specific oligonucleotide probes, based on the nucleic acid sequences present, can be designed and synthesized by methods known in the art (Sambrook). However, the complete sequences can be used directly to synthesize DNA probes by methods known to those skilled in the art such as random primer DNA labeling, breakage translation, or final labeling techniques, or RNA probes that use high-throughput systems. transcription in vi tro available. In addition, specific primers can be designed and used to amplify some or all of the sequences present. The resulting amplification products can be labeled directly using amplification reactions or amplification reactions after labeling, and used as probes to isolate the full-length cDNA or genomic fragments under appropriate stringency conditions. It is also well known, by persons skilled in the art, that minor alterations (substitutions, additions or deletions) can be created by the use of various in vitro mutagenesis protocols. In this way, any of the nucleic acid fragments of the present invention can be obtained easily.

EXAMPLES The present invention is further defined in the following EXAMPLES, in which, all parts and percentages are by weight and degrees are in Celsius, unless declared otherwise. From the above discussion and these EXAMPLES, a person skilled in the art can assess the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt various uses and conditions.

EXAMPLE 1 Genomic DNA clones and corn fad2-2 cDNA A corn embryo cDNA library was selected using a radioisotopically labeled DNA fragment obtained by PCR and containing the maize gene for the delta-12 desaturase ("fad2-l", WO 94/11516, and set forth in SEC ID NO: l). A second delta-12 desaturase cDNA clone was identified based on its sequence. The second delta-12 desaturase gene was designed in fad2-2.

The full length cDNA sequence is shown in SEQ ID NO: 2. It encodes a polypeptide of 392 amino acids (translation structure: 176-1351 nucleotides). The coding region of corn fad2-2 carries significant sequence identity with fad2-l: they carry 88% identity at the amino acid level, and 92% at the nucleotide level. They also possess 77% identity to the 5 'region - not translated, and 64% to the 3' end. A full length or portion of the coding region of either one of the genes in its antisense or sense approach can be used to suppress both the fad2-l and fad2-2 genes or gene products, because of the significant homology in the coding region between the fad2-ly and fad2-2 genes, and thus produce a high oleate phenotype in transgenic corn. A genomic clone with a 13 kb insert containing the fad2-2 gene was identified using the maize fad2-l cDNA insert as a probe in a selection of the corn grain genomic DNA library (Mol7 line, in the vector? Fix II, Stratagene, La Jolla, CA). The upstream sequence of the coding region is shown in SEQ ID NO: 4, which contains the upstream regulatory element, the 5'-untranslated region, and an intron of 6.7 kb (nucleotide position at 5651-12301) located within the 5 'region -not translated. The intron division site (/ GT / AG /) is retained. The 5'-leader sequence (nucleotide position 5492-5650, and 12302-12313) flanking the intron matings to the sequence of the 5'-untranslated region of the fad2-2 cDNA. The putative TATA box (TAAATA) is at position 5439-5444, which is 47 nucleotides upstream from the first nucleotide of the fad2-2 cDNA clone. The promoter element of this gene can be used to express a gene of interest in transgenic maize plants.

EXAMPLE 2 Intron fad2-l corn Based on the sequence of intron fad2-2 (SEQ ID NO: 4), the primers (SEQ ID NOs: 54 and 55) were designed for PCR amplification of a fad2-2 fragment from corn genomic DNA for used in site mapping or locus fad2-2.

S'-CTGCACTGAAAGrpTGGCA-S 'SEQ ID NO: 54 5'-AGTACAGCGGCCAGGCGGCGTAGCG-3' SEC lD NO: 55 In addition to the expected 0.8 kb fragment that should result from the amplification of the fad2-2 sequence, a second fragment, 1.1 kb in length, was also produced in the same PCR. The 1.1 kb fragment was purified, sequenced and it was determined that this fragment contains a portion of the intron fad2-1. A new series of primers (SEQ ID NOs: 56 and 57) were designed in accordance with the sequences of this partial intron of 1.1 kb, and of the 5'-untranslated region of fad2-l.

, -AAGGGGAGAGAGAGGTGAGG-3 'SECIDNO: 56 5, -TGCATrGAAGGTGGTGGTAA-3' SECIDNO.57 Using the new primer series and the maize genomic DNA as the standard, a PCR product containing the other half of the intron fad2-l was obtained. The fragment was purified and sequenced. A contig containing the complete intron fad2-l was assembled using the sequence that overlaps with the 1.1 kb fragment. The contig is shown in SEQ ID NO: 5. 9U The comparison of the structures of the corn fad2-l and fad2-2 genes revealed that the locations of the introns were conserved. Both introns were located in the 5'-leader region of the precursor RNA. The intron fad2-l is 11 bases upstream of the start codon (ATG), while the intron fad2 is 27 bases upstream of the start codon. The consensus sequences of the intron separation sites (/ GT-AG /) are conserved in both introns. The comparison of the introns fad2-ly and fad2-2 using the BestFit program (Genetics Computer Group, Madison, Wl, using the algorithm of Smith and Waterman (1981) Advances in Applied Mathematics 2: 482-489) showed 81% identity of sequence in the first 0.76 kb (nucleotide positions 3-765 in intron fad2-l [SEQ ID NO: 5]) and nucleotides 5650-670 in intron fad2-2 [as shown in SEQ ID NO: 4] ), and 73% homology near the end of the intron (nucleotide positions 2619-2893 in the intron fad2-l [SEQ ID NO: 5]), and 12006-12320 in the intron fad2-2 [SEQ ID NO: 4] ]). The internal intron sequences were not conserved. Very few introns of studied plants are larger than 2-3 kb (Simpson and Filipowickz (1996) Plant Mol. Biol. 32:41). The research also indicates that the usually large size of intron fad2-2 was due to the insertion of a seemingly intact copy (approximately 4.8 kb) of a retrotransparable element, Milt (San Miguel et al. (1996) Science 274: 765- 768). This retroelement is inserted in an orientation opposite to the direction of transcription of the fad2-2 gene. The intron fad2-l does not contain this element.

EXAMPLE 3 Cloning and sequencing of corn delta-9 desaturase cDNA The degenerate primers were designed in accordance with the conserved regions of the delta-9 desaturase genes from several species, and were used for PCR. These are expounded in SEQ ID NOs: 6 and 7.

'-GAYATGATHACNGARGA .3 'SEQ ID NO: 6 5'-CCRTCRTACAT AGATG-3' SEQ ID NO: 7 65 Two PCR fragments (520 and 500 bp, respectively, were generated when these oligomers were used as primers and DNA from a corn embryo cDNA library, were used as a standard.) These fragments were purified and used as probes to select a corn embryo cDNA library Two independent clones were isolated (pCD520 and pCD500) These two clones were sequenced and hybridized in crosses between themselves and with soybean delta-9 desaturase gene It was confirmed that only the pCD520 insert was homologous to the soy bean desaturase-9 gene The cDNA sequence is shown in SEQ ID NO: 8. The nucleotide number 1-133 is the 5 'leader sequence. -not translated The coding sequence starts from 134 (ATG), and the stop codon (TAA) is at 1309 and 1312, encoding a polypeptide of 392 amino acids set forth in SEQ ID NO: 9. There are 396 nucleotides in regions 3 '-not translated including l to poly (A) label starting at the position of ñucledtido 1661. There is no obvious polyadenylation signal in this region with the possible exception of an AT rich region (1621-1630), located 31 bases upstream of the poly label ( TO) .

The sequence of the cDNA inserted in pCD520 (SEQ ID NO: 8) was used as an interrogation in a search of a DuPont EST database using BLAST programs and algorithms as research tools (Altschul SF, et al. (1990) J Mol. Biol., 215: 403-410 and Altschul, SF et al. (1997) Nucl ei c Aci ds Res. 25: 3389-3402). An EST was identified by this method, and the complete sequence of the ANDc clone from which it was derived is given in SEQ ID NO: 10. The 5'-untranslated leader sequence is in the nucleotide at the 1- position 101, the coding sequence starts from position 102, and ends with the stop codon (TAA) at position 1278-1280. This sequence also encodes a 392 amino acid polypeptide of the sequence which is listed in SEQ ID NO: 11. The coding region of this second delta-9 corn desaturase gene carries significant homology with that listed in SEQ ID NO. : 8. The sequence carries 63% identity and 83% similarity at the nucleotide level, and 77% identity at the amino acid level. There are 429 nucleotides in the 3'-untranslated region of SEQ ID NO: 10, including the poly (A) tag starting at nucleotide 1626. A signal of i Putative polyadenylation (AATAA) is located at nucleotides 1588-1594.

EXAMPLE 4 Spatial and Destructive Regulation of Desaturases Delta-9 and Delta-12 Northern blot analyzes were performed to investigate the spatial and developmental regulation of the genes involved in lipid biosynthesis in corn embryos. Total RNA fractions were purified from the leaves, pods, spikelets, roots and immature embryos dissected from the developing grains at 15, 20, 25 and 30 days after pollination (DAP): The stained RNA was prepared and hybridized individually with 32P-labeled probes of corn fad2-l, (SEQ ID NO: 1), delta-9 desaturase (SEQ ID NO: 8), 16 kDa oleosin (Vanee and Huang 1987), and globulin 1 (Belanger and Kriz, 1989, Plant Physiol. 91: 636-643). The probes were prepared using specific fragments of the gene, purified as described below.

Using the sequence of fad2-l (SEQ ID NO: 19, the primers (SEQ ID NOs: 12 and 13) were designed to ibridize the 3 'end and were used in PCR with the fad2-l cDNA as the standard.

'-AGGACGCTACCGTAGGAA-3 'SEQ ID NO: 12 5, -GCGATGGCACTGCAGTA-3' SEQ ID NO: 13 An expected 0.16 kb PCR fragment was gel purified, and used as the fad2-l specific probe. A cDNA clone containing the delta-9 desaturase (SEQ ID NO: 8), was digested with EcoRI and Xhol, and a 1.7 kb fragment containing the complete cDNA insert was purified as the probe of the delta-desaturase gene. 9. The specific 16 kDa oleosin probe was a 0.25 kb fragment purified from a PCR, using the corn embryo cDNA library as the standard and primers (SEQ ID NO: 14 and 15) by hybridizing to region 3 '-not translated from the 16 kDa oleosin gene.

'-CTrGAGAGAAGAACCACACTC-3 * SEC IDNO: 14 5'-CTAGACATATCGAGCATGCTG-3 'SECIDNO: 15 A corn genomic clone containing the globulin 1 gene was digested by Xho I and Pst I. A 0.77 kb fragment containing the exon portion 4 / intron 5 / a of exon 5 was purified as the globulin-specific probe 1. The analyzes of the Northern blots are summarized in Figure 1. Both biosynthetic lipid genes (desaturases delta-9 and delta-12) are expressed in all tissues / organs examined through several levels. The expression of the desaturates considered coordinamenter regulated in the hembriones, but have different levels of expression spatially. The transcript homologous to the cDNA fad2-l was more abundant in the embryos to the DAP 15, and the level of message declined towards maturation. The same developmental expression profile was detected for the delta-9 desaturase gene. There are high levels of fad2-l expression in both leaves and spikelets, less in root and low but detectable in the pods. The delta-9 desaturase gene expressed at a lower level was examined in these four examined tissues. To down-regulate the genes encoding the delta-9 desaturase, or the microsomal delta-12 desaturase, a seed-specific promoter which is expressed earlier than the target genes, or at least times that match those of the target gene, It could be highly desirable. Specifically, a promoter that is specific to the embryo / aleurone is desired, since these are the tissues that store oil. The same promoter will be equally suitable for overexpression of a trait gene in the development of maize embryos. Therefore there are two known maize genes which are good sources of promoter sequences, gobulin-1 (Belanger and Jriz, 1989, Plant Physiol. 91: 636-643) and 16 kDa oleosin (Vence and Huang, 1987, J Bipl Chem, 262: 11275-11279). The expression profiles of these genes were also characterized by Northern blot analysis. The stable state level of the globulin-1 transcripts begin to accumulate at 20 DAP and reach a maximum level at a relatively late stage of development (30 DAP). Although the 16 kDa gene of oleosin and globulin-1 are both closely spatially regulated and are expressed only in seeds (Belanger and Kriz, 1989, Plant Physiol., 91: 636-643; Vanee and Huang, 1988, J. Biol. Chem. 163; 1476-1481), the level of expression of the oleosin of 16 kDa is much higher judged by the strong hybridization signal in the embryo samples at all stages of development (15-30 DAP) that were examined. The distribution of the oleosin expression of 16 kDa is both earlier than the globulin-1 gene. Immunofluorescent microscopy showed that the 16 kDa oleosin protein is confined to the embryo and aleurone layer of the developing seeds (Vanee and Huang, 1988, J. Biol. Chem. 163; 1476-1481). Therefore, it is concluded that the 16 kDa oleosin promoter could be superior to the globulin-1 promoter to drive the overexpression of quality genes in maize embryos, and the distribution of expression could be optimal to down-regulate the genes involved in the biosynthetic path.

EXAMPLE 5 Isolation and Sequencing of a Maize Embryo and Aleurone-specific Promoter The expression profile of the 16 kDa oleosin gene was compared to the biosynthetic genes of lipid and globulin-1, such as shown in Figure 1. It was concluded that the 16 kDa oleosin is a very good source from which a specific embryo / aleurone promoter sequence is isolated. Corn oleosin proteins contain three major structural domains: a broadly hydrophilic domain at the N-terminus, an a-helical domain of hydrophobic hairpin at the center, and an a-helical amphipathic domain at the C-terminus. However, the nucleic acid and amino acid sequences of the oleosin 18 kDa and 16 kDa are highly similar only in the central domain (Qu and Huang, 1990, J. Biol. Chem. 265: 2238-2243). The primers (SEQ ID NOs: 16 and 17) were designed based on the published sequence of the 18 kDa oleosin (access # J05212, GenBank).

S'-AGGCGCTGACGGTGGCGACGCT-SEQ ID NO: 16 5'-GTGTTGGCGAGGCACGTGAG-3 'SEQ ID NO: 17 These primers hybridize to the central domain region of the 18 kDa oleosin cDNA sequence. PCR-RT was performed (Perkin-Elmer, Norwalk, CT), using the purified total RNA from the development of maize embryos and the previous primer pairs to generate a single 0.23 kb fragment. The fragment was gel purified, and 32-P-labeled as a probe to select a corn library (Missouri line 17, in the vector? FixII, Stratagene). The positive genomic clones were identified and recovered after three rounds of purification. A specific oligonucleotide of the 16 kDa oleosin ("3221-ATG" SEQ ID NO: 18) was synthesized.

'-ACCTCCCGTCGCACCCCGGTGGTGATCAGCCATGGTAGGCTAGCAG-3 'SEQ ID NO: 18 This oligonucleotide contains a sequence complementary to the flanking sequence of the translation initiation codon of the 16 kDa oleosin gene. Specifically, the oligonucleotide is complementary to the starting region of 12 nucelotides prior to the ATG starter translations and which extend another 33 nucleotides in the coding region). This oligomer was labeled with 32, using [T-32P] ATP and T4 polynucleotide kinase (Life Technologies, Gatihersburg, MD), and was used to select the positive genomic clones described above. One of the clones,? 3221, containing a 15 kb insert, was identified as a strong hybridizing to the oligomer probe. The DNA was purified from clone? 3221, digested with various restriction enzymes, electrophoresed on an agarose gel, and stained on a Zeta-probe nylon membrane (Stratagene). The same labeled 32P oligomer (3221-ATG) was used as a probe by DNA staining restricted? 3221 to identify fragments containing the upstream sequences. Based on the hybridization signal patterns of various restriction digests, and the 16 kDa oleosin cDNA sequence, the DNA 3221 was subcloned as follows. The Δ 3221 DNA was digested with Xho I and Xba I, and cloned into the vector pBluescript (pSK (-), Stratagene) previously cut by the same enzymes. The transformants were selected by hybridization to the 3221-ATG 32P-tagged oligomer. The positive clones were isolated. One of the clones (pBN164) was confirmed by sequencing, which contained the elements of the 5'-leader upstream, and the N-terminal part of the coding region of the 16 kDa gene of oleosin. The 1.7 kb sequence of the region upstream of the 16 kDa gene of oleosin in pBN164 is shown in SEQ ID NO: 19. The transcription initiation site (+1) was identified at the position of nucleotide 1609 in base of the first extension data. These are 92 even bases upstream of the initiating codon of the ATG translation. The putative TATA layer (TATAAA) is located at position 1565-1571, 37-43 base pairs upstream of the transcription initiation site. Another box rich in TA was identified at position 1420-1426. These two TA rich boxes are located in the region that is usually rich in GC for the upstream element. The 5 '-not translated sequence is also rich in GC. There is a GC content of 67% from position 1326 to 1700, in contrast to a GC content of only 38% from position 1 to 1325. Southern blot analysis was conducted using the purified genomic DNA of the LH192 corn line (Holdens Foundation Seeds, IA), hybridized with the specific probe of the 16 kDa oleosin. The result indicates that the 16 kDa oleosin of corn is encoded by one or two genes.

EXAMPLE 6 16 kDa Oleosin Promoter Deletion Assay The relative activities of the oleosin promoters of 1 6kDa of corn, and globulin-1, were analyzed using a temporal expression assay. The 35S promoter of the cauliflower mosaic virus was used as a positive control. The temporal expression cassette used β-glucuronidase (GUS) as the reporter gene, fused to the 3 'end of the nopaline synthase (NOS) gene to provide a polyadenylation signal. The putative oleoresin promoter fragment of 1 6kDa contains the full length (1.7 kb, SEQ ID NO: 19) of the fragment upstream of the 16 kDa oleosin gene. The globulin-1 promoter contains a 1.1 kb fragment upstream of the globulin-1 gene. The DNA plasmid was prepared according to the standard procedures (Wizard Miniprep kit, Promega, Madison, Wl), coated in gold particles, and bombarded in embryos of immature maize dissected from the corn cob at 18-19 DAP. Nine embryos were placed in each plate, and 3 plates were bombarded for each tested construct. After the bombardment, the embryos were incubated at 37 ° in a solution of the substrate containing X-Gluc (Jefferson, 1989, Nature 342; 837-838) for 12 hours, and the blue foci that it developed indicating the expression of the GUS gene. counted under the microscope. The result showed that only minimal promoter activity was provided by the full-length upstream fragment of the 16 kDa oleosin gene, indicating that it may be a negative regulatory element present in this region. A number of promoters of the oleosin of 16 kDa of variant length was designed to remove the potential negative regulatory element, and to determine the optimal length with a high activity without losing its tissue specificity. Progressive deletions of the 3 'and 5' end of this upstream sequence were made using PCR, or by restriction digestion. The primers used in the PCR, and the resulting putative promoter fragments, together with the corresponding nucleotide positions in SEQ ID NO: 19, are shown in Table 1. The exon 1 / intron 1 fragment (nucleotide position 1138- 2220 at access # X02382 GenBank) of the shortened-1 maize gene was cloned into the 5 'region-not translated as described below to further optimize the expression cassette.

Table 1. putative promoter fragments of the 16 kDa oleosin gene fragment pramotor primers used in nucleotide position of the 5'-untranslated sequence (size ßr i kb) PCR8 (as in SEC 101) f.6S (1.7) b 1 -1700 native oleosin of 1ßkDa d? Idßr f? 84 (l-7) au: I, d: J 1-1700 Shld £ 222 (1.1) u: A, d: E 512-1619 Sbl £ 220 (0.9) u : B, d: E 749-1619 Shl £ 218 (0.55) u: C, d: E 1075-1619 Shl 1236 (0.4) * > - 1254-1700 native oleoresin of 16kDa 5'-lidßr 054 < 0.95) a: B, d: H 749-1700 native oleoresin of 16kDa 6'-Bder £ 235 (1.4.}. U: D, d: F 99-1501 Shl £ 231. (1.0) u: A, d: F 512-1501 Shl Í232 (0.75) u: B. d; F 749-1501 Shl 2233 (0.4) u: C, d: F 1075-1501 Shl f227 (1.2) u: D. d: G 99-1346 Shl £ 228 (0.8) u: A, d: G 512-1346 Shl £ 229 (0.6) u: B, d: G 749-1346 Shl £ 230 (0.3) u ': C, d; G 1075-1346 Shl PCR was conducted using plasmid DNA pBN164 as the standard, and downstream (u) and upstream (d) primers specified as indicated, except for fl84, in which pBN168 was used as a standard. A restriction enzyme recognition site (underlined) was constructed in most of the primers to facilitate cloning.

A: y-CTTATGTAATAGAAAAGACASS? ICCATATGG-S '(SEQ ID NO: 20) B: 5-GAGGAGTGAGIíáICCTGATTGACTATCTCATTC-3 '(SEQ ID O.21) C: 5-TCTGGACACCCTACCATTGGATCCTCTTCGGAG-3 '< SEQ ID N032) D: 5 * -AGAGTTej2AI CGTGTACAAC ^^ GGTCTCTGG-3, (SEQ ID NO: 23) E: S'-GCCGCTGATGCTC eCTACGACTACGAGTGAGGTAG-S '(SEQ ID NO: 24) F: 5'-ATGCGGGACTCfi4STCGGGGGCAGCGCGACAC-3, (SEQ ID NO: 25) G: ^ -GTGGCGGGGCCGAATCTCGAGTGGGCCGTAGT-3 '(SEQ IDNO: 26) H: S'-GCCACGTGCCAIÜSTAGGCTAGCAGAGCGAGCT-S- (SEQ IDNO: 27) I: 5-AACACACACC ?? TQeATATCACAG-3 '(SEQ IDNO: 28) J: 5 - GG CTGACTTACGGGTGTC-3 * (SEQ ID NO: 29) The fl68 fragment was obtained by cutting the plasmid pBN164 DNA with Xba I and Ncol. The fragment contains the full length upstream region in pBNl64. (A Ncol site is naturally presented at the position of the translation start codon in the 16 kDa oleosin gene). The fragment f36 was presented in pBN236. PBN236 was obtained by cutting pBN168 with Spe I and Xba I, hard-bound by the Klenow enzyme and religated. c. The transcription initiation site (+1) is at the position of nucleotide 1609 in SEQ ID NO: 19. Therefore, the 5'-leader sequence is considered 1609-1700. D. Includes Shl the exon I / intron sequence (position of nucleotide 1 38-2220, access # X02382, GenBank) of the maize shortened gene.

Three intermediate expression constructs, pML63, pSH12, and pSMIOO were prepared. PML63 (Figure 2A) was derived from the commercially available vector pGEM-9Zf (-) (Promega), with an insert containing the 35S promoter, the GUS coding region, and a 3 'Nos region. The plasmid pHS12 contains an exon fragment. 1 / intron 1 (Shl) of the maize shortened-1 gene, inserted between the 35S promoter and the GUS coding region, of pML63. The Shl fragment (nucleotide position of 1139-2230, access # X02382, GenBank) was obtained using a PCR approach. A pair of primers (SEQ ID NOs: 30 and 31) were synthesized. The upstream primer (SEQ ID NO: 30) contains an Xho I (underlined) and the downstream primer (SEQ ID NO: 31) contains a Neo I site (underlined). These sequences were derived from the published sequence of the corn sucrose synthase gene (X02382, GenBank), in which the DNA of a corn library (Missouri line 17, in the? FixII vector, was used in PCR in the caul). Stratagene) as a pattern.

- CTCTCCCGTCCTCGAGAAACCCTCC-T SEQID O: 30 S-CTTGGCAGCCATGGCTCGATG? TT? .. SEQIDNO.3I The resulting 1.1 kb fragment was gel purified, digested with the enzymes Xho I and Neo I, and inserted into the Xho I and Neo I site of pML63 to start pSH12 (Figure 2B). Plasmid pSMIOO contains a globulin-1 promoter, Sh in the 5'-non-tuaducide region, the GUS gene and a 3 'NOS end (Figure 2C). The globulin-1 promoter was obtained from a genomic clone isolated from a corn library (constructed in EMBL-3, Clontech, Palo Alto, CA), using oligomers labeled at the end (SEQ ID NOs: 32 and 33) as probes in the selection. Oligomer sequences are based on the globulin-1 cDNA sequence available as GenBank access M24845).

-ATGGTGAGCGCCAGAATCGTTGTCCTCCTC-3 'SEQ IDNO.32 5-CATCCTGGCGGTCACCATCCTCAGGAGCGT-3' SEQ ID NO3 - 3 A positive clone with an insert of approximately 10 kb that hybridized to both probes of oligomers was confirmed to have the globulin-1 gene. A 0.45 kb fragment of the 5 'primer codon was obtained from PCR using the 10 kb clone as the standard. The primers used in the amplification of the 0.45 kb segment are presented in SEQ ID NOs: 34 and 35. The upstream coil (SEQ ID NO: 34) contains a site for the EcoRI enzyme (underlined) and the downstream primer contains a site for the Ncol enzyme (underlined).

S'-ATAGGGAATTCTCTGTTTTTCTAAAAAAAA > 3 'SEQIDNO: 34 S'-GCTCACCATGGTGTAGTGTCTGTCACTGTG' SEQ ID NO.35 The fragment was purified and cut with EcORI and Ncol, inserted into a vector with comparable sites for cloning, A fragment of 0.66 kb Hind III-EcoRI immediately upstream of the region of 0. 45 kb was cut from the 10 kb clone and ligated upstream of the O.45 kb fragment, reaching a final 1.1 kb globulin-1 'promoter fragment.

This clone was used in the PCR with the specific primers of the globulin-1 promoter (SEQ ID.

NO: 36 and 37). The upstream primer (SEQ ID NO: 36) contains a site for BamHl (underlined), and the downstream primer (SEQ ID NO: 37) contains a site for Xhol (underlined).

'-GGGGGATCCAAGCTTGAGGAGACAGGAGATAAAA? T-l 'SEQ ID NO: 36 SvGGGCTGCAGCTCGAGGGTGTAGTGTCTGTCACTGTGATA-' SEQ ID NO: 37 The 1.1 kb PCR fragment was purified, digested with BamHl and Xhol, and inserted into the sites BamHl and Xhol of pSH12 to replace the promoter 35S. The resulting plasmid was designed as pSMIOO (Figure 2C). All the 16 kDa promoter fragments of the putative oleosin (listed in Table 1) were gel purified prior to cloning into the expression vector. The fl68 fragment was inserted into the Xbal and Ncol site of pML63 (to replace the original 35S promoter in the construct), and the new construct was called pBN168. The fragments purified by PCR described in Table 1, were digested with the corresponding restriction enzymes designed in the primers (Ba Hl and Xhol for f222, f220, f218, f235, f231, f232, f233, f227, f228, f229 and f230 ), and were inserted into the expression vector (pSMIOO), previously digested by the same enzymes to replace the globulin-1 promoter. The fl84 fragment was cut with Neo I, and inserted into the Ncol site of pBN168. The resulting construct, pBN184, contains the 5'-liner sequence of the native 16 kDa oleosin, with the Sh 1 element in the 5'-untranslated region. F254 fragment was digested with BamHI and Ncol, and inserted into the BamHI / NcoI site of pML63. The different promoters and the 5'-non-translated fragments contained in these constructs are listed in Tables 1 and 2. The sequences of each of these promoters (as derived from the 1.7 kb promoter of lontigud compoleta, and not including the sites of restriction inscribed during cloning) are listed in the sequence listing as follows. SEQ ID NO: 38 is the 1.1 kb promoter fragment, SEQ ID NO: 39 is the 0.9 kb promoter fragment, SEQ ID NO: 40 is the 0.55 kb promoter fragment, SEQ ID NO: 41 is the 0.95 promoter fragment. kb, SEQ ID NO: 42 is the 1.4 kb promoter fragment, SEQ ID NO: 43 is the 1.0 kb promoter fragment, SEQ ID NO: 44 is the 0.75 kb promoter fragment, SEQ ID NO: 45 is the promoter fragment 0.44 kb, SEQ ID NO: 46 is the 1.3 kb promoter fragment, SEQ ID NO: 47 is the 0.8 kb promoter fragment, SEQ ID NO: 48 is the 0.6 kb promoter fragment, SEQ ID NO: 38 is the promoter fragment of 1.1 kb, and SEQ ID NO: 49 is the 0.3 kb promoter fragment. The plasmid DNAs purified from these constructs was used in the temporal expression assays as previously described. The resulting GUS staining tests, indicating the activities of the prmotor, are summarized in Table 2.

Table 2. Suppression Assay of the 16kDa Oleosin Promoter Plastid Builder Promoter Activity to pBN! 68 pOle-I.7kb5 ': GUS :: Nos3' +/- pBN184 pOle-1.7kb5 ':: Sh :: GUS :: Nos3 * - pBN222 pOle-1.1 kb5":: Sh :: GUS :: Nos3 '-H- pBN220 POle-0.9kb5' :: Sh :: GUS :: Nos3 '+ ++ - + H- + pBN2I8 pOle -0.55kb5 ':: Sh :: GUS :: Nos3- • m IM! PBN254 pOIe-0.95kb5 • :: GUS :: os3 • ++ pBN236 pOle-0.4kb5' :: GUS :: Nos3 '++ // - pBN235 pOle-1.4kb5":: Sh :: GUS :: Nos3 'pBN231 pOIc-1, 0kb5" :: Sh :: G? S :: Nos3' pB 232 pOIe-0.75kb5":: Sh :: GUS :: Nos3 'pBN233 pOle-0.4kbS ":: Sh :: GUS :: Nos3' pBN227 pOle-13kb5" ':: Sh :: GUS :: Nos3' + pBN228 pOle-0.8kb5 ~ :: Sh :: GUS: : Nos3 '+ pB 230 pOlc-0.3kb5l ":: Sh :: GUS :: Nos3l + pSMlOO pG¡oI.lkb5' :: Sh :: GUS :: Nos3 '++ to. The activity of the promoter was measured by a temporary expression assay of the GUS reporter gene. The + was assigned based on the visual estimation of the intensity and counts of the blue fuci -: 0, +/-: 0-1; +: 2-10; ++: 10-50; +++ _ 50-100; ++++: 50-100, but significantly more obscure than +++; +++++: > 150 blue fuci The full-length promoter (contained in pBN168 and pBN184), together or not with the intron element SH1, confers minimal or no detectable promoter activity in the temporal expression system. Promoter activity increased when this region was progressively suppressed from the farther upstream end. There seems to be a negative regulatory element in this farther upstream region (1-511). The deletion of this region as in pBN222 significantly increased the GUS expression compared to the activity of pBN184 in the assay. The removal of even more sequence, up to the position of nucleotide 748, in addition, increased the activity of the promoter, as demonstrated with the pBN220 construct. However, promoter activity decreases if the upstream sequence is deleted beyond position 748 (pBN218 versus pBN220). The inclusion of the TATA box (1566-1571) is important to achieve the high promoter activity. However, the TATA-rich element upstream (1420-1436) can be replaced by the TATA box (1566-1571), albeit with significantly less activity. The function of the GC-rich region (1326-1700) surrounding the TAT boxes is not apparent from this data. The minimum promoter activities are still detected when the full GC rich region, including the TATA boxes, are suppressed. The increased intron is very important in the optimization of gene expression. None of the constructs lacking the Shl element provides any significant level of GUS expression in the assay. The 16 kDa oleosin promoter with optimized length and composition, as in pBN220, was found to be stronger than the globulin-1 promoter (contained in pSMIOO). The results of the Northern blot analysis early characterize the distribution of expression in the young development of maize embryos, combined with the demonstration of their high activity in the expression assay, indicating that the optimal embryo / aleurone specific promoter is the 0.9 kb fragment (SEQ ID NO: 39) isolated from the 16 kDa oleosin gene combined with the exon 1 / intron 1 element of Shl (SEQ ID NO: 39), isolated from the combined 16 kDa oleosin gene with an exon 1 / intron 1 element of Shl in the 5'-untranslated region.

EXAMPLE 7 Embryo / Maize Aleurone-specific Expression Constructs with Lipid Trait Genes The expression constructs comprising a 16 kDa oleosin promoter from maize (0.9 kb in length, Table 1 and 2, and SEQ ID NO: 39), an exon 1 / intron 1 (1.1 kb) elongation of the shortened gene. 1 located between (3 'a) promoter and (5' a) the cDNA fragment, a cDNA fragment encoding a portion of the quality gene in either sense or antisense orientation with respect to the promoter and at the 3 'end We located at 3 To the cDNA fragment, they were constructed and used in the transformation of corn to alter the level of the enzyme encoded by the trait gene in corn grains (Figure 3B-3F). The designed construct is suitable to express any target trait gene not mentioned in this patent in a manner specific to the embryo / aleurone. The selectable marker in the vector skeleton can be any antibiotic-resistant gene (eg, ampicillin, hygromycin, kanamycin). An intermediate construct, pBN256, modified from pBN220 was made as the initiator vector for the various expression constructs with the lipid trait genes. PBN220 was digested with Ncol and EcoRI to suppress the GUS coding sequence, the end filled with dNTPs and the Klenow fragment of DNA polymerase I, and religated. The resulting plasmid was designated pBN256 (Figure 3A). PCR was used to obtain a fragment containing the fad2-l coding region with a Kpn I restriction site at both ends. The fad2-l cDNA clone was used as the template with the primers (SEQ ID NOs: 50 and 51) specific to the fad2-l sequence each containing a site for Kpnl (underlined).

, -CGGGJ2 £ CCGATGACCGAGJ \ AGGAGCGGG-3 'SEQ ID OS.50 5 * -GGCGÜTA? £ TAGAACTTCTTGTTGTACCA-3' SEQ ID NOS: 51 The expected 1.2 kb fragment was gel purified, digested with Kpn I, and cloned into a vector with a comparative Kpn I site to facilitate further propagation and manipulation. The Kpnl fragment was digested from this new construct, and the ends were direct as previously, inserted into the Sma I site of pBN256, to become pBN257. This clone contains a complete near length of the coding region fad2-l, but the initiating codon of the ATG translation is outside the structure (Figure 3B). A DNA fragment containing the delta-9 desaturase coding region was recovered by PCR using the DNA clone of delta-9 desaturase cDNA clone (SEQ ID NO: 8) as the template and the specific primers of the coding region (SEQ ID NOs: 52 and 53) containing the Ncol sites.

The resulting fragment was gel purified, cut by Ncol, and inserted into the Ncol site of the modified pBN220 in which the GUS gene has been previously removed.

• -GGCCTCCGCI ^ IG ^ CGCTCCGCTCCACGACG-3, SEQ IDNOS: 52 S'-CTCCAACTCAAGCAGTCGC ^ AIG ^ GTTTCC-S SEQ IDNOS: 53 (Plasmid pBN220 was cut by Ncol and Sma I to remove the GUS gene, and it was fully filled by Klenow treatment, and religated as the modified GUS free vector). The resulting clones contain a truncated delta-9 desaturase coding region (approximately 0.9 kb, comprising 79% of the full-length coding sequence) in each of the two possible orientations, sense (pBN264, FIG. 3C) 'and antisense (pBN262, Figure 3D). The 0.9 kb Neo I fragment of the delta-9 desaturase gene (SEQ ID NO: 8) was also cloned into the Neo I site of pBN257 to create a construct, pBN414 which contains a fused trait gene of the desaturated fad2 -ly delta-9, both in the sense orientation as shown in Figure 3E). The coding sequence of fad2-l in pBN414 is outside the structure as in pBN257, and its C-terminal sequence is interrupted by the insertion of the delta-9 desaturase fragment (79% of the full length coding region shown in FIG. SEC? .D NO: 8). The second delta-9 desaturase clone (SEQ ID NO: 10) was cut by EcoRI, and the 1.1 kb EcoRI fragment was purified and inse into the EcoRI site of pBN257 to create a new construct, pBN412 (Figure 3F), which contains a fused trait gene of delta-9 and fad2 desaturase, both in orientation sense. In pBN412, the fragment of the delta-9 desaturase contains a full-length coding region (SEQ ID NO: 10). The initiating codon of the ATG translation for the delta-9 desaturase is in the pBN412 structure, but the fad2 encoding the sequence is outside the structure.

EXAMPLE 8 Transgenic corn to. Corn Transformation The chimeric genes described above were introduced into corn cells by the following procedure. The immature corn embryos were dissected from the developing cariopses derived from crosses of inbred corn lines H99 and LH132, or crosses from inbred corn lines H99 and LH195, or a public High II line (Armstrong, 1991, Maize Genetics Co. News Letter 65: 92-93), or any line of corn, which are transformable and regenerable. The embryos were isolated 10 to 11 days after pollination when they were 1.0 to 1.5 mm in length. The embryos were then placed with the axial shaft face down and in contact with the solidified N6 agarose medium (Chu et al., (1975) Sci. Sin. Pekin 18: 659-668). The embryos were kept in the dark at 21 °. Friable embryogenic callus proliferated from the scutellum of these immature embryos. They consist of undifferentiated masses of cells with somatic proembryoids and carrying embryoids in the suspensory structures. The embryogenic calli isolated from the primary explant can be cultured in an N6 medium and subcultured in this medium every 2 or 3 weeks. The plasmid, p35S / Ac (obtained from Dr. Peter Eckes, Hoechst Ag, Frankfurt, Germany) can be used in the transformation experiments together with the trait gene (cobombard) to provide a selectable marker. This plasmid contains the gene Pa t (see European Patent Application 0 242 236), which encodes the transferase of acetyl phosphinothricin (PAT). This gene is from S trept omyces viri dochromogenes, and its sequence is located in GenBank access X65195. The PAT enzyme confers resistance to inhibitors of the herbicidal glutamine synthetase, such as phosphinothricin (also available as the compound designated glufosinate). The pa t gene in p35S / Ac is under the control of the 35S promoter of the Cauliflower Mosaic Virus (Odell et al (1985) Na t ure 313: 810-812) and the 3 'region of the nopaline synthetase gene (3 'NOS end) of the T-DNA of the Ti plasmid of Agroba ct eri um turne asci ens. Alternatively, the gene fragment pa t purified in gel, including the 35S promoter, the coding region of the pa t gene and the 3 'end NOS, can be used as the selectable markup. It will be appreciated by those skilled in the art that the fragment used to provide selection in the transformations can vary considerably, and that any fragment containing the 35S promoter operably linked to the pa t gene is capable of providing the desired selectable feature. Another gene that is employed as a selectable marker for resistance to phosphinothricin and which can be provided in a plasmid or as a separate DNA fragment, is the bar gene of S trep t omyces hydroscopius (GenBank access X17220). The particle bombardment method (Klein et al. (1987) Na t ure 327: 70-73) was used to transfer genes to the callus of the culture waxes. In accordance with this method, the gold particles (0.6 μm or 1 μm in diameter) were coated with DNA using the following technique. Approximately 10 μg of the plasmid DNAs were added to 50 μL of a suspension of gold particles (60 mg per mL). Calcium chloride (50 μL of a 2.5 M solution) and free base spermidine (20 μL of a 1.0 M solution) were added to the particles. The suspension was vortexed during the addition of these solutions. After 10 minutes, the tubes were briefly centrifuged (5 sec at 15,000 rpm) and the supernatant was removed. The particles were resuspended in 200 μl of absolute ethanol, centrifuged again and the sobendant was removed. The ethanol rinsing was performed again and the particles were resuspended in a final volume of 30 μL of ethanol. An aliquot (5 μL) of the golden particles coated with DNA was then placed in the center of a Kaptona fast disk (Bio-Rad Labs). The embryogenic tissue was placed on the filter paper on a N6 solidified agarose medium. The tissue was arranged as a thin lining covering a circular area approximately 5 cm in diameter. The petri dish containing the tissue was placed in the PDS-1000 / He chamber approximately 8 cm from the detection selection. The air in the chamber was evacuated to a vacuum of 28 inches Hg. The particles coated with DNA were accelerated in the corn tissue with a Biolistica PDS / 1000 / He (Bio-Rad Instruments Hercules CA), using a helium pressure of 1000 psi, a gap distance of 0.5 cm and a fast distance of 1.0 cm. Seven days after the bombardment, the tissue was transferred to an N6 medium containing glufosinate (5 mg per liter) and lacks casein or proline. The tissue continued to grow slowly in this medium. After an additional 2 weeks, the tissue was transferred to fresh N6 medium containing glufosinate (selection medium). The tissue was cultured in the selection medium and transferred every 2 weeks for a total of 3-4 passes. The approximately 1 cm diameter areas of actively growing calli were identified on some plates containing the selection medium. This callus continued to grow when it was sub-cultivated in the selective medium. The plates were regenerated from the transgenic calluses by the clusters or tissue transfer cells of the N6 medium supplemented with 0.2 mg per liter of 2,4-D (regeneration medium). After 2-3 weeks, the tissues begin to form structures similar to the somatic embryo and show green areas when the tissue area is transferred and grow under the light. The seedlings emerged after a total of 3-4 weeks in the regeneration medium, and were individually transferred into the tissue culture flasks of the plants containing the regeneration medium. After successive growth of the root and stem, the seedlings were transplanted into 4-inch pots in the growth chamber, and after replanting in 10-12-inch pots, they were grown to mature in the greenhouse (Fromm et al. al., (1990) Bi o / Technolgy 8: 833-839). b. Transgenic corn with High Saturated Saturated Grain Acid Composition Using the biolistic firing method described above, the corn callus was co-bombarded with the delplasmid DNA pBN262, and the bar gene fragment. The stable transformants were selected in accordance with the procedures described above, and the transgenic corn plants were selected in accordance with the procedures described above, and the corn plants were regenerated. The primary transformants (designed as RO plants) were grown in the greenhouse. The plants were either crossed or the same using the wild-type pollen of the line LH132 Holdens. The ears of corn were harvested at DAP. The embryos were dissected from the grains, and sterilized. The small pieces of the scutellum were taken from each individual connection and used for the fatty acid composition tests by the GC method as described in WO 04/11516. The remaining embryos were planted in the tissue culture vessels containing the regeneration medium. Embryos with a positive phenotype (ie, a high level of saturated fatty acids in the lipid fraction) were transplanted from the containers of potted crops, and were grown in Rl plants in the greenhouse. Mature Rl plants were either the same or crossed with wild-type pollen (from line 5-12-24, Pioneer Hybrid International, Johnston, IA). The ears of corn were harvested at 45 DAP, and the grains R2 were collected. The small pieces of the scutellum were taken from the individual grains and used for the analysis of their fatty acids. Two independent transgenic lines were identified as having a high saturated fatty acid phenotype, FA013-2-4 and FA013 3-2. Figure 4A shows a typical example of the Rl grain phenotype: 2 segregants of a single ear of corn harvested from an Rl plant of line FA013-2-4. The RO generation of this plant was cross-pollinated with a wild-type pollen from LHÍ32 (Holden). The corn cob was harvested and the lipid composition of the individual grains was analyzed. The results show a segregation of the 1: 1 seed (high saturated phenotype or wild type) indicating the presence of a locus or unique transgenic insertion site in FA013-2-4. A heterozygous grain containing 26-1% stearic acid (against wild type such as 2%) was planted and grown in a Rl plant. The Rl plant was the same, and the analysis data for the R2 seeds indicated a segregation ratio of 3: 1 (Figure 4A and 4B), confirming that the FA013-2-4 contains a locus or single insertion site of the transgen, and that the phenotype of the trait is dominant. In the "segregants of the R2 seeds, the stearate content in the grains varied from 27-43%, and the average fatty acid composition was 13% 16: 0, 37% 18: 0, 4% 18: 1, 39% 18: 2, 2.8% 18: 3, and 0.5% 20: 0 and 20: 1. The saturated total fatty acid content was 54% The maximum saturated fatty acid content was found as high as 61%. This was in a line that had a total composition of 13% 16: 0, 43% 18: 0, 3% 18: 1, 34% 18: 2, 2.3% 18: 3, 4.6% 20: 0, and 0.2% 20: 1. This was compared with the composition of the segregating profile of the native type of 16% 16: 0, 2% 18: 0, 19% 18: 1, 63% 18: 2, 1.0% 18: 3, and 0.1% 20: 0. Native-type segregants have a total saturated fatty acid content of 18% The germination rate of the seed of the FAP13-2-4 line is close to 100% in the growth chamber conditions standard, indicating that the content of saturated fatty acid in the embryo / aleurone does not affect the viability of the seed. a typical example of the phenotype and segregation of Rl grains: 2 harvested from two Rl plants of line FA013-3-2-15. Their respective RO plants were the same, and the corresponding Rl plants were both pollinated in cross with wild type pollen from line 5-12-24. The first plant was derived from an RO: 1 grain that originally contains 12% stearate, and the second plant of a grain with 21% stearate content. However, the maximum stearate content of the Rl: 2 grains of both plants reached up to 38-39%. The variation range of stearate levels in the Rl: 2 grains was 29-38%, and 16-39% respectively. This indicates the presence of a locus or single transgene insertion site in line FA013-3-2-15 based on the proportion of segregation. The average total saturated content was more than 50%, and the germination rate of the seed for this line was approximately 40%. The seeds R3: 4 were obtained from a homozygous plant of 1 event FA013-2-4. The composition of the homozygous grain lipid was, on average, 15% 16: 0, 15% 18: 0, 14% 18: 1, 53% 18: 2, 1.5% 18: 3, 1.5% 20: 0, and 0.5 % 20: 1. However, the grains harvested from a heterozygous plant with the same generation R3: 4, contains a higher stearate content (31% versus 15% of the homozygous background). A similar result was obtained in the grains harvested from the crosses using a heterozygous plant as the pollen donor in a hybrid female plant (34K77, DuPont), in the grain production method. TopCross® (TC) (Table 3).Table 3. Composition of the grain IFpido in RO: 1, R-: 4 homozygotes and histaterozygotes, and several crosses of FAOI 3-2-4 Genotype Phenotype (%) 16: 0 18: 0 18: 1 18: 2 18: 3 20: 0 20: 1 R0: I x LH132 14 23 12 47 R3: 4 the same (homozygous) 15 15 14 53 1.5 1.5 0.5 R3: 4 the same (heterozygote) "12 31 8 44 2.4 3.0 0.3 34K77 (TC) R3 * 12 32 7 45 2.1 2.7 0J T * 15 1.2 18 65 0.7 0.3 0J the data represent the composition of * average lipid of the grains with the positive phenotype. The grains of R3: 4 were from the same corn ears of the heterozygous R3 plant. The same R3 plant was used as the pollen donor to pollinate 34K77 plants. A few 34K7 plants were the same to obtain the native type grains as the control.

Using procedures similar to those described above, new transgenic events with high phenotypes were generated in stearate - and here high saturated - (Table 4). The quality gene constructs used in these experiments were either pBN264 or pB247 (Figure 7A). Plasmid pBN264 is similar to pBN262, except that the delta-9 desaturase is in a sense orientation relative to the promoter. The transgenic sequence is contained within a Sal I fragment (position 3248-44) or pBN427 and is identical to the corresponding Sa I fragment of pBN264 (position 2-3206). However, pBN427 is a vector structure with a selectable hygromycin resistance marker (HPT, from pKS17, described in WO 94/11516), against the ampicillin marker in pBN262 and pBN264. The transgene prepared for bombardment was either the digested restriction enzyme and the purified DNA fragment on agarose gel of pBN264 (for events derived from the FA025 experiment, the transgenic fragment was labeled as fBN264), the plasmid DNA pBN427 (for events derived from experiment FA029). The restriction enzyme used to cut the transgene may be Sal I or Xba I, which release the transcriptionally functional 3.2 kb transgene fragment, which can be purified following agarose gel electrophoresis. The use of a transgenic DNA fragment, instead of the total plasmid, allows the recovery of transgenic events, which do not contain a resistance gene to the bacterial antibiotic.

Table 4. Transgenic events with high stearate phenotype Transgenic events Stearate "Total sat.?> Construct native type Co-sup frec d < 2% 18% 1) FA025-1-4 16-27% 32-42% ÍBN264 2) FA025-2-1 12-39% 28-60% ÍBN264 3) FA025-2-12 17-39% 50-55% ÍBN264 6/30 = 20% 4) FA025-2-17 10% 27% fflN264 5) FAD25-3-5 22-27% 41-48% ÍBN264 6) FA025-3-9 6-35% 22-53% ÍBN264 7) FA029-2- 4 17-34% 32-50% pBN427 8) FA029-2-5 18-25% 35-42% pBN427 9) FA029-2-7 29% 46% pBN427 10) FA029-3-2 9-33% 25 -50% pBN427 5 25 - 20% 1 ») FA029-3-4 26-29% 40-43% pBN427 Typically, 20 grains per 4 ears of maize sisters from each event were analyzed based on the single grain. The range indicated the stearate content from the lowest to the highest from the single grain result of such an event. b. Total saturated fatty acids = 16 + 18: 0 + 20: 0. f = Purified fragment p = intact plasmid DNA. d. Co-suppression frequency = total number of events showing the total number / positive phenotype of the coarse-resistance clones generated from the respective transformation experiment.

The tansgenic phenotypes in the new events were determined by the composition of the lipid in single grains harvested from the fully mature maize ears using the same GC method described above. The sampling was not destructive, because only very small pieces of embryos were cut from individual grains and used for the fatty acid composition tests. The grains remain viable and can be planted either in the greenhouse or in the field for propagation of the next generation. Table 4 shows transgenic events identified with the high stearate phenotypes (and high total saturated fatty acids) phenotypes to the RO generation: 1. Typically, the lipid assay was performed on the 5-20 grains of each ear of corn, taken from corn ears 4-6, from sister plants for each transgenic event. The contents of total saturated fatty acid and stearate are shown as a percentage in oil, and the ranges presented indicate the lowest and highest percentages among all the single grains analyzed in the event. The results indicate that a consistently high frequency (10-20% of co-suppression events can be obtained in maize (Table 4 and 6), if an intact plasmid DNA or a purified fragment is used. Small portion of the DNA contamination vector may still be present in the preparations of the purified fragment, and Souther stain analysis can be performed to verify the completely free events of a bacterial selectable marker. of a DNA fragment tends to generate events with simpler insertion patterns (one or more copies of the transgene insertion), which use the intact plasmid DNA, the latter can form complex concatemers and integrate together in the plant gene when used in the biolistic method, resulting in a locus or complex insertion site, which may cause some instability of the transgene. c. Transgenic corn with a content of High Oleic Acid in Grains The corn calli were co-bombarded with pBN257 DNA (SEQ ID NO: 8) and a bar gene fragment, the transgenic maize plants were produced, and the RO: 1 grains were harvested, and the lipid composition was analyzed. as described above. Another transgenic event, FA014-5-1, was identified with a high oleate phenotype. Figure 6 shows a typical example of the segregation of R0: 1 seeds harvested from a single ear, and their corresponding phenotypes. The ear was harvested from a female plant of native type (LH132), pollinated with pollen from a transgenic plant of line FA014-5-1. The proportion of the positive phenotype: wild type = 1: 1, indicated that line FA014-5-1 contains a locus or single site insertion, and the high oleate transgene trait may be dominant. The lipid profile of the phenotype is, on average, 12% 16: 0, 1.3% 18: 0, 70% 18: 1, 15% 18: 2, and 1.4% 18: 3. The highest content of oleic acid found in the samples taken from this ear was 81 and in one of the ears the oleic acid content in some of the grains was 83%. The accumulation of high levels of oleic acid is at the expense of linoleate as shown in Figure 6. There is approximately 2-4% increase in palmitic acid, without any major change in the contents 18: 0, 18: 3 , 20: 0, or 20: 1. The R3: 4 grains of homozygous plants were harvested, with the composition of lipids as 10% 16: 0, 1.5% 18: 0, 68% 18: 1, 19% 18: 2, and 0.8% 18: 3. The result of the composition is similar to that of the heterozygote R0: 1 with a content of 2% oleate, indicating that the genotypic background can influence the transgenic phenotype. When transgenic R3 homozygous plants were used as the source of pollen, and crossed in the high oil inbred lines QX47 (which have a total oil content of 14%), QH102 (which have a total oil content of 9%), or a hybrid line 34K77 in the TopCross® grain production method (U.S. Patent Nos. 5,704,160 and 5,706,603), the respective lipid composition of the grains in each cross is shown in Table 5. The oleate content in the grains from the pure QX47 line is ~ 43%, and the crosses from FA014-5-1 with this line also resulted in a high oleate content in the grains (79% vs. 68% of the grains of homozygous FA014 plants). -5-1). The total oil content of the grains from the crosses of FA014-5-1 to QX47 is 8% -10%, and is 6% -7% of the crosses from FA014-5-1 to QH102.

Table 5. Grain lipid composition in RO: 1, R3: 4 momocigoto, and several crosses of FA01 -5-1 Genotype Phenotype 16: 0 18: 0 18: 1 18: 2 18: 3 R0: l xLH132 12 1J 70 15 1.7 R3: 4 the same "10 1.5 68 19 0.8 QX47 (HO) x R3 9 2 79 10 0.4 QH102 (HO) x R3 10 2 71 16 0.5 34K77 (TC) x R3 10 1 71 16 0.7 W7 15 18 65 0.7 a The grains were from the same homozygous R3 plants. The same homozygous plants were used as the source of pollen for the cross with the female plants listed below. b A few hybrid 34K77 plants were the same to obtain the native type grains as the control.

Using similar processes, new transgenic events with high oleate phenotypes were generated (Table 6). The trait gene constructs, which were used in these experiments are either pBN257 or pBN428 (Figure 7B). The transgenic sequence in the Sal I fragment (position 44-3468) of pBN428 is identical to the Sal I fragment of pBN257 (position 2-3426), except that pBN428 is using a vector structure with a selectable marker gene of resistance to hygromycin in pBN257. The trasngen prepared by the bombardment was either the digested restriction enzyme and the DNA fragment purified on agarose gel, or the intact plasmid DNA as indicated in Table 6. The restriction enzyme that was used to cut the transgene can be Sal I or Xba I, which releases a fragment of the transcriptionally functional transgene of 3.4 kb, and can be purified by agarose gel electrophoresis.

Table 6. Transgónicoß events with the high oleate phenotype Transgenic event Oleate "Conctructo b Co-suppression frec.c Native type -22% i) FA014-5-I -70% pBN257 1/10 = 10% 2) FA027-1 -9 60-69% ÍBN257 3) FA027-4-1 79-87% ffiN257 3 20 * 15% 4) FA027-4-5 81-87% ÍBN257 5) FA028-1-8 39-63% pBN428 6) FA028-MO 50-55% pBN428 7) FA028-3-1 64-78% pBN428 4 32 = 13% 8) FA030-3-3 30-83% pBN428 9) FA030-2-1 78-82% ÍBN428 10) FA030-2-9 82-83% fBN428 6/61 - 10% 11) FA030-3-1 80 -84% ÍBN428 12) FA030-3-3 40-68% ÍBN428 13) FA030-4-25 42-77% ÍBN428 14) FA030-5-I7 71-86% ÍBN428 15) F? 031-5-8 58 -76% ÍBN428 1/6 * 17% to. Typically, 20 grains per 4 ears of maize sisters from each event were analyzed based on the single grain. The range indicated the stearate content from the lowest to the highest from the single grain result of such an event. b.f = Purified fragment p = intact plasmid DNA, c. Co-suppression frequency = total number of events showing the total number / positive phenotype of the coarse resistance clones generated from the respective transformation experiment.

Two of the high oleate events, FA027-4-1, and FA027-4-5, were carried out towards the generation of Rl: 2. The oleate content of the grains of these progenies indicated a consistent high oleate phenotype. (81-87% oleate by single grain analysis). d.Transgenic Maize with High Levels of Oleic and Saturated Acids in Grains Corn with a high level of saturated fatty acid and a high level of oleic acid in the grains, can be produced by crossing a high saturated transgenic line (FA013-2-4 or FA013-3-2) and the transgenic line of high oleate (FA = 14-5-1), or by crossing the high saturated transgenic line with a high oleic acid mutant such as lines B730L or AEC2720L (W095 / 22598). An alternative approach for obtaining a corn plant high in both saturated fatty acids and high oleic acid is to create a transgenic line with a transgene construct containing the fusioned fad2 and delta-9 desaturase genes, such as in pBN412 or pBN431 (Figure 7C), or the transformation can be done by co-bombardment with both pBN257 (or pBN428) and pBN264 (or pBN427 or pBN262). The transgenic events comprise the chimeric gene of pBN431, possess a phenotype in which the total saturated level is not less than about 30% of the total seed content, the level of stearic acid is in the range of about 11% to 31% of the Total seed oil content and oleic acid level is in the range of about 27% to about 37% of the total seed oil content. It is believed that the oils can be obtained which have an oleic acid level in the range of about 35% to about 45% of the total seed oil content by crossing these transgenic events with a line having a phenotype of high oleic acid, for example, any of the transgenic events set forth in Table 6 above, or B730L or AEC2720L, caules are referred to above. The high oleic acid corn oil and high stearic acid resulting from such a transgenic fact, can be used in a combined or non-combined form as a mergarine or kneading butter, and can be combined with a palmitic acid fat to form a substituted of cocoa butter. clx? > Shep, Jennie B. 3. Z.? U Pont? E Nemours and Corapajiy < 12 D > GENES FOR DESATDRASAS. TO ALTER THE LIPID PROFILES IN THE MAIZE GRAIN < 130 > EB-1137-A < 140 > < 141 > < 150 > 60 / 0.18,987 < 151 > UHE 11, 1298 < 160 59 < 17D > Microsoft Office 97 < 210 > 1 < 211 > 1790 < 212 > DNA < 2l3 > Zea raavs < 40C > I cggcttctcc cctccctcct ccctgcaaat cctgcagaca ccaccgctcg tttttctctc € 0 c gg cagga gaaaagggga gagagaggtg aggcgcggtg tccgcccgat Ctgctctgcc 120 ccsacgcagr tgttacgacc tcctcagtct cagtcaggag caagatg gt gccggcggca 180 gsatgaccga gaasgagcgg gagaagcagg agcagctcgc ccgagctacc ggtggcgccg 240 cgatgcagcg gtegecggtg gagaagcctc cgttcacect gggtcagatc aagaaggcca 300 tcccgccaca ctgcttcgag cgctcggtgc tcaagtcctt ctcgtacgtg gtccacgacc 360 tggtga gc cgcggcgctc ctctacttcg cgctggccat cataccggcg ctcccaagcc 420 cgctccgcta cgccgcccgg ccgctgeact ggatcgcgc sgsgtgcgtg tgcaccggcg 4B0 tgtgggtcat cgcgcacgas tgcggccacc acgccttctc ggactact g ctcctggacg 540 gg acgtggt cetggtgctg cacteg cgc tc ggtgcc ctacttctcg tggaagtaca 600 gccaceg'gcg ccaccactcc aacacggggt ccctggagcg cgacgaggtg ttcgtgccca 660 agaagaagga sgcgctgccg tgg acacee cgtacgtgta caacaacccg gtcggccggg 720 tggtgcacat cgtggtgcag cccaccctcg ggtggccgct gta ctggcg accaacgcgt 780 cggggcggcc gtacccgc ct c cgecrgcc acttcgaccc cggcccc atetacaaeg 840 acrcgggagcg cgcccagatc ttcgtctcgg acgcsggcgt cgtggccgtg gcgttcgggc 900 tgtacaagct ggcggcsgeg ttcggggtct ggtgggtggt gcg gtgtac gccgtgccgc 960 tgctgatcgt gaacgcgtgg ctggtgctca tcacctacct g cagcacacc cacccgtcgc 1020 tcccccacia cg &ampC cgagc gagtgggací ggctgcg gg cgcgctggcc accatggacc 1080 gcgact cgg catcctcaac cgcgtgttcc acaacatcac ggacacgcac gtcgc cace 1140 acctettctc caccatgccg cactaccacg ccatggaggc caccaaggcg atcaggccca 1200 tcctcggcga ctactaccac ttegaecega cccctgtcgc caaggcgacc tggcgcgagg 1260 ccggggaatg catcCacgtc gagec gagg accgcaaggg cgtcttctgg t caa AAGA 1320 agttctagcc gccgccgctc g agagctga ggacgctacc gtaggaatgg gageagaaac 1380 ca39a-? SAS9 agacggtact cgccccaaag tctccgtcaa ectatetaat cgttagtcgt 1440 gacgggaaga cagtctttta gagatcattt gg cacagag acgaaggctt actgcagtgc 1500 gctgccatca catcgctaga agtacaagta ggcaaattcg tc ttagt gtgtcccatg 1560 ttgtttttct tagccgtceg ctgctgtagg ctttccggcg gcggtcgttt gtgtggttgg 1620 cafcccgtggc catgcctgtg cgtgcgtggc ^ cgcgc TGTC gtgtgcgtct gtcgtcgcgt tgg 1680 g CGTC tcttcgtgct ccccgtgtgt tgttgtaaaa caagaagatg ttttctggtg 1740 tctttggcgg aataacagat cg cgaacg aaaaaaaaaa aaaaaaaaaa 1790 • 21C > 2 < 211 > 1733 -c2I2 > DNA c21 - > Zea mays < 220- > «221 > CDS c222 > C17S3 (1351) c40D? 2 fccctccctcc tectccctgc aaatcgccaa atcctcaggc accaccgctc gttttcctgft 60 gcsgsgaaca ggagagaagg ggagagaccg agagaggggg aggcgcsgcg tccgccgsat 120 ccecgacgca ctgcteegac gcctgtcacg ccgtcctcac tctcagccag cgaaa atg 17.beta. Met ggt gcc ggc ggc agg atg acc gag aag gag cgg gag gag cag gag cag 226 Gly Ala Gly Gly Arg Met Thr Glu Lye Glu Arg Glu Glu Gln Glu Gln 5 10 15 gag cag gtc gcc ggc gct ggc ggc ggc gcg cea gtg cag cgg teg 27-í Glu Gln Val Ala Arg Ala Thr Gly Gly Gly Ala Ala Gl Gln Arg Ser 20, 25 30 ceg gtg gag aag ceg ceg ttc acg ttg ggg cag ate aag aag gcg atc 322 Pro Val Glu Lys Pro Pro Phe Thr Leu Gly Gln He Lys Lys Wing He 35 40 45 ceg caeg falls tgc ttc gag cgc tcc gtg ctg agg tcc ttc teg tac gtg 370 Pro Pro Sis Cys Phe Giu Arg Ser Val Leu Arg Ser Phe Ser Tyr Val SD 55 60 65 gcc drops sac ctg gcg etc gcc gcg gcg etc etc tac etc gcg gtg gcc 418 Jila HÍS Asp Leu Ala Leu Ala Ala Ala Leu Leu Tyr Leu Ala Val Ala 70 75 80 gt ata ceg gcg cta ccc tgc ceg etc cgc tac gcg gec tgg ceg ctg 466 Val He Pro Wing Leu Pro Cys Pro Leu Arg Tyr Wing Wing Trp Pro Leu B5 90 95 tac tgg gtg gcc cag ggg tgc gtg tgc acg ggc gtg tgg gtg atc gcg 514 Tyr Trp to Wing Gln Gly Cye Val Cys Thr Gly Val Trp Val lie Wing 100 105 110 falls gag tgc ggc falls falls gcc ttc tcc gac falls gcg etc ctg gac gac 562 His Glu Cys Gly His His Wing Phe Ser Asp His Wing Leu Leu Asp Asp 115 120 125 gec gtc ggc ctg gcg ctg drops teg gcg ctg ctg gtg ccc- tac tcc teg 610 Wing Val Gly Leu Wing Leu His Ser Wing Leu Leu Val Pro Tyr Phe Ser 130 135 140 145 tgg aag tac age falls cgg cgc falls falls tce aac acg ggg tcc ctg gag 656 Trp Lys Tyr Ser His Arg Arg His His As As Thr Gly Ser Leu Glu 150 155 160 cgc gae gag gtg ttc gtg ceg agg acc aag gag gcg ctg ceg tgg tac 706 Arg Asp Glu Val Phe Val Pro Arg Thr Lys Glu Ala Leu Pro Trp Tyr 165 170 175 gcc ceg tac gtg falls ggc age ccc gcg ggc cgg ctg gcg falls gtc gcc 754 .Ala Pro Tyr Val His Gly Ser Pro Wing Gly Arg Leu Wing His Val Wing 180 185 190 gtg cag etc acc ctg gsc tgg ceg ctg tac ctg gcc acc aa c gcg teg 802 Val Gln Leu Thr Leu Gly Trp Pro Leu Tyr Leu Wing Thr Asn Wing Ser 195 2D0 205 ggg cgc ceg tac eco cgc ttc gcc tgc drops ttc gac ccc tac ggc ceg 850 Gly Ar Pro Tyr Pro Arg Phe Ala Cys His Phe Asp Pro Tyr Gly Pro 210 ~ '215 220 ^ 25 ate tac ggc gac csg sag cgc gcc cag atc ttc gtc teg gac gcc ggc B98 He Tyr Gly Asp Arg Glu Arg Ala Gln He Phe Val Ser Asp Ala Gly 230 235 240 gtc gcg gcc gtg gcg ttc ggg ctg tac aag ctg gcg gcg gcg tcc ggg 946 Val Ala Ala Val Ala Phe Giy Leu Tyr Lye Leu Ala Ala Ala Phe Gly 245 250 255 etc tgg tgg gtg gtg cgc gtg tac gcc gtg ceg ctg ctg atc gtc aac 994 Leu Trp Trp Val Val Arg Val Tyr Ala Val Pro Leu Leu He Val Asn 260 265 270 gcg tgg ctg gtg etc atc acg tac ctg cag falls acc cae cec gcg ctg 1042 Wing Trp Leu Val Leu lie Thr Tyr Leu Gln Hie Thr His Pro Ala Leu 275 280 285 ccc falls tac gac tsg ggc gag tgg gac tgg cgc ggc gcg etc gcc 1090 Pro K ± s Tyr Asp Ser Gly Glu Trp Asp Trp Leu Arg Gly Ala Leu Wing 290 2S5 300 305 acc gtc gac cgc gac tac ggc gtc etc aac cgc gtg ttc falls atc 113B Thr Val Asp Arg Asp Tyr Gly Val Leu Asn Arg Val Phe His His He 310 315 320 acg gac acg falls gcc gcg drops falls etc tcc tcc acc atg cec falls tac 1186 Thx Asp Thr His Val Ala His His Leu Phe Ser Thr Met Pro His Tyr 325 330 335 falls gcc gtg gag gcc acc agg gcg atc agg ccc gtc etc ggc gag tac 1234 His Ala Val Glu Ala Thr Arg Ala He Arg Pro Val Leu Gly Glu Tyr 340 345 350 tac cag tcc gac cec acc cct gtc gcc aag gcc acc tgg cgc gac gcc 12B2 Tyr Gln Phe Asp Pro Thr Pro Val Wing Lys Wing Thr Trp Arg Glu Wing 355 360 365 agg gag tgc atc tac gte gag cct gag aac cgc aac egc aag ggc gtc 1330 Arg Glu Cye He Tyr Val Glu Pro Glu Asn Arg Aen Arg Lys Gly Val 37D 375 380 385 ttc tgg tac aac age aag ttc tagccgccgc ttgctttttc cctaggaatg 1361 Trp Phe Asn Ser Tyr Lys Phe 390 gagaaa gga tcaggatgag aagatggtcc tgtctccatc tacctgtcta atggttagtc 1441 tgcágtgcca tcsctagatc ctaggcaaat tcagtgtgct cctgtgcccc atggctgtga 1561 ctctcaagta gccttgggta gtcaagttet cttgtttttg ttrttagtcg cccgctgttg 1621 taggettgec ggcggcggtc ttcgegtgg ccgcgccttg tcgtgtgcgt ctctcgccac aaaaaaaaaa aaaaaaaaaa 1681 aaaaaaaaaa ctccccaaaa tctcttcgtg aa c210 >1733; 3 211 > 352 - «212 > PRT < 213 > Zea mays < 400 > 3 «et Gly Ala Gly Gly Arg Met Thr Glu Lys Glu Arg Glu Glu Glr. Glu 1 5 10 15 Gln Glu Gln Val Wing Arg Wing Thr Gly Gly Gly Wing Wing Val Gln Arg 20 25 30 Ser Pro Val Glu Lys Pro Pro Phe Thr Leu Gly Gln He Lys Lys Wing 35 40 45 lie Pro Pro Kie Cys' Phe Glu Arg Ser Val Leu Arg Ser Phe Ser Tyr 5C 55 60 Val Wing Hie Aep Leu Wing Leu Wing Wing Wing Leu Leu Tyr Leu Wing Val 65 70 75 80 Wing Val He Pro Wing Leu Pro Cys Pro Leu Arg Tyr Wing Wing Trp Pro 85 90 95 Leu Tyr Trp Val Wing Gln Gly Cys Val Cys Thr Gly Val Trp Val He 100 105 110 Wing His Glu Cys Gly His His Wing Phe Ser Asp Hie Wing Leu Leu Asp 115 120 125 Asp Wing Val Gly Leu Wing Leu His Ser Wing Leu Leu Val Pro Tyr Phe 130 135 140 Ser Trp Lys Tyr Ser His Arg Arg His His Ser Asn Thr Gly Ser Leu 145 15D 155 160 Glu Arg Asp Glu Val Phe Val Pro Arg Thr Lys Glu Ala Leu Pro Trp 165 170 175 Tyr Ala Pro Tyr Val His Gly Pro Pro Wing Gly Arg Leu Wing His Val 180 185 190 Wing Val Gln Leu Thr Leu Gly Trp Pro Leu Tyr Leu Wing Thr Asn Wing 195 200 205 Ser Gly Arg Pro Tyr Pro Arg Phe Wing Cys Hie Phe Asp Pro Tyr Gly 210 2 15 220 Pro He Tyr Gly Asp Arg Glu Arg Ala Gln He Phe Val Ser Asp Ala 225 230 235 240 e 245 250 255 Gíy Leu Trp Trp Val Val Arg Val Tyr Ala Val Pro Leu Leu lie Vai 260 265 270 Asp Ala Trp Leu Val Leu lie Thr Tyr Leu Gln His Thr His Pro Wing 275 '28C 2S? Leu Pro His Tyr Asp Ser Gly Glu Trp Asp Trp Leu Arg Gly Wing Leu 29D 295 300 Wing Thr Val Asp Arg Asp jr Gly Val Leu Asn Arg Val Phe Hie His 3D5 310 315 320 lie Thr Asp Thr His Val Wing His Kis Leu Phe Be Thr Met Pro Hie 325 330 335 Tyr Hie Wing Val Glu Wing Thr Arg Wing He Arg Pro Val Leu Gly Glu 340 345 350 Tyr Tyr Gin Phe Asp Pro Thr Pro Val Wing Lys Wing Thr Trp Arg Glu 355 3ED 355 Wing Arg Glu Cys He Tyr Val Glu Pro Glu Asn Arg Asn Arg Lys Gly 370 375 3BD Val Phe Trp Tyr Asn Ser Lys Phe 385 390 < 21D > 4 < 211 > 12313 < -212 > DNA < 2l3 > Zea maye < 400 > 4 ttgtgatgtt gtcagggggg cggagctatg gaaaaacgcc agcaacgcgg ctttttacgg 60 ttcctggctt ttgctggctt ttgctcacat gttctttcct gcgttatccc ctgattctgt 120 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga 180 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc 240 cgcgcgttgg ccgattcatt aatgcagct gcacgacagg tttcccgact ggaaagcggg 300 cagtgagcgc aacgeaatta atgtgagtta gctcactcat taggcacccc aggctttaca 360 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg 420 atttaggtga cactatagaa aaacagctat gaccatgatt acgccaagct tactcaagct 480 atgcatcaag cttggtaccg agctcggatc cccttgcagc agagagcaag ttccaacaat 540 a.ccccCaaec acccaccatt cattgcatcc acttcceaca aagttttcta acttacaaga 600 gctatagcat tcaatacaag acacaccaaa gagatcaaat cctctcccaa gtccatagat ca -660 ttccaat caaataatga ctagtgagag ggtgacttgt gttcatttga gstcttgcgc 720 ttggattgct tctttttctc attctttctt gtgatcaact caattgtaac cgagacaaga 780 gacaceaatt gtgtggtggt cettgcgggg actttgt tc tcgtttgatt gagaagagaa 840 gcteactcgg tctaagt gat cg ttgagag agggaaaggg ttgaaagaga cccggtcttt 900 gtgaccacct caacggggga gtaggtttgc aagaaccgaa cctcggt aa acaaatattt 960 tgcttacaat ttgtttttcg ccctctctct cggactcgtt aatatttcta acgctaaccc 1020 ggcttgtagt tgtgcttaag tttataaatt tcagattcgc cctattcacc cccctctagg 10B0 cgactttcag taccgttata tatgctttcg atttatcctg cccctaagtc agttactaga 1140 sagattgata ttcttaggag gcgtcttctt tggcaagggg gtcgtcagtc caaaaaaatt 1200 catttágttg attggttgtc ggtgtgctct cccaaaaagt cagggaggtc tgggtgttct 1260 gaatctegat tttatgaatg attecttaat gactaaa gg ctttgsaata ttsaaacttc 1320 tggcsa gaatggccca & aaa ctattaccag taaatatatt aagggaaaac cccttatttt 1380 caa gatcaa agacaaggrg attcac ctt ctggaaaaaa aattctgagt ctgcgtgata 144C atttttacaa attttgcaaa tctggggtgg gaaacggttt gaagactagc ttttggaaga 1500 gtatctggat tggaaatctg cccctgtcc tfccagttt.cc tgttctattt gacttgtctt 1560 gacaaaga cattacsgtt aatgatgtca tggcttctaa ttttgaggtt cttacattta 1620 gaagaaggat tgttggtaat ctgagggttc taatggatga gttggtgagt tgttgcaatc 1680 atgt ttctt gtctgacca g gaggacagaa ttgtgtggag tccggggaga aaaggctttt 1740 crtattaattt tatttaaaaa agaaaatggc agatcaagtt ttgatttcat ataagttctt 18D - gtggaaaatc aagattttca tg gttggt tgtgagaaat aaaattctta ctaaagacaa 1B6C gga tgt tatgaagaaa aggaactgga atggttcttt gt ttctgtggcg tggatgaatc 1920 cattgatcat ttattctttc attgtcccat tgcgagatat atgtggagag tgattcaagt 1980 ggccttgaat ctgaggatga ttccaagtag tattagcaac ctttatgaca accggttatg 2040 tagaceaaaa gataatattg ctaatctggt tttgtttggc tgtgsageta 210C tgctctgggc aatttggcgc actagaaatg attggtgctt tgggaataaa actatgcttg atccctctaa 2160 catcattttt ctttgctgct tctggctgga ttcctaggct attcg gaaagaagga shits 2220 gcaaaaaata gtggtccaag gaagcaaget aatctgaaag acaacaagtg aagcattcag 22B0 cegagegttt gggtggtgcc cg agacag gcgtatttct ggttgatctt aagctggaac 2340 ttgaatgatg gtgctggtgc tatcctttct tttggtggtt gtcttggttc agtatctctt 2400 tctgtca gttgcaccag ga tatgattgta aataagaaag gcttatgttg ttaatcgtaa 2460 gtcaaacttt attcgctatc ataggtcctc cactgatcta gtttgatagt gttaggagtc 2520 tagatagaga tctgaccttg ttcgattttt tt gtttatt ggtcgca tga gtactgttgt 2580 ttcaaacttt eararttett aatgaáatag ggcttcgcc cctacaac c tgatcacttt 2640 cactrgsata cgggagacct c ccaattca tactgtgtgt tggggggggg ggtggggggg 2700 AATAA ACAA cgagagaaaa aaatctgagc tttaccatta cagaggtcag aggttacgaa 2760 caccgtcaaa cagctgcatc atgcgecagt gcacccacgt cctgttggat taatgtgggc 2B20 ttaatattca ttggcccaaa ataatagtca atgctaatgg cccaotttaa tgctatggtg 2B80 tactaattat ttagtaccat attggaagtt caaaggacaa atcaatcaac ttaaataggt 2940 ggaccattgg tgcatctatt gagaag tga gaaaagaatg aaagactgcc acacgcgcgc 3000 gcgcgccgce gccgccggcc gggcccgtgg ccgtggccgt ggccgtggct cgtggctcgt 3060 ggtagategg accttggtcc gaatattcct ttcaaacggt tgtgcatttt gcctggattg 3120 atgaccgtca taataaccgt ctgtttcctg tcttatggct agtaacggac gtcagttact 3180 g cg cagtt tccagttcta atgcgcgacc gtttctgtcc gttgctcttc tcccttcttc 3240 taagaatgga tgaccggcta gagggagagc tcttecagtc aggcgaattt atctcacgcg 3300 aattgcaaac aacacattcc ccgtcccatc ttctgcgagc acagagagag tgggagagca 3360 ggcctccgaa atcaccgacc gcagagatac acttgcacgg gtgtgcgggc ga t tcagat 3420 ttggggagcg tcttcgcgac tgctcgcgtg atcgtccaca gc gctgtt cgtcgcctac 3480 ccaagttgac gcgtgctgct gttcttcttc ccggcgaccg ttcgagggac tgcactgegt 3540 geacegactt acaccttcet catcgaacaa cgtacgacta acacacgaga tgtctcgtgt 3600 gaatggagcc actggtgcct tgagcatcgg tccctccgct gggtacaetc tgttcttcgt 3660 atttgtgcat gtttcattgc tgtttactgc ttatgcgagt -ag tatacac acatgcacat 3720 acatg catc acatatatca ctggattaaa cactgatttt ttaaaactaa aaatgcctaa 3780 ctttctaaca cgtccgagca tcaccgcttg cttgcgccct cggcggtctg gaatctgcat 3840 gtcgccgggc gcggggcgcc ggcgeaccgc ccecgccgtg gtctgctacc cgtcagtccg 3900 cgccacactt cttgaggaga acatcg cgc acgcgggcac geggcgtggc cggcggtgac 3960 aactgcagag catgg cgcc acttgtcagt tctgtcagca agggtgccgg tgccagtgcc 4020 ßgcaccgagc tcgctttgtt tgcctgctgc cagtgtggca gacatcggac gacggagctg 4080 taggcgccat gcgeatacta gatgggtatc tttcggtgct tggaacttgg ttcacaggtg 4140 gatgtct ca tgcacatcgt ctctacagtc tacactgaat caagcacacc attacaccaa 4200 tgeatttttc tgttgcct t atggagatag ctgattagtt caccgaa ga agcacac caa 4260 tttccaacca cgtgcgtact gttgcgcttg gagatagctg etggttagtt caccgaatcc 4320 gcggcctaac tccggacaea tttttttctt ctggtagatc gcatcacatg cttgctcccc 4380 atcacgggct gcaaggtgcc acccctcgct gsctgt cca ggccatcaac accgtgggtt 4440 tggcaaccgg tgttgcgcta cccaatgcct gagaaaaatc gtggtacggc ccaaccatgg 4500 aagatcagcc aaaatgagct cacatgaaac tgcccaaaac aggaagaggg tagttgaaat 4560 aaaatgggtt cagtgacggt acgaagtcag atttgaagaa gtgcccaacg ataatacata 4620 attcgtatta gttcaactac tttttggaca aatcttcagg tcceaaatta tttagttcac 4680 cgctgcaaac tactatatgg aaagatacga cgatcaatca aaaggcaatt ttctttggtg 4740 aaccaatcgt ttcacaaggg aaatcaacta cgcegatg c tgctgttttc cttagggcct 4800 gttcgcttct tcaggaatga acttggattc attcgagctc atcaaaattt atataaatta 4860 GAAAA at ccggctaaga actattecag ggctccaatc cgtgaaaacc gaacagagcc 4920 ttagagagcc cg ctg gg ataggagtat atagcttttt gtttaagctt ttttttcaat 49B0 ttcrgatcac cagaagatgt cgcaaaactg ttaaac ct aactttttaa cctgttccta S040 taagaatcat tttagtcaaa attatctaaa atcaatatga ggacagaatc aaccgagtcc 5100 t tatgaaaac cgtcattttc tatatcctaa atcacataaa etattttatc tttcttcaca 5160 C ttltCEÍC stgaaactgt attoeetaca aecatatttt tctggcagte agattctaaa 5220 aaaaatcctc acaaaaaagt tgaaceaaae tcgegageca cgggccegcg tccggcgctg 5280 cacga ctgt gtcacgcctc ccggcctcec ggggtccagc caaatagggc tctacatgtg 5340 ggcca caca gatttcacgt ccgccgacgt ggttacsgcg tcacatgatc acatctggct 5400 cctccgggcc caggcgccag tgacgccgtg cccgcctcta aatagcgcct ctctcccggg 5460 ctgceoscgg aaccgaggcg gtcasgctcc ctcctccttc ctectccctg caaatcctgc 5520 aggcaccacc gctcg TTTC ctgtccggg gacaggagag aaggggagag accgagagag 5580 ggtgagscgc ggcgtccgcc cgatctgctc cgccccccga agcagcctgt cacgtcgtcc 5640 tcaetctcag gtacccgcat ttagccttce tggattgtta tggatc cta gtgccecccc 5700 tgccactgtt ccatagattg ttcegaatgg attggtgagg aatcgaccgg cgttcggttc 5760 tgggttgctg agcccggcaa cgggctcgtg gecggccgtc gattcggcag cggcactcgc 5820 cg cgcgccg cccggtcggg tcgggtcggg tc ctgcaaa ctcgccgtag cgcctgccsg 58B0 tcgagctttg acaecgacct caccggcggg catccgcggc cctgccgatg tggatttcag 5340 gttttgeccc gatgaa cca cgcttgttcc tcaccagatc tgtaggtatg attcagcgag 6000 tggtgc catt cagatatttt gcccgtgcaa tgggaccgtg attgatctcc gcacctcctg 6060 ccgtgaccac tcgttttggg aacatggcat gccaccttta gecacgccea cgagctgacg 6120 agctcttcgc agctcccgta caaaaagctg caacctttgc aggtctttga ctccaaaggc 6180 ggcctctttg tttcggcgct cgcccccctc catgttgggc atgatgcgtt gcacttggtg 6240 cccgactcct ctgttttcta sctcctaatt ttttttgctg atgetactat agtactatta 6300 gctaagcgcg gagttggcga tgactgcgct caagaatcga gggcaatcga cctttggctc 6360 tgeatggacg aagccactrg tttttttttt ctttggtcat gtttttgaca tgcgaaactg 6420 cgaaggtggc agagtaggtg gatctttctg tctatgtttt ggccctactt gagaggaaga 6480 gacagtcgcs accgtgcaaa gtcccaaagg catccgacgg tggcgcgtcg atcgttacga 6540 ACAA tgcca gcaccacgga acgagccacc teccccgcgg ccagcccgcc atcgagcagg 6600 gattgaaaga tcacgactga aacgcgatct aatttttgtc ttttettttc stgtcccaag 6660 ttccttaata cttgatacgt gagctctata acactagagg ttttccattc ggaaaaatat € 720 gttcgctaaa gtcggtctct gattaaagtc ggctgcttga cggctgcaac tgtaatttat 6780 aaaacaaaaa gtaatttatt cactgtgttg acaecttttt atcacttcaa ggaggcgcca 6840 aagaaccggc ggcggtgctc tctggtcagg egcggacggt ccgcggcaca gggccggacg 6900 gtcegcgacc tggcgtgagg cggcggtgct c ct g cag gcgcggacgg tccgcggcac 6960 agggccggac ggtccgcgac ctggagcagg agctcgggtt ccctgcctga cggtcggacg 7020 gtccgcgcgt ge caggggc ggsggaagat cgccggcggc gcctggatct cgctcccggg 7080 agggaccccg tcggggagga gagatcctag gagttgtcta ggctcgggcc ggccgaccta 7140 gactectcta atcgacgtag agtcgaggag aggcggagaa tttggggatt ggaataetaa 7200 actagggcta aactagaact agactagaac tactcctaat tgtgctgaaa ataaatgcga 7260 gatagaagtg gtattggttc gat gttggg ggttcaatcg gccgtatccc tteatctata 7320 taaaggggga ggtccggatc cgcttccaac tgatttccga gttaatcccg cggttttagg 7380 t acaaatcc cgegagaaac taggaaccct aactgactct gcgcacgcgc ggaccgtecg 7440 cgccaccacc gcggacggtc cggaccgcgg accgtccggc ctccgggccg gaccgtccgc 7500 acggtcattt tgggttccaa catatgcccc ctgccttttg gtgaaggtcg acaaaccaaa 7560 agcattgaac taaacctgat GTAAG caec ggcttttcga tatggagatt attcaataaa 7620 gcaccaatat aaaggccgtt tcggattgta tctttctcgg ccatgaccat ttgatcaatg 7680 gatcaaaagg aatag aatgg aggtgccccc cagtctggat agacgaaggg actatacatg 7740 taccatggat tcatcatcgt gccattccat gtttgaacag gataatatae cgacgatgag 7800 taaataggtg gaaagtaccc tggtctcata gaatgaatag gcgatgcttg tgtgtcgcc 7860 tttcgggccg tctttgttta accgttttgt tttagcaggt ggctggggtt tctttgttga 7920 ccgatcacgt ggaacagtct ttttgctagc atttttggag to ca ctgat caaaagtagg 7980 atcggc tg atcagccgat tatatgtgct ttgaccttge gcctttttcc ttgctttgtg 8040 tagaggttga cgcctttggt cataggggga ctgtccggct gagttagccg gaccgtttgt 8100 ctgagcaccg gatcgtccgc ACGTA gtgc cggaccatct acgatgttcg ggctaggctg 816D atgtgt tgg ttcattaacr gtgcctgece cccagtgtct tcggactttt tagccttctc 8220 gtccsaaßcc tttcgagcaa tctctttttg tgatatatct gacatgcggg satcgccaar 82B0 gacgataccc ttgcctttgc ctttatcggc catttcgggc cctttttgca cgaaccaaga 8340 tgtgagttct attgtattaa caggaaaggg ttgtgtgtct aettgeatct cctgaaaagc 8400 caatcgacac tcatttatgg cegattgtat ctgtcgacga aaaacattac aatcattggt 8460 ggcatgagaa aaggaattat gec cttacá ataagcacgc ctctttaatt catcaggagg B520 ßggaatagta tgagttaatt taatgttgcc gtttttcagt aactcg caa 8580 atattttatc gcatttggca acactaaacg taaacttaac ttcttcttgt cgattctttc gaatcgactg 8640 taaagcagaa cacggtgaag gt tagcetg ccctggccaa accagttcag cggtgtatac 8700 acecgtggat tcatcgtccg agttatcata teccactaga tgeatettat ggctagccga 8760 ttttgatgtt tccttacttc ggctttcaca tgtcatagcc cgctggtgca agtgcactag 8820 cgaaaagaát tgggtaccat ctaatttttc ttttaagtag gg cgcaacc cattgaaagc 8880 tagccctgtt AGTT ttttt ccgegacatg aatctgaaag catcggtttc tagtgtcccg 8940 gaatctccgg atatagtcat taacegat and ttcaggcccc tgtcggactg aggctaagtc 9000 agccaattct aattcatgtt ctcctgagaa gaagtgttca tgaaatttct getetaatte 9060 ttcccaagag ttaatagagt ttggtggcaa agttgcgtac catgcaaatg cagtatcagt 9120 gaaaataaac aagggacaac gaacgcggta ggettcccca teagecaatt cccctaagtg 9180 tgctatgaat ggctaat & T gttcgtgtgt gct ttccca ccttcaccag aaaacttaga 9240 gaagtctggt attctagttc cctgtggata tggcacggtg tcgaatcggt ggctataagg 9300 cttccgatac gattgccctg tacctgacaa actaacaccg agtttgtccc tgaacatccc 9360 ggctacctcg tctctgatcc tctccgccat atctggcgac catctattga gtttgtgggt 9420 ggaaacctca ggttgcctga cateaetetg tcggctttcc tcccagggat gtttgaggtg 94B0 tgcgctaggt gtcctattaa tgtcaccaac tcctgcccta tacctctcag gctctet gt 9540 ggccgaacag tattcaaccc tatgtccatg aggtggtatg gcatgattat aatgtg ac 9600 TGGT gggca ccataatgtt gctgcgatga atgcggaaac tgtgcgtatt gcgcagctct 9660 cggctctgcg tatgcatatc cggacggtcc ggcataagag gccggatggt ccgcgacctg 9720 gccaaatggt tcgaaggtat atccggattg tccagccgta tatggtgcta catgggtagt 9780 ctcgtaecca gaccgtctgt cgtagataac tggacggtcc gcgatcgggc cgaatggtcc 9840 agggctgtac ccggacggat cggtcatata tggcgcgact tgggtagtct gcgcgtggac 9900 tcgtgtagag tcggatggtc cagcgtaata cgteggccgg tteatgecat atccggacgg 9960 cccggcgtaa gatggcgaac ggttcgcaac atatccg ac ggtccggcgt gatgeacegg 10020 acggtccgtg a ggggcega agt gttcagg gttgtaccct gatggtccgg cca-tgtacgg 10080 tgcgacct to gtgttttgcg tgcgggcctg catctggtcg gacggcccgg cgaaataege 10140 cggacggtcc gcggtataac cggactgtct gagatgatgc tcggacggtc cggtcgcgtc 10200 egtgtgccta cagtacctgc gtggttgcgg ctgtgatggt gacacgaaag cgtgcatcgg 10260 cataccatat gatggctggg tcaagggtga cccgtttata gccgatgtgt ttg cgt gg 10320 ttagactctc tggcactgta gttaggagca gtgatggaaa gctgatttct cgtatgcacg 1O3B0 taaatgcatt ttaatagatt catetacata ttgctttaat tgatctcctc gttgatccat 10440 gaaagtegta agagataggt ccggagtact cacagcggga ccctgaagtg gaggtaggag 10500 agattccata tcgatc ccc cttgacggac gatcttctgg tggcgatcta ccgtgaagtg 10560 tgacaagtac ttgtctgccg cctccttgcg cctttcggag agtttatgca gtagttccgc 10620 ctcttccttg tcgtgttgtt cgttccattg ccgcatcacc tccttctcat cgcgcattac 10680 gaggtettca aagggccttt ggtcatcagc egggagcgcc tccacggeeg gcttgatgat 10740 gttggtagtg gagatcttgg tgtgatccct agaaccggcc atttatgggc cgatttttgc ÍOBOO agattagaca cetagtcccc agcggagtcg ccaaaaagta cgttgacacc tttttggagg 10860 tgcaatcact tcaa aagaac cggcggcggt gctctctggt caggcgcgga cggtccgcgg 10920 cacagggccg gacggtccgc gacctggcgt gaggtggcgg tgctctctgg tcaggcgcgg 10980 acggtccgcg gcacagggcc ggacggtccg cgacctggcg tgasgtggcg gtgctctctg 11040 gtcaggcgcg gacggtccgc ggcacagggc cggacggtcc gcgacctgga gcaggagctc 11100 gggttccctg cctgacggtc ggacggtccg cgcgtgegca ggggeggcgg aagategeca 11160 gcggcgcctg ga ctcgctc ccgggaggga ccccgtcggg gaggagagat cctaggagtt 11220 gtttaggctc gggccggccg acctagactc ctttaatcga cg agagtcg aggagaggcg 11280 gagaatttgg ggattggaat actaaactag ggctaaacta gaactagact agaactaetc 11340 ctaattgtgc tgaaaataaa tgcgagatag aagtggt tt gg TCGA tg ttgggggttc 11400 aatcggccgt atcccttcat cta ataaag gggagg ctg gatetgette caactgattt 11460 ccgagttaat ccagcggttt taggtaacaa atcccgcgag aaactaggaa ccctaactga 11520 ctc gcg gcacggtcat tttgggttcc aacacacggt ataaacatta gaaattggta cggattaa g 1164C ctaascgaac ageCta aga ctgtgagcgc tcgaatccca ccttgtggga gcaccggagc 11700 acatgtgcag cttcgagcca tactggacgc tgcactgaaa gttttggcat tcatatagta 11760 aacgtccgtg gtcgacaggc acccácggcc t gaacatag cgatggaagt catggatcga 11820 tgagtca cgaagctga tt acaccgaagt cgattgacaa aggctatcta ccacgacatc 11880 sgaagacgtg aaatcgcaca atgaatagca gagggtaaaa ggtagagaga gatgtagcag 11940 attggttttt tgatgattga aagagtcgac cgtgttcatc tgatat cgt agasgtggíg 1200C gtcttatctg agcttccaca tgctscgatc gatttgttgg tccccatctt gctctcccac 12060 acaggaatac tattaaccat gttcag caa gaaagtgatg cggtegtgca cggcacatgc 12120 ca ctttgtg ggagccgcce ctaaccctcg ctgaatcagt cagtagtgcc aacttgctag 12180 agttttttct cctettgttt tggttcactc gacagatttt tgtttggatg agatcgctgc 12240 aacattgt c ttgatccaca cttgcctgat c accgtct cgttcgtgtt cgtgccagca 12300 accagegaaa atg 12313 c210 > 5 < 211 > 2907 c212 > DNA < 213 > Zea mays < 40D > 5 caggtacccg cattagcctt cctctattct ggatgatccc ccctgccagt gtttcataga 60 ttg gaa c tggattgatg aggaatcgac cggagtttcg gttttgggtt gctgagctcg 120 gcagcagggt gacagttegc cgtagcgggc tcgtgaccgg ccgtcgccgg cggggcgggt 180 ccggccgagc tcgrgtcgtc gatccgtagc gttgggtctg ggagaaagta atgggatgcg 240 gccgaactcg ccgtaccccc cgccggtcga gcttg catc gatctcaccg gcgggcatcc 300 gcacaagcct tgcgctgccg atgtggattt gcccagatta atcctggcaa agcgcgcttg 360 tttcccatct catcagatct gtaggattca gcgtggggtg ccgatcagat attttgcccg 420 tgcaatggat ccatgatctc tgccccctcc tgcccactcg tttcgggaac atgacatgcc 4B0 acttttggcc acgaactt t egcagctccc gtcaatcttg tgggtaaaag ctgcaacctt 540 tacaggccta gcctctttct ttatgcg tc ggtccctcca tgacagccat cgctgcgcct 600 gcgccctccc catgatggcc aaetgctccg ttgttctatc ttctgatttt tttactggta 660 ctattagcta agcacggagt tg cg caat tgcacccaag aattgactga ccttttagct 720 ccagcaattg ctgtgtctag gaagcaactc gttctgcttt gg cac cat aaaaaatate 78D tacttgtcca gatgggaaac cgtatatgct tttctaggaa tttggataga aaaaaataga 840 ctcaatccca gcgcgctcct gtcatcacac gctcgaggts gagggcagga aaccgccggc 900 ggcggcggcg gcagcgggga tggggagctc gttccgtggg tcttgtctgc ttgacctaga '960 aaacggcatc gtgatgaasg acgcgctacc gtccgatgcc ttgggatttt ggacggtggc 1020 gactg c cc tcccasgtgg 'ccacgtacag tcaaaaaccg agacagaaaa agattteace 1080 t actccgcct caccttcggc atgggccggc ggcatgtcag ggctctgcag ctgtgtctgc 1140 gcaacggtac aagacgccgc 335gstcgca gcctgcaagg ccggcaccga attctaggcc 1200 gcatgcaaca ccacatgatg ccggtgcaca gacacgatta gatatttttc tccagccgta 1260 gaataactcg gacaagtgtc gagaggcgtg gactagcaga tctgggtsca gttggcccct 1320 ctggtgacca gagtgacccg tccttcacct tggcgtggtc ggctgcaact cgctgtccga 1380 tgcaaattgc tgctactgct atgtccatgg catggagtcg catgtgccat ttcttccctg 1440 tttgtttggc tctccccgcc gtccgatcag aaagttaggg agacaattta ggccctgttc 1500 ctatctcgcg ag taaactt tagcagcttt tttttagcta ct ttagcca tttgtaatct 1560 aaacaggaga gctaatggtg gaaattgaaa ctaaacttta gcacttcaat eata aget 1620 aagtttagc aggaagttaa agtttatccc gtgagattga aacgggcctt tagacgggcg 1680 gcccttgtct tgtcagaatt aatgcacagt atcggcacgg cggccaagca tega tête 1740 cggatctggt ttctgtctcc atctgtgggc gccatggttg gctggtcgac aggacgcgct 1800 tgtgtcattt gggccaagcc ccaagggaga cagataacat ccgattccac ctcgtgegag 1860 cacatgtgcg gcttcgagec ataccatacc atactgaatg ccgcacttcc aaagttttgg 1920 catcaetgat aa attttggtaa cgcccaa caaacagaca caagatgaag atgaaaaacc 1980 ggatctt c taagatttat actaatgcgc cgtgcatctt ttacgttgct atatggtgct 2040 tcactaggct ttatcgtaaa ccgaactgat ttaccaccac cttcaatgca caaggcagag 2100 cacctgccat cttacgctga tttttttttg aaatatggtg tgcctctagg ctctggactg 2160 gtaggtgggt ttgeatg ag aaaagatgac tgggagctc atgcttgcta gcttgtcaaa 2220 to gaccact tetacegatg acgcaa att gccttgctct gtatggctat tggatagctt 2280 agatttgacc atatatggta gtactaccat ttatttttcc ttccgctgaa tcacctcaac 2340 gcacgttctt ggcgctgccg cttgttagtc tctcctgcct gctgctttcc attggtccag 2400 aagtcccttt cacaaatcac cgtccaattg cátgcagtac ac tc catgtt caaggscr 2460 gtt ttggac cagttcgttc aatgtaacat cacaagcgac aggacct aa tctgttttct 2S20 cttatttaa tgtagatttg ccgtagggtt ttgtaccatc cttggtcttg ctgtaaagtc 2580 tgcattttat tagttctgtg tggtggtaat .cagaattget ggtttgggct cgcacatgc; 264C gtgatcccca 'acttgctgtg gcgtggtagt tggatcgtgt ttaggcaaga aagtaaat c 2700 gatcatgcac cgcatatttg ccaccttcct gggagacgcc ccstcgtgcc gtgatctgtt 2760 ttactttggt tgattggtgg ccttte cgt ggttcacgtg acagcttttc tgatgggatg 2820 agatcact cgc tga gc aatgttgttg cttgattcac tcttactgta gcgtacttcc 2860 2907 tcgtttgtgt cagtcaggag caagatg«.210: > 6 < 211 > 18 «212 > DNA < 213 > ARTIFICIAL SEQUENCE < 220 > c223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OUGONUCLEOTTDO < 400 > 6 gayatgatha engargar IB < 210 > 7 «311 > 17 < 212 > DNA 213 > ARTIFICIAL SEQUENCE < 220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OIIGONUCLEOTIDE < 400 > 7 ccrtcrtaca tnagatg 17 c21Q- B < 211 > 1714 < e212 > DNA < 213 > Zea maye < 220 > < 221 > CDS < 222 > (134) (1312) < 400 > 8 ggeaegaget cactgccatt tgtttggttg ttcctctcgc tcgccagtcg ccaccaggca 60 gcaggcatcc caatctcgcg agagecagta gcggcggcgg cgcttccggc ttcccttccc 120 attggcctcc ggg atg gcg etc cgc etc falls gac gte gcg etc tgc etc 169 Met Ala Leu Arg Leu His Asp Val Ala Leu Cys Leu 1 5 10 tcc ceg ceg etc gcc gcc cgc cgc cgc age gge ggc agt ttc gtc gcc gcc 217. Ser Pro Pro Leu Ala Ala Arg Arg Arg Ser Gly Gly Ser Phe Val Wing 15 20 25 gtc gcc tcc atg acg tcc scc gcc gtc tcc acc agg gt gag aac aag 2 £ 5 Val Rla Ser Met Thr Ser Ala Ala Val Ser Thr Arg Val C-lu Asn Lye 30 35 40 aas cea ttt gct cct ceg agg gag gta cat gtc cag gtt here cat tea 313 Lys -o Phe Wing Pro Pro Arg Glu Val His Val Gln Val Thr His Ser 45"50 55 6C atg cea tct falls aag att gaa att ttc aag tea ctt gat gat tgg gct 361 Met Pro Ser His Lys He Glu He Phe Lys Ser Leu Asp Asp Trp Wing 65 7th 75 aga gat aat atc ttg here cat etc aag cea gtc gag aag tgt tgg cag 409 Arg Asp Asn He Leu Thr His Leu Lys Pro Val Glu Lys Cys Trp Gln BD B5 90 cea cag gat ttc etc cct gac cea gca tct gaga gga ttt cat gat gaa 457 Pro Gln Asp Phe Leu Pro Asp Pro Wing Ser Glu Gly Phe His Aep Glu 95 100 105 gtt aag gag etc aga gaa cgt gcc aag gag atc cct gat gat tat ttt 505 Val Lys Glu Leu Arg * Glu Arg Wing Lys Glu He Pro Asp Asp Tyr Phe 110 115 120 gtt tgt ttg gtt gga gac atg att act gag gaa gct cta cea here tac 553 Val Cys Leu Val Gly Asp Met He Thr Glu Glu Ala Leu Pro Thr Tyr 125 130 135 140 cag act atg ctt aac acc gac ggt gtc gtc aga gat gag here ggt gca 601 Gln Thr Met Leu Asn Thr Leu Asp Gly Val Arg Asp Glu Thr Gly Wing 145 150 155 age ccc act gct tgg gct ggg tgg acg agg gca tgg act gct gag gag 649 Ser Prc Thr Wing Trp Wing Val Trp Thr Arg Wing Trp Thr Wing Glu Glu 160 165 170 aac agg cat ggt gat ctt etc aac aac tac atg tac etc act ggg agg 697 Asn Arg His Gly Asp Leu Leu Aen Lye Tyr Met Tyr Leu Thr Gly Arg 175 180 185 gta gat atc agg ca att gag aag here att cag tat ctt att ggc tct 745 Val Asp He Arg Gln He Glu Lys Thr He Gln Tyr Leu He Gly Ser 190 '195 200 gga atg gat cct agg act gag aat aat cct tat ett ggt ttc gtc tac 793 Gly Met Asp Pro Arg Thr Glu Asn Asn Pro Tyr Leu Gly Phe Val Tyr 205 210 215 220 acc tcc tcc ca ga ga gcg acc ttc atc teg cat ggg aac act gct 641 Thr Ser Phe Gln Glu Arg Wing Thr Phe He Ser His Gly Asn Thr Wing 225 230 235 cgt cat gcc aag gac ttt ggc gac tta aag etc gca caa atc tgt ggc 88 ° Arg Eis Wing Lys Asp Phe Gly Asp Leu Lys Leu Wing Gln He Cys Gly 240 245 250 ate atc gcc tea gat gag aag cga cat gaa aet gcg tac acc aag atc 937 He He Wing Being Asp Glu Lys Arg His Glu Thr Wing Tyr Thr Lys He 255 260 265 fft39a9 * »9 tfcSf t?; T 9ß9 * c gac ect gat ggt here gtg gtt gct ctg 985 Val Glu Lye Leu Phe Giu He Asp Pro Asp Gly Thr Val Val Ala Leu 270 275 2B0 gct gac atg atg aag aag aag atc tea atg cct gcc falls ctg atg ttt 1033 Wing Asp Me .Met Lys Lys Lys He Ser Met Met Wing His Leu Met Phe 2E5 290 295 300 gac ggt cag gac gac aag ctg ttt gag falls ttc tcc atg gtc gcg cag 108.

Asp Gly Gln Asp Asp Lys Leu Phe Glu His Phe Ser Met Val Wing Gln 305 310 315 «SB ct ggc gtt tac acc gcc agg gac tac gec gac att ctt gag ttc 1129 Arg Leu Gly Val Tyr Thr Wing Arg Asp Tyr Wing Asp He Leu Glu Phe 320 325 330 ctt gtt gac agg tgg aag gtg gcg gac ctg act ggt ctg teg ggt gag 1177 Leu Val Asp Arg Trp Lye Val Wing Asp Leu Thr Gly Leu Ser Gly Glu 335 340 S45 gg aac asc gcg cag gac tac etc tgc acc ctt gct tea agg atc cgg 1225 Gly Asn Lys Wing Gln Asp Tyr Leu Cys Thr Leu Wing Ser Arg He Arg 350 355 360 agg cta gac gag agg gec cag age aga gec aag aaa gca ggc acg ctg 1273 Arg Leu Asp Glu Arg Ala Gln Ser Arg Ala Lys Lys Ala Gly Thr Leu 365 370 375 380 cct ttc age tgg gta tat ggt agg gaa gtc ca ctg tga aatcggaaac 1322 Pro Phe Ser Trp Val Tyr Gly Arg Glu Val Gln Leu 385 390 cca tgegac tgcttgagtt ggagcatagt etatcatgea ccctatgacg catcgc cga 1382 caagacctgg tgtgtcgcgt gacatagttg ttcaggtttt gaccaaatgg tctgggagca 1442 tttgttttgc cttgtgccgt ctcatagagc gttaggatag tgtacgtctg tgttctagct 1502 ttgttttgtc tgetgctttg atgtaacttg tggccatgag gctggacatg gagtgaacat 1562 gttgtaeatt gtcgctggcg gtatgtttcg gtatgttatt tcagttgctt -gagatctgtt 1622 aattttttgc gcagctatgg aggtcgttct gttctggtca aaaaaaaaaa aaaaaaaaaa 16B2 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aa 1714 210 >; 9 < 211 > 392 < 212 > PET c213 > Zea nays < 400 > 9 Met Ala Leu Arg Leu His Asp Val Ala Leu Cys Leu Ser Pro Pro Leu 1 5 10 15 Ala Ala Arg Arg Arg Ser Gly Gly Ser Phe Val Ala Ala Ala Ser Met 20 25 30 Thr Ser Ala Ala Val Ser Thr Arg Val Glu Asn Lys Lys Pro Phe Wing 35 40 45 Pro Pro Arg Giu Val Kis Val Gln Val Thr His Ser Met Pro Ser Ser 50 5 = > 60 Lys Xle Glu He Phe Lys Ser Leu Asp Asp Trp Wing Arg Asp Asn He 65 70 75 80 Leu Thr Eis Le. Lye Pro Val Glu Lys Cys Trp Gln Pro Gln Asp Phe 85 90 95 Leu Pro Asp Pro Wing Ser Glu Gly Phe Hie Aep Glu Val Lys Glu Leu 100 105 110 Arg Glu Arg Ala Lys Glu He Pro Asp Asp Tyr Phe Val Cys Leu Val 115 * 120 125 Gly Asp Met He Thr Glu Glu Ala Leu Pro Thr Tyr Gin Thr Met Leu 130 135 140 Asn Thr Leu Asp Gly Val Arg Asp Glu Thr Gly Ala Ser Pro Thr Ala 145 150 155 160 Trp Wing Val Trp Thr Arg Wing Trp Thr Wing Glu Glu Asn Arg His Gly 165 170 175 Asp Leu Leu Asn Lys Tyr Met Tyr Leu Thr Gly Arg Val Asp He Arg 180 1B5 190 Gin lie Glu Lys Thr He Gln Tyr Leu He Gly Ser Gly Met Asp Pro 195 200 205 Arg Thr Glu Asn Asn Pro Tyr Leu Gly Phe Val Tyr Thr Ser Phe Gln 210 215 220 Glu Arg Ala Thr Phe He Ser His Gly Asn Thr Ala Arg His Ala Lye 225 230 235 240 Asp Phe Gly Asp Leu "Lys Leu Wing Gln He Cys Gly He He Wing Ser 245 250 255 Asp Glu Lys Arg His Glu Thr Wing Tyr Thr Lys He Val Glu Lys Leu 260 265 .270 Phe Glu He Asp Pro Asp Gly Thr Val Val Wing Leu Wing Asp Met Met 275 280 2B5 Lye Lye Lys He Ser Met Pro Wing His Leu Met Phe Asp Gly Gln Asp 290 295 300 Asp Lys Leu Phe Glu His Phe Ser Met Val Wing Gln Arg Leu Gly Val 305 310 315 320 Tyr Tr a rg sp yr u 325 33D 335 Trp Lys Val Wing Asp Leu Thr Gly Leu Ser Gly Glu Giy Asn Lys Wing 340 345 350 Gln .Asp Tyr Leu cys Thr Leu Wing Ser. Arg He Arg Arg Leu Asp Glu 35? 36D 365 Arg Ala Gin Ser Arg Ala Lys Lys Ala Gly Thr Leu Pro Phe Ser Trp 370 375 380 Val Tyr Gly Arg Glu Val Gln Leu 3B5 390 < 210 > 10 «211 > 1709 < 212 > DNA < 21 Zea mays < 220 > < 221 > CDS < 222 > (102.}. .. (1280): 400> 10 cggcacgagc acacacaagg gaaggggaca accacaagcg cctactcctc cgtcctccgc 60 gtcgagatct ttgccgaggc ggtgaccgtc gagggatcgc c atg gcg ttg agg gcg 116 Met Ala Leu Arg Ala 1 5 tcc ccc gtg teg cat ggc acc gcg gca gcg ceg ctg ceg cet ttc gcg 164 Be PTD Val Ser Kis Gly Thr Wing Wing Pro Pro Leu Pro Pro Wing 10 15 20 cgg agg aag atg gcc cgt ggg gtg gtg gtg gcc atg gcg tcc acc atc 212 Arg Arg Lye Met Wing Arg Gly Val Val Val Wing Met Wing Being Thr He 25 30 35 aac agg gtc aaa act gtc aaa gaa ccc tat acc cct cea cga gag gta 260 Asn Arg Val Lys Thr Val Lys Glu Pro Tyr Thr Pro Pro Arg Glu Val 40 45 50 cat cgc ca att acc catta ce cct caa aag cgg gag att ttc 308 Kie Arg Gln He Thr His Ser Leu Pro Pro Gln Lys Arg Glu He Phe 55 60 65 at t he ctact cct tgg gcc aag gat aac cta a cta c a ta cta g aag 356 Asp Ser Leu Gln Pro Trp Wing Lys Asp Asn Leu Leu Asn Leu Leu Lys 70 75 B0 85 cea gtt gaa aag tea tgg cag cea cag gac ttc cta cea gag cct tct 404 Pro Val Glu Lys Ser Trp Gln Pro Gln Asp Phe Leu Pro Glu Pro Ser 90 95 100 tct gat ggg ttt tat gat gaa gtt aaa gaa ctg agg gag cgg gca aat 452 Be Asp Gly Phe Tyr Asp Glu Val Lys Giu Leu Arg Glu Arg Ala Asn 105 110 115 gaa ata cct gat gaa tac ttt gtt tgc 'tta gtt ggt gat atg gtt act 500 Glu He Pro Aep Glu Tyr Phe Val Cys Leu Val Gly Asp Met Val Thr 120 125 130 gag gac gcc tta cct here tac caca atg ctt aac act ctt gat gga 546 Glu Glu Wing Leu Pro Thr Tyr Gln Thr Met Leu Asn Thr Leu Asp Giy 135 140 145 gtc cgg gat gaa act ggt gca agt tea acc aeg tgg gcg gtt tgg ac 596 Val Arg Asp Glu Thr Gly Wing Being Thr Thr Trp Wing Val Trp Thr 150 155 160 165 agg gea tgg here gct gaa gag aac aga cat ggt gac etc ctt aac aag 644 Arg Ala Trp Thr Ala Glu Glu Asn Arg His Gly Asp Leu Leu Aen Lye 170 175 180 tac atg tac ctt act gga cgg gtt gac atg aaa ca att gag aag acc € 92 Tyr Met Tyr Leu Thr Gly Arg Val Asp Met Lys Gl n lie Glu Lys Thr 185 190 195 ata ca tat ctg att ggt tcc gga atg gat cet gga act gaac aac aac 740 He Gln Tyr Leu He Gly Ser Gly Met Aep Pro Gly Thr Glu Asn Aen 200 205 210 ccc tac ttg ggt ttc etc tac here tea ttc caa gaa agg gca ac ttt 786 Pro Tyr Leu Gly Phe Leu Tyr Thr Ser Phe Gln Glu Arg Wing Thr Phe 215 220 225 gtg teg cat ggg aat act gca agg cat gcc aag gag tat ggt gat etc B36 Val Ser His Gly Aen Thr Ala Arg His Ala Lys Glu Tyr Gly Asp Leu 230 235 240 245 aag ctg gcc cag ata tgt ggc acg ata gca gcc gat gag aag cgc falls 884 Lye Leu Wing Gln He Cys Gly Thr He Wing Wing Asp Glu Lye Arg His 250 255 260 gaa here gcc tac acc aag ata gtc gag aag cte ttc gag atg gac cct 932 Glu Thr Wing Tyr Thr Lys He Val Glu LyB Leu Phe Glu Met Asp Pro 265 270 275 gat tac ac gtg ctt gcg ttt gct gac atg atg agg aag .aag atc acg 980 Asp Tyr Thr Val Leu Wing Phe Wing Asp Met Met Arg Lys He Thr 280 2B5 290 • atg cea gcc cat etc atg tac gac ggt aag gac gac aac ctg ttc gag 1028 Met Pro Ala His Leu Met Tyr Asp Gly Lys Asp Asp Asn Leu Phe Glu 295 300 305 fall ttc age gcg gtg gcg cag agg ctg ggc gtc tac acc gcc aaa gac 1076 His Phe Ser Ala Ala Ala Gln Arg Leu Gly Val Tyr Thr Ala Lys Asp 310 315 320 325 tac acc gac atc etc gag ttc ctg gtc cag agg tgg aaa gtc gcg gag 1124 Tyr Wing Asp lie Leu Glu Phe Leu Val Gln Arg Trp Lys Val Wing Glu 330 335 340 etc here ggg ctg tct gga gaa gcg aga age gcg cag gac ttt gtc tgt 1172 Leu Thr Giy Leu Ser Giy Gl u Gly Arg Ser Ala Gln Asp Phe Val Cys 345 35D 355 acc ttg gcg ceg agg atc agg cgg ctg gat gat aga gct ca gcg agg 1220 Thr Leu Ala Pro Arg He Arg Arg Leu Asp Asp Arg Ala Gln Ala Arg 360 365 370 gcg aag caca gca ceg gtt att cct ttc agt; sg gtt tat gac cgc aag 1266 Wing Lye Gln Wing Pro Val He Pro Phe Ser Trp Val Tyr Asp Arg Lys 375 3B0 385 gtg cag ctt taa tcaagaaege taggcaatgt gggeatttae tacgtatatc 1320 Val Gln Leu 390 attttcagtc ctggggttct ctataagaaa cagtctctag gttatctagc agggtagaat 1380 gtggauctea tcaactactc ctcggtgcaa agtagtgcaa agtacgctat ctgttgttac 1440 cgtgcaagct gcagagtttg gattactatg tgggcctggt ggtggagagg aattctgtgg 1500 ggtgcctgca gccagttatg agtggcagct ecatcgcaac tgagttgttg tattgaatat 1560 gttacag ac ctatagtaac cgaaagtsat aatatgaag ttgtatatcg acaagcttgc 1620 tttggtgatt atgagaatc tgaagtaata atatggagtt tgcataaaaa aaaaaaaaaa 1680 aaaaaaaaaa aaaaaaaaaa aaaaaaaaa 1709 < 210 > 11 < 211 > 392 < 212 > PRT < 213 > Zea maye < 400 > 11 Met Ala Leu Arg Ala Ser Pro Val Ser His Gly Thr Ala Ala Ala Pro 1 5 10 15 Leu 'Pro Pro Phe Ala Arg Arg Lys Met Ala Arg Gly Val Val Val Ala 20 25 30 Met Ala Ser Thr He Asn Arg Val Lye Thr Val Lye Glu pro Tyr Thr 25 40 45 Pro Pro Arg Glu Val His Arg Gln He Thr His Ser Leu Pro Pro Gln 50 55 60 Lys Arg Glu He Phe Asp Ser Leu Gln Pro Trp Wing Lys Asp Asn Leu 55 70 75 80 Leu Asn Leu Leu Lys Pro Val Glu Lys Ser Trp Gln Pro Gln Asp Phe 85 90 95 Leu Pro Glu Pro Ser Asp Gly Phe Tyr Asp Glu Val Lys Glu Leu 100 IOS 110 Arg Glu Arg Wing Asn Glu He Pro Asp Glu Tyr Phe Val Cys Leu Val 115 120 125 Gly Aep Met Val Thr Glu Glu Ala Leu Pro Thr Tyr Gln Thr Met Leu 13C 135 140 Asn Thr Leu Asp Gly Val Arg Asp Glu Thr Gly Ala Ser Ser Thr Thr 145 150 15S 160 Trp Wing Val Trp Thr Arg Wing Trp Thr Wing Glu Glu Aen Arg His Gly 165 170 175 Asp Leu Leu Asn Lys Tyr Met Tyr Leu Thr Gly Arg Val Asp Met Lys 180 1B5 190 Gln He Glu Lys Thr He Gln Tyr Leu He Gly Ser siy Met Asp Pro 195 200 205 Gly Thr Glu Asn Asn Pro Tyr Leu Gly Phe Leu Tyr Thr Ser Phe Gln 210 215 220 Glu Arg Ala Thr Phe Val Ser His Gly Asn Thr Ala Arg Kis Ala Lys 225 230 235 240 Glu Tyr Giy Asp Leu Lys Leu Wing Gln He Cys Gly Thr He Wing Wing 245 250 255 Asp Giu Lys Arg His Giu Thr Wing Tyr Thr Lys He Val Glu Lys Leu 260 265 270 Phe Glu Met Asp Pro Asp Tyr Thr Val Leu Wing Phe Wing Asp Met Met 275 280 285 Arg Lys Lys He Thr Met Pro Wing Hie Leu Met Tyr Asp Gly Lye Aep 290 295 300 Asp Asn Leu Phe Glu His Phe Be Wing Val Wing Gln Arg Leu Gly Val 305 310 315 320 Tyr Thr Wing Lys Asp Tyr Wing Asp lie Leu Glu Phe Leu Val Gln Arg 325 330 335 Trp Lys Val Wing Glu Leu Thr Gly Leu Ser Gly Glu Gly Arg Ser Wing 340 345 350 Gln Aep Phe Val Cys Thr Leu Wing Pro Arg He Arg Arg Leu Asp A3p 355 360 365 Arg Wing Gln Wing Arg Wing Lys Gln Wing Pro Val He Pro Phe Ser Trp 370 375 380 Val Tyr Asp Arg Ly = Val Gln Leu 3B5 390 < 210 > 12 < 211 > 18 212 > DNA < 213 > ARTIFICIAL SEQUENCE r ^ 22 D > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE < 400 > 12 aggacgctac cgtaggaa 18 < 210 = > 13 < 2ll > 17 «212 > DNA c213 > ARTIFICIAL SEQUENCE < 220 > c223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO c4DD > 13 gcgatggcac tgca ta 17 < 210 > 14 «.211 > 21 < 212 > DNA «.213 > ARTIFICIAL SEQUENCE < 220 > c223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO < 400 > 14 cttgagagaa gaaccacact c 21 < 210 > 15 < 211 > 21 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE < 220 > «223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OUGONUCLEOTTDO < 400 > 15 ctagacatat cgagcatgct g 21 < 21D > 16 c211 > 22 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE < 220 > «.223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: OLIGONUCLEOTIDO SINTE? CO c400 > 16 aggcgctgac ggtggcgacg ct 22 < 210 > 17 < 211 > 20 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE c220 > < 223 > DESCRIPTION OF THE SEQUENCE ARTTFIC L: SYNTHETIC OLIGONUCLEOTIDE c4D0 > 17 gtgttggcga ggcacgtgag 20 < 210 > 18 < 2ll > 46 < 212: > DNA < 2 3 > ARTIFICIAL SEQUENCE 220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OUGONUCLEOT < 400 > 18 acctcccgtc gcaccccggt ggtgatcagc catggtaggc tagcag 46 < 210 > 19 < 211 > 1714 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE -22D < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOT < 4O0 > 19 tctagaa tg tatgtatgtc aaagatctta tcgggataag agatatgata aagatcttaa 60 cggaatcaga gccaggtttg taaaaataga gttggactcg tgtacaactt ggtctctggc 120 ttagctccgt catgaattta gtaaccgact cgatatgtac cgtggaaecc ctagggcatg 180 agccatagga tcatcatatc ccaacaaatc caaacatgca caccacacat cgaagatcca 240 tattaagaa gggttatcta ctttacaatt tcagagtaac caatagagcc aaactc tag 300 cacaggggag cttcatatca gatggagcca ttgaattgat ataaaaagct gaagttctaa 360 aaasttt aa gtgctggaac ttcaaagccg ctaactagtg aagcaccgaa gccttccgtg 420 gagagataca tacacgacac gttagggacg taaaatgacg gaattataca gctacctcta -480 tatgtgacac ttatgtaata gaaaagacag aatccatatg aagatgtata atggatcaat 540 catataaata gataaacaat tgaggtgttt ggtttgatga atcactctat ccaaaataaa 600 gtggtgcatc atgggtttat tcctcaaatt tggtggcatg actacat acatattagt 660 cc actaageaac taactttgag gaatgaggtg atgatgaatt aactcactcc attccacaaa 720 ccaaacaaaa atttgaggag tgagaagatg attgactatc tcattcctca aaccaaacac 780 ctcaaatata tctgctatcg ggattggcat tcctgtatcc ctacgcccgt gtaccccctg 840 tttagagaac ctcccaa agg tataagatgg cgaagattat tgttgtcttg tctttcatea 900 tatatcgagt ctttccctag gatattatta ttggcaatga gcattacacg gttaatcgat 960 tgagagaaca tgcatctcac cttcagcaaa taattacgat aatccatatt ttaegcttcg 1020 tgagtttcga taacttctca tatacaaatt tgttttctgg acaccctacc attcatcctc 1080 ttcggagaag agaggaagtg tectcaattt aaatatgttg gttetteaca tcatgctgta 1140 aaatctcaac aggtaccaag cacattgttt ccacaaatta tattttagtc acaataaatc 1200 Jtatattatta ttaatatact aaaactatac tgacgctcag atgcttttac tagttcttgc 1260 tgtaggtcta tagtatgtga cgtggaccag aaaatagtga gacacggaag acaaaagaag 1320 taaaagaggc ccggactacg gcccacatga gattcggccc cgccacctcc ggcaaccagc 1380 ggccgatcca acggcagtgc gcgcacacac acaacctcgt ata atcgcc gcgcggaagc 1440 ggcgcgaceg aggaagcett gtcctcgaca ccccctacac aggtgtcgcg ctgcccccga 1500 caegagtcet gcatgcgtcc eacgcggccg cgccagatcc cgcctccgcg cgttgccacg 1560 ccctctataa cacccagct ctccctcgcc ctcatctacc tcactcgtag tcgtagctca 1620 agcatcagcg gcagcggcag cggcaggagc tctgggcagc gtgcgcacgt ggggtaccta 1680 gctcgctctg ctagcctacc atgg taegtg gcat 1714 < 210 > 20 < 211 > 32 C213 > JARTIFICIAL SEQUENCE 220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OUGONUCLEOTIDE < 400 > 20 cttatgtaat agaaaagaca ggatccatat gg 32 < 210 > 21 < 211 > 33 c2X2 > DNA < 213 > ARTIFICIAL SEQUENCE < 220 > c223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO < 400 > 21 gaggagtgag gatcctgatt gactatctca ttc 33 < 210 > 22 c211 > 33 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE c220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «: 400 > 22 tctggacacc ctaccattgg atcctcttcg gag 33 «, 210 > 23 c211_. 32 «212-. DNA c213 > ARTIFICIAL SEQUENCE < 220 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOT1DO < 223 > < 400 > 23 agagttggat ccgtgtacaa cttggtctct gg 32 < 210 > 24 < 211 > 37 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE «220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «40D > 24 gccgctgatg ctcgagctac gactacgagt gaggtag 37 < 210 > 25 < 211 > 32 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE «-220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE < 40D > 25 Btgcgggact cgagtcgggg gcagcgcgae ac 3 < 210 > 26 * 211 > 32 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE < 220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE «: 40D > 26 gtggcggggc cgaatctcga gtgggccgta gt 3 c210 > 27 < 211 > 33 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE < 220 > c223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE < 400 > 27 gccacgtgcc atggtaggct agcagagcga gct 33 < 210 > 28 < 211 > 24 «212 > DNA < 213 > ARTIFICIAL SEQUENCE < 220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE • < 400 > 28 aacacacace catggatatc acag 24 < 210 > 29 «.211 > 19 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE < 22 D > < : 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE < 40D > 29 ggtctgactt acgggtgtc 19 < 210 > 30 < 211 > 25 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE 220. < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «.400 > 30 ctctcccgtc ctcgagaaac cctcc 2 < 210 > 31 < 211 > 25 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE c220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE < 400 > 31 cttggcagcc atggctcgat ggttc 25 c210 > 32 < 211 > 30 c212 > DNA < 213 ARTIFICIAL SEQUENCE < 220 > -.223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE < 400 > 32 atgg gagcg ccagaatcgt tgtcctcctc 30 < 210 > 33 < 211 > 30 < 12 > DNA < 213- «ARTIFICIAL SEQUENCE < 220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO < 400 > 33 catcctggcg gtcaccatcc tcaggagcgt 30 < 210 > 34 c211 > 30 < 212 > DNA c213 > ARTIFICIAL SEQUENCE c220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: OLIGONUCLEOTJDO SYNTHETIC c400 > 34 atagggaatt ctctgttttt ctaaaaaaaa 30 210 > 35 < 211 > 30 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE < 220 > '< 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: OLIGONUCLEO TTDO SINTÉTICO < 400 > 35 gctcaccatg gtgtagtgtc tgtcactgtg 30 c210 > 36 < 211 > 35 «212 > DNA < 213 > ARTIFICIAL SEQUENCE 220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE < 400 > 36 ggggga cca agcttgagga gacaggagat aaaagt 36 < 210 > 37 < 211 > 39 < 212 > DNA «213 > ARTIFICIAL SEQUENCE c220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO < 400 > 37 gggctgcagc tcgagggtgt agtgtctgtc actgtgata 3 $ < 210 > 3B < 211 > 1108 < 212 > DNA < 213 > ARTIFICIAL SEQUENCE «220 > < 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE < 4O0 > 3B atccatatga agatgtataa tggatcaatc atataaatag ataaacaatt gaggtgtttg 60 gtttgatgaa tcaetctatc caaaataaag tggtgeatca tgggtttatt cctcaaattt 120 ctacattcca ggtggcatga catattagta ctaagcaact aactttgagg aatgaggtga 180 tgatgaatta actcacteca ttccacaaac caaacaaaaa tttgaggagt gagaagatga 240 ttgactatct cattcctcaa accaaacacc tcaaatatat ctgctatcgg gattggcatt 300 cct tatccc tacgcccgtg taccccctgt ttagagaacc tcccaaaggt ataagatggc 360 gaagattatt gttgtcttgt ctttcatcat atategagtc tttccctagg atattattat 420 tggcaatgag cattacacgg ttaatcgatt gagagaacat gcatctcacc ttcagcaaat 480 aattacgata atccatattt tacgcttcgt aaettetcat gagtttcga atacaaattt 540 caccctacca gttttctgga ttcatcctct tcggagaaga gaggaagtgt cetcaattta 600 aatatgttgt catgctgtag ttcttcacaa aatetcaaca ggtaccaagc acattgtttc 660 attttagtca cacaaattat caataaatct atattattat taatatact aaactatact 720 gacgctcaga tgcttttact agttcttgct agtatgtgat gtaggtctac gtggaccaga 780 aa agtgag cacggaaga ßaaaagaagt aaaagaggcc cggactacgg cccacatgag 840 attcggcccc gccacctccg gcaaceagcg gccgatccaa cggcagtgcg cgcacacaca ca 900 cctcgta tatatcgecg egeggaagcg gcgcgaccga ggaagccttg tcctcgacac 960 cccctacaca ggtgtegcgc tgcccccgac acgagteccg eatgcgtccc ac cggccgc 1020 gecagatccc gcctccgcgc gttgccacgc cetetataaa cacccagctc tccctcgccc 1080 tcatctacct cactcgtagt cgtagctc 1108 < 210 > 39 < 211 > 871 «212 > DNA «213 > Zea mays < 400 > 39 tgattgacta tctcattcct caaaccaaac acctcaaata tatctgctat cgg attggc 6C attcctgtat ccctacgccc gtgtaeccce tgtttagaga acctcccaaa ggtataagat 120 ggcgaagatt attgttgtct tg ctttcat catatatega gtctttccct aggatattat 180 tattggcaat gagc raca cggttaatcg attgagagaa catgcatctc accttcagca 240 aataattacg ataatccata ttttacgctt cgtaacttct catgagtttc gatataeaas 300 tttgttt ct ggacaccc ccattcatcc tcttcggaga agagaggaag tgtcctcaat 360 ttaaatatgt tgtcatgctg tagttcttca caaaatctca agcaca acaggtacca tgt 420 ttccacaaat tatattttag tcacaataaa tetatattat tattaatata ctaaaactat 480 actgacgctc agatgctttt actagttctt gctagtatgt gatgtaggtc tacgtggacc 540 agaaaata t ag eacgga agacaaaaga agtaaaagag gcccggacta cggcecacat 600 g attcggC cccgccacct ccggcaacca gcggccgatc caacsgcagt gcgcgeacac 660 acacaacctc gtatatatcg ccgcgcggaa gcggcgcgac cgaggáagcc ttgtcctcga 720 caceeectac acaggtgtcg cgctgccccc gacaegagtc ccgcatgcgt cccacgcggc 7B0 egcgcc gat cccgcctccg cgcgttgcca cgccctctat aaacacccag ctctccctcg 840 ccctcatcta cctcactcgt agtcgtagct c 871 c210 >; 40 c211 > 545 < 212 > DNA «213 > Zea mays < 400 > 40 atcctcttcg gagaagagag gaagtgtcct caatttaaat atgttgtcat gctgtagttc 60 ttcacaaaat ctcaacaggt accaagcaca ttgtttccac aaattatatt ttagtcacaa 120 ttattattaa taaatctata tatactaaaa ctatactgac gctcagatgc ttttactagt 180 atgtgatgta tcttgctagt ggtctacgtg gaccagaaaa tagtgagaca cggaagacaa 240 aagaag aaa agaggcccgg actacggccc catgagatt cggccccgcc acctccggca 300 accagcggcc gatecaaegg cagtgcgcgc acacacacaá cctcgtatat atcgccgcgc 360 ggaagcggcg cgaccgagga agccttgtcc tcgacacccc ctacacaggt gtcgcgctgc 420 ccccgacacg agtcccgcat gcgtcccaeg cggccgcgcc agatcccgcc tccgcgcgtt 480 gccacgccct etataaacac ccagctctcc ctcgccctca tctacctcac tcgtag cgt 540 agetc 545 «210 > 41 < 211 > 952 «212 > DNA c213 > Zea mays «400 :. 41 tgattgacta tctcattcct caaaccaaac acctcaaata tatctgctat cgggattggc 60 attcctgtat ccctacgccc gtgtaccccc tgtttagaga acctcccaaa ggtataagat 120 ggcgaagatt attgttgtct tgtctttcat catatatega gtctttcect aggatattat 180 tattggcaat gageattaca cggttaateg attgagagaa catgcatctc "accttcagca 240 aataattacg ataatccata ttttacgctt cgtaacttct catgagtttc tacaaa ga 300 tttgttttct ggacacccta ccattcatcc tcttcggaga agagaggaag tgtcctcaat 360 ttaaatatgt tgtcatgctg tagttcttca caaaatctca acaggtacca agcacattgt 420 ttccacaaat tatattttag tcacaataaa tetatattat tattaatata ctaaaactat 480 actgacgctc agatgctttt actagttctt gctagtatgt gatgtaggtc tacgtggacc 540 agaaaatagt gagacacgga agacaaaaga agtaaaagag gcccggacta cggcccacat 600 gagattcggc cccgccacct ccggcaacca gcggccgatc caacggcagt gcgcgcacac 660 acacaacctc gtatatatcg ccgcgcggaa gcggcgcgac cgaggaagce ttgtcctcga 720 caccecctac acaggtgtcg cgctgccccc gacaegagtc ccgcatgcgt cccacgeggc 780 cgcgccagat cccgcctccg cgcgttgcca cgccctctat aaacacccag ctctccctcg 840 ccctcatcta cctcac tcgt agtcgtagct caageatcag cggcagcggc agcggcagga 900 gctctgggca gcgtgcgcac gtggggtacc tagetcgctc tgctagccta ce 952 «210 > 42 < 211 > 1403 «212 > DNA «213 > Zea maye < 400 > 42 cgtgtacaac ttggtctctg gcttagctcc gtcatgaatt tagtaacega etegatatgt 60 accgtggaac ccctagggca tgagccatag gatcatcata tccaaacatg caccaacaaa 120 ategaagate tccaccacac catattaaga aggggttatc tactttacaa tttcagagta 1B0 accaa agag ccaaactcat agcacagggg agetteatat cagatggagc cattgaattg 240 atataaaaag ctgaagttct aaaaagtttt aagtgctgga acttcaaagc cgctaactag 300 tgaagcaccg aagccttccg tggagagata catacacgac egtaaaatga acgttaggga 360 cggaattata cagetacctc tatatgtgac acttatgtaa tagaaaagac agaatecata 420 taatggatca tgaagatgta tagataaaca atcatataaa attgaggtgt ttggtttgat 4B0 gaatcactct atccaaaata aagtggtgca attcctcaaa tcatgggttt tttggtggca 540 tgactacatt ccacatatta gtactaagea actaactttg aggaatgagg tgatgatgaa 600 ttaactcact ccattccaca aaatttgagg aaccaaacaa agtgagaaga tgattgacta 660 tctcattcct caaaccaaac acctcaaata tatctgctat cgggattggc attcctgtat 720 ccctacgccc gtgtaccccc tgtttagaga acctcccaaa ggtataagat ggcgaagatt 780 attgttgtct tgtctttcat catatatega gtctttccet aggatattat tattggcaat 840 gageattaca cggttaa tcg attgagagaa catgcatctc accttcagca aataattacg 900 ataatccata ttttacgctt cgtaacttct catgagtttc gatatacaaa tttgttttct 960 ggacacccta ccattcatcc tsttcggaga agagaggaag tgtcctcaat ttaaatatgt 1020 tagttcttca tgtcatgctg caaaatctca acaggtacca agcacattgt ttccacaaat 1OB0 tatattttag tcacaataaa tetatattat tattaatata ctaaaáctat actgacgctc 1140 agatgctttt actagttctt getagtatgt gatgtaggtc tacgtggacc agaaaatagt 1200 agacaaaaga gagacacgga agtaaaagag gcccggacta cggcccacat gagattcggc 1260 ccggcaacca cccgccacct gcggccgatc caacggcagt gcgcgcacac acacaacctc 1320 gtatatatcg ccgcgcggaa gcggcgcgac cgaggaagcc ttgtcctcga caccccctac 1380 acaggtgtcg cgctgccccc gac '1403 «21D > 43 «211 > 990 «212 > DNA «21 > Zea mays «400 > "43 atccatatga agatgtataa tggatcaatc atataaatag ataaacaatt gaggtgtttg 60 gtttgatgaa tcactcfcatc caaaataaag tggtgeatca tgggtttatt cctcaaattt 120 gcatga gg catattagta ctacattcca ctaagcaact aactttgagg aatgaggtga 180 tgatgaatta actcactcca ttccacaaac caaacaaaaa tttgaggagt gagaagatga 240 ttgactatct cattcctcaa accaaacacc tcaaatatat ctgctatcgg gattggcatt 300 cc tatccc tacgcccgtg taccccctgt ttagagaacc tcccaaaggt ataagatggc 360 gaasat att gttgtcttgt ctttcatcat atategagtc tttccctagg- atattattat 420 tggcaatgag cat acaegg ttaategatt gagagaacat gcatctcacc ttcagcaaat 48 aattaegata atecatattt tacgcttcgt aaettetcat gagtttcgat atacaaattt 540 caecctacca gttttctgga ttcatcctct tcggagaaga gaggaagtgt cetcaattta 600 aatatg tgt catgctgtag ttetteacaa aatetcaaca ggtaccaagc acattgtttc 660 attttagtca cacaaattat caataaatct atattattat taatatacta aaactatact 720 gaege shits tgcttttact agttcttgct agtatgtgat gtaggtctac gtggaccaga 780 aaatagtgag acacggaaga caaaagaagt aaaagaggcc cggactacgg cccacatgag 840 attcggcccc gccacct ccg gcaaccagcg gccgatccaa cggeag gcg cgcacacaca 900 caacctcgta tatategecg cgcggaagcg gcgcgaccga ggaagccttg tcctcgacac 960 cccctacaca ggtgtcgcgc tgcccccgac 990 «210 >; 44 «211 > 753 «212 > DNA «213 > Zea mays < 400 > 44 tgattgacta tctcattcct caaaccaaac acctcaaata tatetgetat cgggattggc 60 attcctgtat ecctacgccc. gtgtaccecc tgtttagaga acctcccaaa ggtataagat 120 ggcgaagatt attgttgtct tgtctttcat catatatega gtctttccct aggatattat 18C tattggcaat cagcattaca cggttaatcg attgagagaa catgcatctc accttcagca 240 aataattacg ataatccata ttttacgctt cgtaacttct catgagtttc gatatacaaa 300 tttgttttct ggacacccta ccattcatce tcttcggaga agagaggaag tgtcctcaat 360 ttaaatatgt tgtcatgctg eaaaatctca tagttcttca acaggtacca agcacattgt 42c ttccacaaat tatattttag tcacaataaa tetatattat tattaatata ctaaaactat 480 acgaegetc agatgctttt actagttctt gctagtatgt gatgtaggtc tacgtggacc 540 agaaaatagt gagacacgga agacaaaaga agtaaaagag gcccggacta cggcccacat 600 gagattegsc cccgccacct ccggcaacca gcggccgatc caacggcagt gcgcgcacac 660 acacaacctc gtatatatcg ccgcgcggaa gcggcgcgac cgaggaagcc ttgtcctcga 720 753 gac caccccctac acaggtgtcg cgccgccecc «210 > 45 < 211 > 427 «212 > ADÑ «13 > Zea mays «400 > 45 atcctcttcg gagaagagag ga & gtgtcct eaatttaaat atgttgtcat gctgtagttc 60 ttcacaaaat ctcaacaggt accaagcaca ttgtttccac aaattatatt ttagtcacaa 120 ttattattaa taaatctata tatactaaaa ctatactgac gc cagatgc ttttactagt 1B0 tcttgctagt atgtgatgta ggtctacgtg tagtgagaca gaccagaaaa cggaagacaa 240 aagaag aaa agaggcccgg actacggccc acatgagatt cggccccgcc acctccggca 300 accagcggcc gatecaaegg cagtgcgcgc acacacacaa cctcgtatat atcgccgcgc 360 ggaagcggcg cg ccgagga agccttgtcc tcgacacccc ctacacaggt gtcgcgctgc 420 ccccgac 427 «210 > 46 «211 > 1248 «212 > DNA «213 > .Zea mays «400 > 4-6 cgtgtacaac ttggtctctg gcttagctcc gtcatgaatt tagtaacega etegatatgt 60 accgtggaac ccctagggca tgagccatag gatcatcata tccaaacatg caccaacaaa 120 ategaagate tccaecacac catattaaga aggggttatc tactttacaa tttcagagta 1B0 accaatagag ccaaactcat agcacagggg agcttcatat cagatggagc cattgaattg 240 atataaaaag ctgaagttct aaaaagtttt aagtgctgga acttcaaagc cgctaactag 300 tgaagcaecg aagccttccg tggagagata catacácgac cgtaaaatga acgttaggga 360 cggaattata cagctacctc tatatgtgac acttatgtaa tagaaaagac agaatecata 420 gta tgaaga taatggatca atcatataaa tagataaaca attgaggtgt ttggtttgat 4B0 gaatcactct atccaaaata aagtggtgca attcctcaaa tcatgggttt tttggtggca 540 tg ctacatt ccacatatta gtactaagea actaactttg aggaatgagg tgatgatgaa 600 ccattccaca etaactcaet aaatttgagg aaccaaacaa agtgagaaga tgattgacta 660 tctcattcct caaaccaaac acctcaaata tatctgctat cgggattggc attcctgtat 720 ccctacgccc gtgtaccccc tgtttagaga acctcccaaa ggtataagat ggcgaagatt 780 attgttgtct tgtctttcat catatatega gtctttccct aggatattat tattggcaat 840 gageattaca cggtt aatcg attgagagaa catgcatctc accttcagca aataattacg 900 ataatccata ttttacgctt cgtaacttct catgagtttc gatatacaaa tttgttttct 960 ggacacccta ccattcßtcc tc tcggaga agagaggaag tgtcctcaat ttaaatatgt 102C tgtcatgctg tagttcttca acaggtacca caaaatctca ttccacaaa agcacattgt 1080 tatattttag tcacaataaa tetatattat tattaatata ctaaaactat actgacgctc 1140 agatgctttt actagttctt gctagtatgt gatgtaggtc tacgtggacc agaaaatagt 1200 agacaaaaga gagacacgga agtaaaagag gcccggacta cggcccac 124B «210 > 47 «211 > 835 «212 > DNA «213 > Zea maye < 4O0 > 47 atccatatga agatgtataa tggatcaatc atataaatag ataaacaatt gaggtgtttg 60 g ttgatgaa tcactctatc caaaa.taaag tggtgeatca tgggtttatt cctcaaattt 120 ctacattcca ggtggcatga catattagta ctaagcaact aactttgagg aatgaggtga 180 tgatgaatta aetcactcca ttccacaaac caaacaaaaa tttgaggagt gagaagatga 240 ttgactatct cattcctcaa accaaacacc tcaaatatat ctgctatcgg gattggcatt 300 cctgtatccc tacgcccgtg taccccctgt ttagagaacc tcccaaaggt ataagatggc 360 gaagattatt gttgtcttgt ctttcatcat atategagtc tttccctagg atattattat 420 tggcaatgag cattacaegg ttaategatt gagagaacat gcatctcacc ttcagcaaat 4B0 aattaegata atecatattt tacgcttcgt aaettetcat gagtttcgat atacaaattt 540 caccctaeca gttttctgga ttcatcctct tcggagaaga saggaagtgt cetcaa ta 600 aat tgt gt catgctgtag ttetteacaa aatetcaaca ggtaccaagc acattgtttc 660 attttagtca cacaaattat caataaatct atattattat taatatacta aaactatact 720 gaegetcaga tgcttttact agttcttgct agtatgtgat gtaggtctac gtggaccaga 780 ßaatagtgag acacggaaga caaaagaagt aaaagaggcc cggactacgg cccac 835 «210? 48 «211 > 598 «213 > Zea mays «400 > 48 tgattgacta tctcattcct caaaccaaac acctcaaata tatctgctat cgggattggc 60 attcctgtat ccctacgccc gtgtaccccc tgtttagaga acctcccaaa ggtataagat 120 ggcgaagatt attgttgtct tgtctttcat catatatega gtctttccct aggatattat 180 tattggcaat gageattaca cggttaatcg attgagagaa catgcatctc accttcagca 240 aataattacg ataatccata ttttacgctt cgtaacttct catgagtttc gatatacaaa 300 tttgttttct ggacacccta ccattcatcc tcttcggaga agagaggaag tgtcctcaat 360 ttaaatatgt tgtcatgctg tagttcttca acaggtacca caaaatctca agcacattgt 420 ttccacaaat tatattttag tcacaataaa tetatattat tattaatata ctaaaactat 480 actgacgctc agatgctttt actagttctt gctagtatgt gatgtaggtc tacgtggacc 540 agaaaatagt gagacacgga agacaaaaga agtaaaagag gcccggacta cggcccac 59B «210 > 49 «211 > 272 «212 > DNA «213 > Zea snays «400 > 49 atcctcttcg gagaagagag gaagtgtcct caatttaaat atgttgtcat gctgtagttc 60 ttcacaaaat ctcaacaggt accaagcaca ttgtttccac aaattatatt ttagtcacaa 120 ttattattaa taaatctata tatactaaaa etatactgac gctcagatgc ttttactagt 180 atgtgatgta tcttgctagt ggtctacgtg gaccagaaaa tagtgagaea cggaagacaa 240 taaa AAGAA agaggcccgg actacggccc ac 272 '210 > .50 «2H: > 29 «212 > DNA «213 > ARTIFICIAL SEQUENCE «220 > «223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «400 > 50 cggggtaccg atgaccgaga aggagcggg 29 «210 > 51 «211 > 29 «212 > DNA «213 > ARTIFICIAL SEQUENCE «220s« 223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «400 > 51 ggcggtacct agaact ctt gttgtacca 2 «210 > 52 «211 > 31 «212 > DNA «213 > ARTIFICIAL SEQUENCE «220 > «223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «400 > 52 ggcctccgcc atggcgctcc gctccacgac g 31 «210 > 53 «211 > 30 «212 > DNA «213 > ARTIFICIAL SEQUENCE «220 > «223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE «400 > 53 ctccaactca agcagtcgcc atgggtttcc 30 «210 > 54 «211 > 20 «212 > DNA «213 > ARTIFICIAL SEQUENCE «220 > «223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «400 > 54 ctgcactgaa agttttggca 20 «210 > 55 «211 > . 25 «212 > DNA «213 > ARTTFICLAL SEQUENCE «220 > «223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTIDE «400 > 55 agtacagcgg ccaggcggcg tagcg 25 «210 > 56 «211 > 20 «212 > DNA «213 > ARTIFICIAL SEQUENCE «220 > «223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «400 > 56 aaggggagag agaggtgagg 20 «210? 57 «211 > -20 «212 > DNA «213 > ARTIFICIAL SEQUENCE «220 > «223 > DESCRIPTION OF THE ARTIFICIAL SEQUENCE: SYNTHETIC OLIGONUCLEOTTDO «40D > 57 tgcattgaag gtggtggtaa 20 «210 > 58 «211 > 6337 «212 > DNA «213 > Zea mays «400 > 5B gaggatccga gtcgactcta ttgactatct cattcctcca aacccaaaca cctcaaatat 60 -atctgctatc gggattggca ttcctgtatc cctacgccsg tgtaccccct gtttagagaa 120 cctcccaagg tataagatgg cgaagattat tgttgtcttg tctttcatca tatatcgagt 180 ctttccc ag -gatattatta ttggcaatga gcattacacg gttaatcgat tgagagaaca 240 tgcatctcac cttcagcaaa taattacgat aatccatatt ttacgcttcg taacttctca 300 tgagtttcga tatacaaatt tgttttctgg acaccctace attcatcctc ttcggagaag 360 agaggaagtg tcctcaattt aaatatgttg tcatgctgta gttcttcacc caatctcaac 42D aggtaccaag cacattgttt ccacaaatta tattttagtc acaataaatc tatattatta 480 ttaatatact aaaactatac tgacgctcag atgcttttac tagttcttgc tagtatgtga 540 tgtagg cta cgtggaccag aaaatagtga gacacggaag acaaaagaag taaaagaggc 600 ccggactacg geccacatga gattcggccc cgecacctcc ggcaaccagc ggccgatcoa 660 acggaagtgc gcgcacacac acaacctcgt atatatcgce gcgcggaagc ggcgcgaccg 720 aggaagcctt gftcctcgaca ccccctacac aggtgtcgcg ctgcccccga cacgagtccc 780 gcatgcgtcc cacgcggccg cgccagatcc cgcctccgcg cgttgccacg ccctctataa 840 acacccagct ctcc ctcgcc ctcatctacc tcactcgtag tcgtagctcg agaaaccctc 900 cctccc cct ccattggact gcttgctccc tgttgaccat tggggtatgc ttgctctcct 960 gttcatctcc gtgctaaacc tctgtcctct gggtgggttt ttgctgggat tttgagctaa 1020 tctgctggcc gcggtagaaa agaccgtgtc ccctgatgag ctcaagcgct cgccttagcc 1080 gcgtccttgt cecccgccat ttcttgcggt ttegctgtgt tcccgtgact cgccgggtgc 1140 gtcatcgcct gaatcttgtc tgggctctge tgacatgttc ttggctagtt gggtttatag 1200 ATTCC c ga gggttttggt tacgfcsgtsg tastaagc t gg t falls atggataaag ttgttctaag 1320 ctccgtggtt tgcttgagat cttgctgtta ttgcgtgccg tgctcacttc ttttgcaatc 1380 cgaggaatga atttgtcgtt nactcgtttt ggtggattat tagcgcgaaa aaaaactctt 1440 tttttttgtt cttttactac gaaaagcatc ttcttggatt ttgctatctt cttttactac 1500 ga aactct tgagtctagg aatttgaatt tgtgatgtcc attcttgoag tgcgctgtgc 1560 tttattggga agccaaatcc tattattttc tgcctctagg gtctgaatgg aatcagcact 1620 aatcaatcca attgagacaa atcaagttga tttctttctt taaaaata atcacagaac 16B0 taagtgcttg tgcggaatca gtactggctt ttgtttggtg gaggatcaat acttgctttt 1740 gttttggggt ggca actgtt ttgctataag attccatgtg ttcctgttga gatgaatcat 1800 atatagtata gctgcatact acaaatctgt ttttcaaatt taggttgctt tggcatgatc 1860 aatttttttt cagacagtct ttctaagtgr 'tagctcttga tttcttgttc ttctacaact 192D ggtgctgctg aatcttgacc gta ageteg aattgcagta ttctgaacca tegagecatg 1980 .aattcccccg atgacegaga aggagcggga gaagcaggag cagctcgccc gagetacegg 2040 tggcgccgcg atgcagcggt cgccggtgga gaagcctccg ttcactctgg gteagatcaa 2100 gaaggccatc ccgccacact gcttcgagcg ctcggtgctc aagtccttct cgtacgtggt 2160 ccacgacctg gt to cgccg cggcgctcct ctacttcgcg ctggccatca tacsggcgct 2220 cccaagcccg ctccgctacg ccgcctggcc gctgtactgg atcgcgcagg ggtgcgtgtg 2280 caccggcgtg tgggtcatcg cgcacgagtg cggccaccac gccttctcgg actactcgct 2340 cctggacgac gtggtcggcc tggtgctgca ctcgtcgctc atggtgccct acttctcgtg 2400 gaagtacagc caccggcgcc accactccaa cacggggtcc ctggagcgcg acgaggtgtt 2460 cgtgcccaag aagaaggagg cgctgccgtg gtaeaccccg tacgtgtaca acaacccggt 2520 cggccgggtg gtgcacatcg tggtgcagct caccctcggg tggccgctgt acctggcgac 2580 caacgcgtcg sggcggccgt accc cgctt cgcctgceac ttcgacccct acggccccat 2640 ctacaacgac cgggagcgcg cccagatctt cgtctcggac gccggcgtcg tggccgtggc 2700 gttcgggctg tacaagctgg cggcggcgtt cggggtctgg tgggtggtgc gcg gtacgc 2760 cgtgccgctg ctgatcgtga acgcgtggct ggtgctcatc acctacctgc agcacaccca 2B20 cccgtcgctc ccccactacg actegagega gtgggactgg ctgcgcggcg cgctggccac 28B0 catggaccgc gactacggca tcctcaaccg cgtgttccac aacatc cgg ac cgcacgt 2940 cgcgcaccac ctcttctcca ccatgccgca ctaccacgec atggag cca ccaaggcgat 3000 caggccc tc ctcggcsact actaccactt cgacccgacc cctgtegcca aggcgacccg 3060 ggggaatgca gcgcgaggcc tctacgtcga gcccgaggae cgcaagggcg tcttctggta 3120 caacaagaag ttctaggggg gtacctaaag aaggagtgcg tegaageaga tcgttcaaac 3180 atttggcaat aaagtttctt aagattgaat cctgttgccg gtcttgcgat gattatcata 3240 taatttctgt tgaattacgt taagcatgta ataattaaca tgtaatgcat gacgttattt 3300 atgagatsgg tttttatgat tagagtcccg caattataca tttaataege gatagaaaac 3360 aaaatatagc gcgcaaacta ggataaatta tcgcgc cg tg catctat gttactagat 3420 cg GTCGAC tetagaaage ttactagtga tgcatattct atagtgtcac ctaaatctgc 34B0 ggccgctgac caagtcaget tggcactggc cg cgtttta caacgtcgtg actgggaaaa 3540 cectggcgtt acccaactta atcgccttgc agcacatccc cct tc gctggcgtaa cca 3600 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgeagcctga tg ggega 3660 aacgttaata ggaaattgta ttttgttaat attttgttaa aattcgcgtt aaatttttgt 3720 cattttttaa taaatcagct gaaatcggca ccaataggcc aa tecetta taaatcaaaa 3780 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtec acta ttaaag 3840 aaegtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt 3900 gaaecatcac ectaatcaag ttttttgggg tcgággtgce gtaaagcact aaatcggaac 3960 cctaaaggga tgccccgatt tagagcttga cggggaaagc cggcgaacgt t? gcgagaaag 4020 aagcga gaagggaaga agg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg 4080 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggc CGTC aggtggcact 4140 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tetaaataca ttcaaatatg 4200 tgagacaata tatccgctca accctgataa atgcttcaat aatattgaaa aaggaagagt 4260 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 4320 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg gttgggtgca ctgaagatca 4380 egagtgggtt acategaact ggatetcaac ageggtaaga tccttgagag ttttegcccc 4440 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 4500 cgt ttgacg ccgggcaaga gcaactcggt cgccgcatac actattetca gaatgacttg 4560 gtcgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 4620 tgcagtgctg ccataaccat gagtgatáac aetgßggcca acttacttct gacaae gate 46B0 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 474c gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 4B00 cc tagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact • tactetaget 4B60 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgegc 4920 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 4980 cgcggtatca ttgcageact ggggccagat ggtaagccct cccgtatcgt agttatetac 5040 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 5100 agcattggta teactgatta 'ac gtcagac CAAG ttact catatatact ttagattgat 5160 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 5220 accaaaatcc cttaacgtga gttttcgttc caetgagcgt cagaccccgt agaaaagatc 5280 aaa gatctt cttgagatcc tttttttctg cgcgtaa.ct gctgcttgca aacaaaaaaa 5340 ccacegctac cagcggtggt ttgtttgccg gatcaagage taccaactct ttttccgaag 5400 gtaactggct teageagage gcasatacca ttctagtsta aatactgtcc gccgtagtta 5460 ggccaccact tcaagaaetc tgtagcaccg cetacat ce tegetetget aatcctgtt to 5520 ccagtg ^ ctg ctgccagtgg cg taagtcg tgtcttaccg ggttggactc aagacgatag 5580 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 5640 cctacacega gagcgaacga actgagatac ctacagcgtg agetatgaga aagcgccacg 5700 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 5760 cgcacgaggg agcttceagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 5B20 cacctctgac ttgagcgtcg atttttgtga tgctsgtcag gggggcggag cctatggaaa 5880 aacgccag to ac cggCctt tttacggttc ctggcctttt gctggccttt tgctcacatg 5940 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgaget 6000 gataccgctc gccgcagccg aacgaccgag cgcagcgagt ggaagcggaa cagtgagcga 6060 g-agcgcccaa tacgcaaacc gcctctcccc gcgcgttggc egatteatta atgcagctgg 6120 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 6180 ctcactcatt aggcacccca ggctttacac tttatgette cggetcgtat g tgtgtgga 6240 attgtga cg gataacaatt tcacacagga aacagctatg accatgatta cgaatttggc 6300 caagtcggcc tctaatacga etcactatag ggagetc 6337 «210 > 59 «211? 1146 - «212 > DNA «213 > Zea mays "400:" 59 aggagcggga atgacegaga gaagcaggag cagctcgccc gagetacegg tggcgccgcg 60 atg agcggt cgccggtgga gaagcctecg ttcactctgg gtcagatcaa gaaggcc c 120 ccgccacact gcttcgagcg ctcggtgctc aagtccttct cgtacgtggt ccacgacctg 180 gtsatcgccg cggcgetcet ctacttcgcg ctggccatca taccggcgct cccaagcccg 240 ctccgctacg ccgcctggcc gctgtactgg atcgcgcagg ggtgcgtgtg caccggcgtg 300 tgggtcatcg cgcacgagtg cggccaccac gccttctcgg actactcgct ectggacgac 360 tggtgctgca gtggtcggcc ctcgtcgcte atggtgccct acttctcgtg gaagtacagc 420 caccggcgcc accactccaa cacggggtcc ctggagcgcg acgaggt tt cgtgcccaag 480 aagaaggagg cgctgccgtg gtacaccccg tacgtgtaca acaacccggt cggccgggtg 540 gtgcacatcg tggtgcagct caccctcggg tggccgctgt acctggcgac caacgcgtcg 600 gggcggccgt accegcgctt egcctgceac ttcgacccct acggccccat- ctacaacgac 660 cgggagcgcg cccagatctt egtctcggac gccggcgtcg tggccgtggc gttcgggctg 720 tacaagctgg cggcggcgtt cggggtctgg tgggtggtgc gcgtgtacge cgtgcogctg 780 ctgatcgtga acgcgtggct ggtgctcatc acctacctgc agcacaccca cccgtcgctc 840 ccccactacg actegagega gtgggactgg ctgcgcggcg cgctggccac catggaccgc 900 gactacg ca tcctcaaccg cgtgttccac aacatcacgg acacgcacgt cgcgcaccac 960 ccatgccgca ctcttctcca ctaccacgcc atggaggcca ccaaggcgat caggcceatc 1020 ctcggcgact actaccactt cgacccgacc cctgtcgcca aggcgacctg gcgcgaggcc 1080 tctacgtcga ggggaatgca gcccgaggae cgcaagggcg tcttctggta caacaagaag 1140 ttctag 1146 It is noted that in relation to this date, the best The method known to the applicant for carrying out said invention is that which is clear from the present description of the invention. Having described the invention as above, the content of the following is claimed as property.

Claims (171)

1. An isolated nucleic acid fragment, characterized in that it comprises a maize oleosin promoter wherein said promoter can be full length or partial and further wherein said promoter comprises a nucleotide sequence substantially corresponding to the nucleotide sequence in any of the SEC ID NOS: 19 or 38-49 or said promoter comprises a fragment or subfragment that is substantially similar and functionally equivalent to any of the nucleotide sequences set forth in SEQ ID NOS: 19 or 38-49.

2. The fragment or subfragment of claim 1, characterized in that said fragment or subfragment hybridizes to the nucleotide sequence set forth in SEQ ID NOS: 19 or 34-49 under moderately stringent conditions.

3. An isolated nucleic acid fragment, characterized in that it encodes a corn stearoyl-ACP delta-9 desaturase substantially corresponding to the nucleotide sequence set forth in any of SEQ ID NOS: 8 and 10 or any functionally equivalent subfragment thereof.

4. A nucleic acid fragment, characterized in that it encodes a corn delta-12 desaturase substantially corresponding to the nucleotide sequence set forth in SEQ ID NO: 2 or any functionally equivalent subfragment thereof.

5. A chimeric gene characterized in that it comprises the nucleic acid fragment of claim 3 or the complement of the nucleic acid fragment of claim 3 operably linked to a suitable regulatory sequence wherein the expression of the chimeric gene results in a corn stearic acid phenotype altered

6. A chimeric gene characterized in that it comprises the nucleic acid fragment of claim 4 or the reverse complement of the nucleic acid fragment of claim 4 operably linked to a suitable regulatory sequence wherein the expression of the chimeric gene results in an altered phenotype of oleic acid of corn.

7. The chimeric gene of claim 5, characterized in that it additionally comprises the nucleic acid fragment of claim 1 or 2.

8. The chimeric gene of claim 6, characterized in that it additionally comprises the nucleic acid fragment of claim 1 or 2.

9. The chimeric gene of claim 5, characterized in that it additionally comprises a shortened intron / exon 1.

10. The chimeric gene of claim 6, characterized in that it additionally comprises a shortened intron / exon 1.

11. The chimeric gene of claim 7, characterized in that it additionally comprises a shortened intron / exon 1.

12. The chimeric gene of claim 8, characterized in that it additionally comprises a shortened intron / exon 1.

13. A chimeric gene, characterized in that it comprises the nucleic acid fragment of claim 3 or the reverse complement thereof and a nucleic acid fragment encoding a corn delta-12 desaturase, any functionally equivalent subfragment thereof or the reverse complement of said fragment or subfragment wherein said subfragment is operably linked and further, wherein the expression of the chimeric gene results in an altered corn oil phenotype.

14. The chimeric gene of claim 13, characterized in that the nucleic acid fragment encoding the corn delta-12 desaturase corresponds substantially to the nucleotide sequence set forth in SEQ ID NO: 2 or any functionally equivalent subfragment thereof.

15. A chimeric gene, characterized in that it comprises the nucleic acid fragment of claim 1 or 2, the nucleic acid fragment of claim 3 or the reverse complement thereof and a nucleic acid sequence encoding a corn delta-12 desaturase , any functionally equivalent subfragment thereof or the inverse complement of said fragment or subfragment wherein said fragments are operably linked and further wherein the expression of the chimeric gene results in an altered corn oil phenotype.

16. The chimeric gene of the claim 15, characterized in that the nucleic acid fragment encoding a corn delta-12 desaturase enzyme corresponds substantially to the nucleotide sequence set forth in SEQ ID NO: 2 or any equivalent subfragment thereof.

17. A chimeric gene, characterized in that it comprises the nucleic acid fragment of claim 3, a nucleic acid sequence encoding a delta-12 desaturase, any functionally equivalent subfragment thereof or the reverse complement of said fragment or subfragment and an intro / exon 1 shortened where the fragment is operably linked and also where the expression of the chimeric gene results in an altered corn oil phenotype.

18. The chimeric gene of claim 17, characterized in that the nucleic acid fragment encoding the delta-12 desaturase corresponds substantially to the nucleotide sequence set forth in SEQ ID NO: 2 or any functionally equivalent subfragment thereof.

19. A chimeric gene, characterized in that it comprises the nucleic acid fragment of claim 1 or 2, the nucleic acid fragment of claim 3 or the reverse complement thereof, a nucleic acid sequence encoding a corn delta-12 desaturase, any functionally equivalent subfragment thereof, or the inverse complement of said fragment and a shortened intron / exon 1 wherein said fragment is operably linked and further wherein the expression of the chimeric gene results in an altered corn oil phenotype.

20. The chimeric gene of the claim 19, characterized in that the nucleic acid fragment encoding the delta-12 desaturase corresponds substantially to the nucleotide sequence set forth in SEQ ID NO: 2 or any functionally equivalent subfragment thereof.

21. A chimeric gene, characterized in that it comprises the nucleic acid fragment of claim 1 or 2, the nucleic acid fragment of claim 3 or the reverse complement thereof, the acid fragment of claim 4 or the complement thereof and a intron / shortened exon 1 wherein said fragments are operably linked and further wherein the expression of the chimeric gene results in an altered corn oil phenotype.

22. A chimeric gene, characterized in that it comprises an isolated nucleic acid fragment encoding a corn delta-12 desaturase substantially corresponding to the nucleotide sequence set forth in SEQ ID NO: 1, a functionally equivalent subfragment thereof or the reverse complement of said fragment or subfragment, or an isolated nucleic acid fragment corresponds substantially to the nucleotide sequence set forth in SEQ ID NO: 58 or 59 or a functionally equivalent subfragment thereof or the reverse complement of said fragment or subfragment, the acid fragment nucleic of claim 1 or 2 and shortened intron / exon 1 wherein said fragment is operably linked and further wherein the expression of the chimeric gene results in an altered corn oleic acid phenotype.

23. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 5.

~ 24. A grain of corn obtained from the plant of claim 23, characterized in that said grain has either a stearic acid content of not less than about 20% of the total oil content or a total saturated content of not less than about 35% of the total oil content.

25. A corn plant or plant part thereof, characterized in that it comprises the chimeric gene of claim 6.

26. A grain of corn obtained from the plant of claim 25, characterized in that said grain has either a stearic acid content of not less than about 60% of the total oil content.

27. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 7.

28. A grain of corn obtained from the plant of claim 27, characterized in that said grain has either a stearic acid content of not less than about 20% of the total oil content or a total saturated content of not less than about 35% of the total oil content.

29. A corn plant or plant part thereof, characterized in that it comprises the chimeric gene of claim 8.

30. A grain of corn obtained from the plant of claim 29, characterized in that said grain has either a stearic acid content of not less than about 60% of the total oil content.

31. A corn plant or plant part thereof, characterized in that it comprises the chimeric gene of claim 9.

32. A grain of corn obtained from the plant of claim 31, characterized in that said grain has either a stearic acid content of not less than about 2D% of the total oil content or a total saturated content of not less than about 35% of the total oil content.

33. A corn plant or plant part thereof, characterized in that it comprises the chimeric gene of claim 10.

34. A grain of corn obtained from the plant of claim 33, characterized in that said grain has either a stearic acid content of not less than about 60% of the total oil content.

35. A corn plant or plant part thereof, characterized in that it comprises the chimeric gene of claim 11.

36. A grain of corn obtained from the plant of claim 35, characterized in that said grain has either a stearic acid content of not less than about 20% of the total oil content or a total saturated content of not less than about 35% of the total oil content.

37. A corn plant or plant part thereof, characterized in that it comprises the chimeric gene of claim 12.

38. A grain of corn obtained from the plant of claim 37, characterized in that said grain has either a stearic acid content of not less than about 60% of the total oil content.

39. A corn plant or plant part thereof, characterized in that it comprises the chimeric gene of claim 13.

40. A grain of corn obtained from the plant of claim 39, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not less than about 30% of the total oil content.

41. A corn plant or plant part thereof, characterized in that it comprises the chimeric gene of claim 14.

42. A grain of corn obtained from the plant of claim 41, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not more than about 30% of the total oil content.

43. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 15.

44. A grain of corn obtained from the plant of claim 43, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not less than about 30% of the total total oil content.

45. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 16.

46. A grain of corn obtained from the plant of claim 45, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not less than about 30% of the total oil content.

47. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 17.

48. A grain of corn obtained from the plant of claim 47, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not less than about 30% of the total oil content.

49. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 18.

50. A grain of corn obtained from the plant of claim 49, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not less than about 30% of the total oil content.

51. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 19.

52. A grain of corn obtained from the plant of claim 51, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not less than about 30% of the total oil content.

53. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 20.

54. A grain of corn obtained from the plant of claim 53, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not less than about 30% of the total oil content.

55. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene of claim 21.

56. A grain of corn obtained from the plant of claim 55, characterized in that said grain has either a stearic acid content of not less than about 30% of the total oil content or a total saturated content of not less than about 30% of the total oil content.

57. A corn plant or plant parts thereof, characterized in that it comprises the chimeric gene selected from claims 5, 7, 9 and 11, and a chimeric gene selected from the chimeric genes of claims 6, 8, 10 and 12

58. A grain of corn obtained from the plant of claim 57, characterized in that said grain has a total saturated content of not less than about 30% of the total oil content and an oleic acid content of not less than about 30% of the content of total oil.

59. A corn plant or parts of the plant thereof, characterized in that it comprises the chimeric gene of claim 22.

60. A grain of corn obtained from the plant of claim 59, characterized in that said grain has an oleic acid content of not less than about 60% of the total oil content.

61. Seeds characterized because they are from the corn plant of claim 23 or 25.

62. Seeds characterized because they are from the maize plant of claim 27.

63. Seeds characterized because they are from the corn plant of claim 29.

64. Seeds characterized because they are from the corn plant of claim 31.

65. Seeds characterized because they are from the corn plant of claim 33.

66 Seeds characterized because they are from the corn plant of claim 35.

67. Seeds characterized because they are from the corn plant of claim 37.

68. Seeds characterized because they are from the corn plant of claim 39.

69. Seeds characterized because they are from the corn plant of claim 41.

70. Seeds characterized because they are from the corn plant of claim 43.

71. Seeds characterized because they are from the corn plant of claim 45.

72. Seeds characterized because they are from the corn plant of claim 47.

73. Seeds characterized because they are from the corn plant of claim 49.

74. Seeds characterized because they are from the corn plant of claim 51.

75. Seeds characterized because they are from the corn plant of claim 53.

76. Seeds characterized because they are from the corn plant of claim 55.

77. Seeds characterized because they are from the corn plant of claim 57.

78. Seeds characterized because they are from the corn plant of claim 59.

79. Oil, characterized in that it is obtained from the grain of the corn plants of claims 23 or 25.

80. Oil characterized in that it is obtained from the grain of the corn plants of claim 27.

81. Oil characterized in that it is obtained from the grain of the corn plants of claim 29.

82. Oil characterized in that it is obtained from the grain of the corn plants of claim 31.

83. Oil characterized in that it is obtained from the grain of the corn plants of claim 33.

84. Oil characterized in that it is obtained from the grain of the corn plants of claim 35.

85. Oil characterized because it is obtained from the grain. of the corn plants of claim 37.

86. Oil characterized in that it is obtained from the grain of the corn plants of claim 39.

87. Oil characterized in that it is obtained from the grain of the corn plants of claim 41.

88. Oil characterized in that it is obtained from the grain of the corn plants of claim 43.

89. Oil characterized in that it is obtained from the grain of the corn plants of claim 45-

90. Oil characterized in that it is obtained from the grain of the corn plants of claim 47.

91. Oil characterized in that it is obtained from the grain of the corn plants of claim 49.

92. Oil characterized in that it is obtained from the grain of the corn plants of claim 51.

93. Oil characterized in that it is obtained from the grain of the corn plants of claim 53.

94. Oil characterized in that it is obtained from the grain of the corn plants of claim 55.

95. Oil characterized in that it is obtained from the grain of the corn plants of claim 57.

96. Oil characterized in that it is obtained from the grain of the corn plants of claim 59.

97. Animal food characterized in that it is derived from the processing of the corn grain of claim 61.

98. Animal feed characterized in that it is derived from the processing of the corn grain of claim 62.

99. Animal food characterized in that it is derived from the processing of the corn grain of claim 63.

100. Animal food characterized in that it is derived from the processing of the corn grain of claim 64.

101. Animal food characterized in that it is derived from the corn grain processing of claim 65.

102. Animal food characterized in that it is derived from the processing of the corn grain of claim 66.

103. Animal food characterized in that it is derived from the processing of the corn grain of claim 67.

104. Animal food characterized in that it is derived from the processing of the corn grain of claim 68.

105. Animal food characterized in that it is derived from the processing of the corn grain of claim 69.

106. Animal food characterized in that it is derived from the processing of the corn grain of claim 70.

107. Animal food characterized in that it is derived from the processing of the corn grain of claim 71.

108. Animal feed characterized in that it is derived from the processing of the corn grain of claim 72.

109. Animal food characterized in that it is derived from the processing of the corn grain of claim 73.

110. Animal food characterized in that it is derived from the processing of the corn grain of claim 74.

111. Animal feed characterized in that it is derived from the processing of the corn grain of claim 75.

112. Animal feed characterized in that it is derived from the processing of the corn grain of claim 76.

113. Animal feed characterized in that it is derived from the processing of the corn grain of claim 77.

114. Animal feed characterized in that it is derived from the processing of the corn grain of claim 78.

115. The use of the oil of claim 79 in food, animal feed, baking or industrial applications.

116. The use of the oil of claim 80 in food, animal feed, baking or industrial applications.

117. The use of the oil of claim 81 in food, animal feed, baking or industrial applications.

118. The use of the oil of claim 82 in food, animal feed, baking or industrial applications.

119. The use of the oil of claim 83 in food, animal feed, baking or industrial applications.

120. The use of the oil of claim 84 in food, animal feed, baking or industrial applications.

121. The use of the oil of claim 85 in food, animal feed, baking or industrial applications.

122. The use of the oil of the claim 86 in food, animal feed, baking or industrial applications.

123. The use of the oil of the claim 87 in food, animal feed, baking or industrial applications.

124. The use of the oil of the claim 88 in food, animal feed, baking or industrial applications.

125. The use of the oil of the claim 89 in food, animal feed, baking or industrial applications.

126. The use of the oil of claim 90 in food, animal feed, baking or industrial applications.

127. The use of the oil of claim 91 in food, animal feed, baking or industrial applications.

128. The use of the oil of claim 92 in food, animal feed, baking or industrial applications.

129. The use of the oil of claim 93 in food, animal feed, baking or industrial applications.

130. The use of the oil of claim 94 in food, animal feed, baking or industrial applications.

131. The use of the oil of claim 95 in food, animal feed, baking or industrial applications.

132. The use of the oil of claim 96 in food, animal feed, baking or industrial applications.

133. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 23 or 25.

134. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 27.

135. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 29.

136. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 31.

137. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 33.

138. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 35.

139. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 37.

140. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 39.

141. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 41.

142. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 43.

143. Products characterized because they are made from hydrogenation, fractionation, interesterification ,. or hydrolysis of the plant grain of claim 45.

144. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 47.

145. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 49.

146. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 51.

147. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 53.

148. Products characterized because they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the grain of the rei indication plant.

149. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 57.

150. Products characterized in that they are made from the hydrogenation, fractionation, interesterification, or hydrolysis of the plant grain of claim 59.

151. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 97.

152. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 98.

153. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 99.

154. A method for improving the quality of the channel of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 100.

155. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 101.

156. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 102.

157. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 103.

158. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 104.

159. A method for improving the quality of the channel of an animal characterized in that it comprises feeding the animal an improved quantity of the carcass quality of the animal feed of claim 105.

160. A method for improving the quality of the channel of an animal characterized in that it comprises feeding the animal an improved quantity of the carcass quality of the animal feed of claim 106.

161. . A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 107.

162. A method for improving the quality of the channel of an animal characterized in that it comprises feeding the animal an improved quantity of the carcass quality of the animal feed of claim 108

163. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved quantity of the carcass quality of the animal feed of claim 109.

164. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 110.

165. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 111.

166. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 112.

167. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved quantity of the carcass quality of the animal feed of claim 113.

168. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 114.

169. A grain of corn, characterized in that it comprises in its genome, the chimeric gene of claim 22, wherein said corn grain has an oil content in the range of about 6% to about 10% based on the dry matter and also , wherein said oil content comprises not less than about 60% oleic acid of the total oil content of the seed.

170. Animal feed characterized in that it is derived from the processing of the corn grain of claim 169.

171. A method for improving the quality of the carcass of an animal characterized in that it comprises feeding the animal an improved amount of the carcass quality of the animal feed of claim 170.