Isolated polynucleotide, polypeptide, recombinant construction, cell, transformed yarrowia sp., methods for transforming a cell, producing a transformed plant, producing yeast, producing polyunsaturated fatty acids and producing at least one polyunsaturated fatty acid, seeds, oils, oilseed plants and food or feed

Abstract

POLYNUCLEOTID, ISOLATED, POLYPEPTIDE, RECOMBINANT CONSTRUCTION, CELL, YARROWIA SP. TRANSFORMED, METHODS TO TRANSFORM A CELL, PRODUCE A TRANSFORMED PLANT, PRODUCE YEAST, PRODUCE POLYINSATURATED FATTY ACIDS AND PRODUCE AT LEAST ONE POLYINSATURATED FATTY ACID, SEEDS, OILS, OIL SEEDS AND FOOD OR FEED PLANTS The present invention deals with isolated nucleic acid fragments and recombinant constructs comprising such fragments encoding a delta-8 desaturase along with a method of making long-chain polyunsaturated fatty acids (PUFAs) this delta-8 desaturase in plants and in oleaginous yeasts.

Description

“ISOLATED POLYNUCLEOTIDE, RECOMBINANT CONSTRUCTION, CELL, YARROWIA SP. TRANSFORMED, METHOD FOR TRANSFORMING A CELL, METHODS FOR PRODUCING A TRANSFORMED PLANT, YEAST AND POLYUNSATURATED FATTY ACIDS, SEEDS, OILS, OIL SEED PLANTS AND FOOD OR FEED” This patent application claims the benefit of Provisional Application US 60 / 583041, filed on June 25, 2004 and Provisional Application US 60 / 624812, filed on November 4, 2004, whose entire contents are incorporated in the present invention by way of of reference. Field of Invention The present invention pertains to a polynucleotide sequence encoding a delta-8 desaturase and the use of this desaturase in the manufacture of long-chain polyunsaturated fatty acids (PUFAs). Background of the Invention Lipids / fatty acids are water-insoluble organic biomolecules that can be extracted from cells and tissues by non-polar solvents such as chloroform, ether or benzene. Lipids have several important biological functions, serving as: (1) structural components of membranes; (2) ways of storing and transporting metabolic fuels; (3) a protective coating on the surface of many organisms; and (4) cell surface components related to cell recognition, species specificity, and tissue immunity. More specifically, polyunsaturated fatty acids (PUFAs) are important components of the cell plasma membrane, where they can be found in such forms as phospholipids and can also be found in triglycerides. PUFAs also serve as precursors to other molecules of importance in humans and animals, including prostacyclins, leukotrienes and prostaglandins. There are two main families of PUFAs (ie, omega-3 fatty acids and omega-6 fatty acids). The human body is capable of producing most of the PUFAs the body needs to function; however, eicosapentaenoic acid (EPA; 20:5, delta-5,8,11,14,17) and docosahexaenoic acid (DHA; 22:6, delta-4,7,10,13,16,19) they cannot be efficiently synthesized by the human body and therefore must be supplied through the diet. Since the human body cannot produce adequate amounts of these PUFAs, these are called essential fatty acids. Because of their important roles in human health and nutrition, EPA and DHA are a topic of much interest as discussed in the present invention. DHA is a fatty acid of the omega-3 series according to the position of the last double bond at the methyl end. It is synthesized through alternate steps of desaturation and elongation. DHA production is important because of its beneficial effect on human health; for example, increased intake of DHA has been shown to be beneficial or have a positive effect in inflammatory disorders (eg rheumatoid arthritis), type II diabetes, hypertension, atherosclerosis, depression, myocardial infarction, thrombosis, some cancers, and in prevention the emergence of degenerative disorders such as Alzheimer's disease. Currently, the main sources of DHA are fish and algal oils. EPA and arachidonic acid (AA or ARA; 20:4, delta-5,8,11,14) are both delta-5 essential fatty acids. EPA belongs to the omega-3 series with five double bonds in the acyl chain, is found in marine foods and is abundant in North Atlantic oily fish. Beneficial or positive effects of increased EPA intake have been shown in patients with coronary heart disease, high blood pressure, inflammation, lung and kidney disease, type II diabetes, obesity, ulcerative colitis, Crohn's disease, anorexia nervosa, burns, osteoarthritis, osteoporosis, attention-deficit / hyperactivity disorder, and early stages of colorectal cancer (see, for example, the review of McColl, J., NutraCos 2(4):35-40(2003)). AA belongs to the omega-6 series with four double bonds. The lack of a double bond at the omega-3 position gives AA properties different from those found in EPA. Eicosanoids produced from AA have strong platelet aggregating and inflammatory properties, whereas EPA-derived eicosanoids have antiplatelet aggregation and anti-inflammatory properties. AA is recognized as the major w-6 fatty acid found in the human brain and is an important component of breast milk and many infant formulas, based on its role in early neurological and visual development. AA can be obtained from certain foods such as meat, fish and eggs, but the concentration is low. Gamma-linolenic acid (GLA; 18:3, delta-6,9,12) is another essential fatty acid found in mammals. GLA is the metabolic intermediate for very long-chain omega-6 fatty acids and for several active molecules. In mammals, the formation of long-chain PUFAs is rate-limited through delta-6 desaturation. Many physiological and pathological conditions such as aging, stress, diabetes, eczema and some infections have been shown to compress the delta-6 desaturation step. Furthermore, GLA is easily catabolized from oxidation and rapid cell division associated with certain disorders, eg cancer or inflammation. As described above, research has shown that several omega fatty acids reduce the risk of coronary heart disease, have a positive effect on child development and in certain mental illnesses, autoimmune diseases and joint complaints. However, although there are many health benefits associated with a diet supplemented with these fatty acids, it is recognized that different PUFAs exert different physiological effects in the body (eg, most notably, the opposing physiological effects of GLA and AA). Thus, production of oils using recombinant media is expected to have several advantages over production from natural sources. For example, recombinant organisms that have preferential traits for oil production can be used, since the host's naturally occurring fatty acid profile can be altered by introducing new biosynthetic pathways into the host and / or by suppressing pathways. unwanted PUFAs, thereby resulting in increased levels of production of desired PUFAs (or conjugated forms thereof) and decreased production of unwanted PUFAs. Optionally, recombinant organisms can provide PUFAs in particular forms that can have specific uses; or oil production can be manipulated such that the ratio of omega-3 to omega-6 fatty acids produced in this way is modified and / or a specific PUFA is produced without significant accumulation of other PUFA products downstream or upstream upstream (downstream or upstream) (eg production of oils comprising ARA and not GLA). The mechanism of PUFA synthesis normally occurs through the delta-6 desaturation pathway. For example, long-chain PUFA synthesis in mammals is predominantly a delta-6 desaturation pathway, where the first step is delta-6 desaturation of LA and ALA to yield GLA and stearidonic acid (STA; 18:4, delta-6,9,12,15), respectively. The elongation and desaturation steps of fatty acids cause the appearance of AA and EPA. Consequently, the genes encoding the components of delta-6 desaturases, delta-6 elongases (also identified as C18 / C20 elongases) and delta-5 desaturases have been cloned from a variety of organisms including higher plants, algae, mosses, fungi, nematodes and humans. Humans can synthesize long-chain PUFAs from the essential fatty acids, linoleic acid (LA; 18:2, delta-9,12) and alpha-linolenic acid (ALA; 18:3, delta-9,12,15 ); LA and ALA must be obtained from the diet. However, the biosynthesis of long-chain PUFAs is somewhat limited and is regulated by dietary and hormonal changes. WO 02 / 26946 (published April 4, 2002) describes isolated nucleic acid molecules encoding FAD4, FAD5, FAD5-2 and FAD6 fatty acid desaturase family members that are expressed in organisms that produce PUFA from long chain, e.g. Thraustochytrium, Pythium irregulare, Schizchytrium and Crypthecodinium. It is indicated that the constructs containing the desaturase genes can be used in any expression system including plants, animals and microorganisms to produce cells capable of producing the long chain PUFAs. WO 98 / 55625 (published December 19, 1998) describes the production of PUFAs by expression of polyketide-like gene synthesis genes in plants. WO 98 / 46764 (published October 22, 1998) describes compositions and methods for preparing long chain fatty acids in plants, parts of plants and in plant cells using the nucleic acid sequences and constructs that encode fatty acid desaturases, including delta-5 desaturases, delta-6 desaturases, and delta-12 desaturases. US 6,075,183 (issued to Knutzon et al. on June 13, 2000) describes methods and compositions for synthesizing long-chain PUFAs. long in plants. US 6,459,018 (issued to Knutzon et al., October 1, 2002) describes a method for producing STA in plant seed using a construct comprising a DNA sequence encoding a delta-6 desaturase. Spychalla et al., (Proc. Natl. Acad. Sci. USA, 94:1142-1147 (1997)) describe the isolation and characterization of a cDNA from C. elegans which, when expressed in Arabidopsis, encodes an acid desaturase fatty acid that can catalyze the introduction of an omega-3 double bond into a fatty acid band of Cis and Czo- An alternative pathway for AA and EPA biosynthesis operates in some organisms (ie, the delta-9 elongase / delta-8 desaturase pathway). At present, LA and ALA are first elongated to eicosadienoic acid (EDA; 20:2, delta-11,14) and eicosatrienoic acid (EtrA; 20:3, delta-11,14,17), respectively, by a delta-9 elongase. Subsequent delta-8 and delta-5 desaturation of these products results in AA and EPA. The delta-8 pathway is present inter alia in euglenoid species where it is the dominant pathway for the formation of PUFAs C 20 . WO 2000 / 34439 (published June 15, 2000) describes amino acid and nucleic acid sequences for delta-5 and delta-8 desaturase enzymes. Based on the information presented in the present invention, it is evident that the delta-8 nucleotide and amino acid sequences of WO 2000 / 34439 are not correct. Wallis et al., (Archives of Biochemistry and Biophysics, 365(2):307-316 (May 15, 1999 )) describe the cloning of a gene that appears to encode a delta-8 desaturase in Euglena gracilis. This appears to be the same sequence described in WO 2000 / 34439. Qi et al., ( Nature Biotechnology, 22(6):739-45 (2004 )) describe the production of long-chain PUFAs using, among other things, an E. gracilis delta-8 desaturase; however, the complete sequence of delta-8 desaturase is not provided. WO 2004 / 057001 (published on July 8, 2004) describes amino acid and nucleic acid sequences for an E. gracilis delta-8 desaturase enzyme. An extensive study of PUFAs from natural sources and chemical synthesis is not sufficient for commercial needs. Therefore, it is of interest to obtain alternative means to allow the production of commercial quantities of PUFAs. Biotechnology offers an attractive route to produce long-chain PUFAs in a safe and cost-effective manner in microorganisms and in plants. With respect to microorganisms, many algae, bacteria, molds and yeasts can synthesize oils in the ordinary course of cellular metabolism. Thus, oil production involves culturing the microorganism in an appropriate culture medium to allow oil synthesis, followed by separation of the microorganism from the fermentation medium and treatment to recover the intracellular oil. Attempts have been made to optimize the production of fatty acids by fermentation media which involve varying such parameters as the microorganisms used, the media and the conditions which allow oil production. However, these efforts have proved mostly unsuccessful in improving oil yield or the ability to control the compositional characteristics of produced oil. One type of microorganism that has not been previously examined as a production platform for PUFAs (prior to the work of the filed patent application holders), however, is oleaginous yeast. These organisms can accumulate oil up to 80% of their cellular dry weight. The technology for the growth of oleaginous yeasts with high oil content is well developed (eg, see EP 0,005,277 B1; Ratledge, C., Prog. Ind. Microbiol. 16:119-206 (1982)) and may offer a cost advantage compared to microalgae fermentation commercial products for the production of omega-3 or omega-6 PUFAs. Whole yeast cells may also represent a convenient way to encapsulate PUFAs rich in omega-3 or omega-6 oils for use in functional foods and animal feed supplements. Documents WO 2004 / 101757 and WO 2004 / 101753 (published on November 25, 2004) refer to the production of PUFAs in oleaginous yeasts and are copending applications of the holders of the filed patent application. WO 2004 / 071467 (published on August 6, 2004) refers to the production of PUFAs in plants, whereas WO 2004 / 071178 (published on August 26, 2004) refers to annexin and their use in transgene expression in plants; both of which are co-pending applications of the holders of the filed patent application. Brief Description of the Invention The present invention relates to an isolated polynucleotide comprising: (a) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, the polypeptide having an amino acid sequence consisting essentially of SEQ ID NO:2 or 113; or, (b) a complement of the nucleotide sequence, where the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary. In a second embodiment, the present invention relates to the construction of a recombinant comprising SEQ ID NO: 1 or 112 operably linked to at least one regulatory sequence. In a third embodiment, the present invention relates to a cell comprising the recombinant construct of the present invention. In a fourth embodiment, the present invention relates to a method for transforming cells, plants and yeast with the recombinant construct of the present invention. In a fifth embodiment, the present invention relates to seeds obtained from such plants and oil obtained from such seeds. In a sixth embodiment, the present invention relates to a method of manufacturing polyunsaturated fatty acids in a cell. In a seventh embodiment, the present invention relates to an oilseed plant comprising a first recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide operably linked to at least one sequence regulator; and at least one additional recombinant DNA construct comprising an isolated polynucleotide operably linked to at least one regulatory sequence encoding a polypeptide selected from the group consisting of delta-4, delta-5, delta-6, delta-9, delta-12, delta-15 and delta-17 desaturase, a delta-9 elongase, a Ci elongase 8 to C22 and a C20 to C24 elongase. In yet another aspect, the present invention relates to a method for producing at least one polyunsaturated fatty acid in a soybean cell, which comprises: (a) transforming a soybean cell with a first recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide operably linked to at least one regulatory sequence and at least one additional recombinant DNA construct comprising an isolated polynucleotide operably linked to at least one regulatory sequence encoding a polypeptide selected from the group consisting of delta-4, delta-5 , delta-6, delta-9, delta-12, delta-15 and delta-17 desaturase, a delta-9 elongase, a C elongase 18 to C22 and an elongase C20 to C24- (b) regenerating a soybean plant from the transformed cell of step (a); and (c) selecting seeds obtained from plants in step (b) that have an altered level of polyunsaturated fatty acids when compared to the level in seeds obtained from a non-transformed soybean plant. In an eighth embodiment, the present invention relates to an oilseed plant selected from the group consisting of soybean, Brassica species, sunflower, corn, cotton, linseed and saffron. In a ninth embodiment, the present invention relates to oilseed plants in which the polyunsaturated fatty acid is selected from the group consisting of AA, EDA, EPA, ETA, EtrA, DGLA, DPA and DHA. Additional embodiments include seeds and oil obtained from plants transformed with the isolated polynucleotides of the present invention. Additional embodiments relate to food, feed and ingredients derived from the processing of seeds obtained from plants transformed with the isolated polynucleotides of the present invention. Biological Deposits The following plasmids have been deposited with the American Type Culture Collection (ATCC), 10801 University Boulevard, Manassas, VA 20110-2209 and bear the following designations, accession numbers, and deposit dates. Plasmid Accession Number Date of Deposit pKR681 ATCC PTA-6046 June 4, 2004 pKR685 ATCC PTA-6047 June 4, 2004 pY89-5 ATCC PTA-6048 June 4, 2004 pKR274 ATCC PTA-4988 January 30, 2003 PKR669 June 13, 2005 PKR786 June 13, 2005 Brief Description of Figures and Sequence Lists The present invention can be more fully understood from the following detailed description and the accompanying drawings and sequence listing, which form a part of the present application. Sequence descriptions summarize the listings of sequences appended to the sequence. The sequence listing contains one letter codes for nucleotide sequence characters and one and three letter codes for amino acids as defined in the IUPAC-IUB standards described in Nucleic Acids Research 13:3021-3030 (1985) and in Biochemical Journal 219(2):345-373(1984). Figure 1 shows a chromatogram of the lipid profile of a cell extract of Euglena gracilis as described in Example 10. Figure 2 shows a polypeptide sequence alignment of the claimed delta-8 desaturase from Euglena gracilis (SEQ ID NO:2), a version of a delta-8 desaturase with reduced activity (SEQ ID NO:4) and non published functionalities of delta-8 desaturase sequences determined in gi:5639724 (GenBank Accession No. AAD45877 and SEQ ID NO:6) and WO 00 / 34439 or Wallis et al., (Archives of Biochem. Biophys, 365:307-316 (1999)) (SEQ ID NO:7). The alignment method used corresponds to the "Clustal V alignment method". Figure 3 provides plasmid maps for the following: (A) yeast expression vector pY89-5 as described in Example 5; and (B) soy expression vector pKR681 as described in Example 6. Figure 4 provides plasmid maps for the following: (A) pKR685 soy expression vector as described in Example 8; and (B) pKR274 expression vector as described in Example 9. Figure 5 provides plasmid maps for the following: (A) pDMW240 yeast expression vector as described in Example 1; (B) pDMW255 yeast expression vector as described in Example 1; (C) pDMW261 yeast expression vector as described in Example 1; and, (D) pKUNFmKF2 vector as described in Example 14. Figure 6 provides plasmid maps for the following: (A) yeast expression vector pDMW277 as described in Example 14; (B) pZF5T-PPC vector as described in Example 14; (C) pDMW287 yeast expression vector as described in Example 14; and (D) yeast expression vector pDMW287F as described in Example 14. Figure 7 provides plasmid maps for the following: (A) pZUF17 vector as described in Example 15; (B) pDMW237 yeast expression vector as described in Example 15; (C) pKUNT2 yeast expression vector as described in Example 16; and, (D) pDMW297 yeast expression vector as described in Example 16. Figure 8 provides plasmid maps for the following: (A) pKR682 soy expression vector as described in Example 17; (B) pKR786 soy expression vector as described in Example 18; and (C) pKR669 soy expression vector as described in Example 19. Figure 9 is a representative biosynthetic pathway of PUFAs. Figure 10 shows a chromatogram of the lipid profile of a soybean embryo extract as described in Example 22. SEQ ID NO:1 represents 1271 bp of Euglena gracilis sequence which contains the ORF (nucleotides 4-1269 (of Stop)) of the delta-8 desaturase gene. SEQ ID NO:2 is the amino acid sequence encoded by nucleotides 4-1269 of SEQ ID NO:1. SEQ ID NO:3 represents 1271 bp of sequence from Euglena gracilis which contains the ORF (nucleotides 4-1269 (Stop)) of the delta-8 desaturase gene which contains a guanine to adenine substitution at position 835, in comparison to the sequence of SEQ ID NO:1. SEQ ID NO:4 is the deduced amino acid sequence encoded by nucleotides 4-1269 of SEQ ID NO:3, which contains an alanine for threonine substitution at position 278 as compared to the polypeptide sequence of SEQ ID NO: :two. SEQ ID NO:5 represents 1275 bp of the Euglena gracilis sequence determined at gi:5639724 (GenBank Accession No. AAD45877), which contains the ORF (nucleotides 14-1273 (for Stop)) of a non-functional version of delta-8 desaturase gene. SEQ ID NO:6 is the nucleotide encoded deduced amino acid sequence of SEQ ID NO:5 and given in gi:5639724. SEQ ID NO:7 is the amino acid sequence of a non-functional version of delta-8 desaturase described in Wallis et al., (Archives of Biochem. Biophys., 365:307-316 (1999) and WO 00 / 34439). SEQ ID NO:8 is the forward primer used for the amplification of Euglena gracilis delta-8 desaturase in Example 3. SEQ ID NO:9 is the reverse primer used for the amplification of Euglena gracilis delta-8 desaturase in Example 3. SEQ ID NO:10 is the forward primer used to sequence a delta-8 desaturase clone as described in Example 3. SEQ ID NO:11 is the reverse primer used to sequence a delta-8 desaturase clone as described in Example 3. SEQ ID NO:12 is the forward primer used to sequence a delta-8 desaturase clone as described in Example 3. SEQ ID NO:13 is the reverse primer used to sequence a delta-8 desaturase clone as described in Example 3. SEQ ID NO:14 is the multiple restriction enzyme site sequence inserted in front of the beta-conglycinin promoter as described in Example 6. SEQ ID NO:15 is the forward primer used for elongase amplification. SEQ ID NO:16 is the reverse primer used for elongase amplification. SEQ ID NO:17 is the multiple restriction enzyme site sequence inserted upstream of the Kti promoter as described in Example 6. SEQ ID NO:18 determines the transcription termination sequence of soybean albumin with restriction enzyme sites as described in Example 6. SEQ ID NO:19 is the oSalb-12 primer used for transcription end amplification of albumin. SEQ ID NO:20 is the oSalb-13 primer used for transcription end amplification of albumin. SEQ ID NO:21 is the GSP1 primer used for amplification of the soybean annexin gene. SEQ ID NO:22 is the GSP2 primer used for amplification of the soybean annexin gene. SEQ ID N°:23 is the GSP3 primer used for the amplification of BD30 soy. SEQ ID N°:24 is the GSP4 primer used for the amplification of the BD30 soy. SEQ ID NO:25 determines the BD30 soy promoter sequence. SEQ ID NO:26 determines the soybean glycinin Gy1 promoter sequence. SEQ ID NO:27 is the forward primer used for amplification of the soybean glycinin Gy1 promoter sequence. SEQ ID NO:28 is the reverse primer used for amplification of the soybean glycinin Gy1 promoter sequence. SEQ ID NO:29 determines the sequence of the soybean annexin promoter. SEQ ID NO:30 is the forward primer used for amplification of the soybean annexin promoter sequence. SEQ ID NO:31 is the reverse primer used for amplification of the soybean annexin promoter sequence. SEQ ID NO:32 is the forward primer used for amplification of the BD30 soy promoter sequence. SEQ ID NO:33 is the reverse primer used for amplification of the BD30 soybean promoter sequence. SEQ ID NO:34 is the oKTi6 primer used for the amplification of the Kti / A / ofZ / Kti 3' cassette. SEQ ID NO:35 is the 0KTI6 primer used for the amplification of the Kti / A / ofZ / Kti 3' cassette. SEQ ID NO:36 is the oSBD30-1 primer used for amplification of the 3' soybean BD30 transcription terminus. SEQ ID NO:37 is the oSBD30-2 primer used for amplification of the 3' soybean BD30 transcription terminus. SEQ ID NO:38 is the oCGR5-1 primer used for the amplification of M. alpina delta-6 desaturase. SEQ ID NO:39 is the oCGR5-2 primer used for the amplification of M. alpina delta-6 desaturase. SEQ ID NO:40 is the oSGIy-1 primer used for amplification of the glycinin Gy1 promoter. SEQ ID NO:41 is the oSGIy-2 primer used for amplification of the glycinin Gy1 promoter. SEQ ID NO:42 is the LegPro5' primer used for amplification of the legA2 promoter sequence. SEQ ID NO:43 is the LegPro3' primer used for the amplification of the legA2 promoter sequence. SEQ ID NO:44 is the LegTermõ' primer used for leg2A transcription termination amplification. SEQ ID NO:45 is the LegTerm3' primer used for leg2A transcription termination amplification. SEQ ID NO:46 is the CGR4 forward primer used for the amplification of M. alpina desaturase. SEQ ID NO:47 is the CGR4 reverse primer used for the amplification of M. alpina desaturase. SEQ ID N°:48 is the sequence from Euglena gracilis, determined at nucleotides 14-1275 of SEQ ID N°:5, optimized for codon usage in Yarrowia lipolytica. SEQ ID NO:49-74 correspond to primers D8-1A, D8-1B, D8-1B, D8-2A, D8-2B, D8-3A, D8-3B, D8-4A, D8-4B, D8- 5A, D8-5B, D8-6A, D8-6B, D8-7A, D8-7B, D8-8A, D8-8B, D8-9A, D8-9B, D8-10A, D8-10B, D8-11A, D8-11B, D8-12A, D8-12B, D8-13A, and D8-13B, respectively, used for amplification as described in Example 1. SEQ ID NO. os :75-82 correspond to primers D8-1F, D8-3R, D8-4F, D8-6R, D8-7F, D8-9R, D8-10F and D8-13R, respectively, used for amplification as described in Example 1. SEQ ID NO:83 is the 309 bp Nco / Bglll fragment described in Example 1. SEQ ID NO:84 is the 321 bp BgIII / XhoI fragment described in Example 1. SEQ ID NO:85 is the 264 bp Xhol / SacI fragment described in Example 1. SEQ ID NO:86 is the 369 bp Sac1 / Not1 fragment described in Example 1. SEQ ID NO:87 is primer ODMW390 used for amplification as described in Example 1. SEQ ID NO:88 is primer ODMW391 used for amplification as described in Example 1. SEQ ID NO:89 is the chimeric gene described in Example 1. SEQ ID NO:90 is the chimeric gene described in Example 1. SEQ ID NO:91 is primer ODMW392 used for amplification as described in Example 1. SEQ ID NO:92 is primer ODMW393 used for amplification as described in Example 1. SEQ ID NO:93 is the synthetic delta-8 desaturase described in Example 1. SEQ ID NO:94 is primer ODMW404 used for amplification as described in Example 14. SEQ ID NO:95 is the Kpn / Not1 fragment described in Example 14. SEQ ID NO:96-111 correspond to primers YL521, YL522, YL525, YL526, YL527, YL528, YL529, YL530, YL531, YL532, YL533, YL534, YL535, YL536, YL537 and YL538, respectively, used for amplification as described in Example 14. SEQ ID NO:112 is the nucleotide sequence for the synthetic delta-8 desaturase codon-optimized for expression in Yarrowia lipolytica. SEQ ID NO:113 is the amino acid sequence encoded by nucleotides 2-1270 of SEQ ID NO:112. SEQ ID NO:114 is the DNA sequence (995 bp) of the Yarrowia lipolytica fructose bisphosphate aldolase promoter containing a Yarrowia Intron (FBAIN). SEQ ID NO:118 is the nucleotide sequence for synthetic delta-9 elongase codon-optimized for expression in Yarrowia lipolytica. SEQ ID NO:119 is the DNA sequence of delta-9 elongase from Isochrysis galbana (792 bp), SEQ ID NO:120 is the amino acid sequence of delta-9 elongase from Isochrysis galbana (263 AA ). SEQ ID NO:121-136 correspond to primers IL3-1A, IL3-1B, IL3-2A, IL3-2B, IL3-3A, IL3-3B, IL3-4A, IL3-4B, IL3-5A, IL3- 5B, IL3-6A, IL3-6B, IL3-7A, IL3-7B, IL3-8A and IL3-8B, respectively, used for amplification as described in Example 15. SEQ ID NO: 137-140 correspond to primers IL3-1F, IL3-4R, IL3-5F and JL3-8R, respectively, used for amplification as described in Example 15. SEQ ID NO:141 is the 417 bp NcoI / PstI fragment described in Example 15. SEQ ID NO:142 is the 377 bp PstI / Not1 fragment described in Example 15. SEQ ID N°:146 is the DNA sequence of delta-12 desaturase from Yarrowia lipolytica (1936 bp), SEQ ID N°:147 is the amino acid sequence of delta-12 desaturase from Yarrowia lipolytica (419 AA ). SEQ ID NO:149 is the olGsel1-1 primer used for the amplification of a delta-9 elongase as described in Example 17. SEQ ID NO:150 is the olGsel1-2 primer used for the amplification of a delta-9 elongase as described in Example 17. SEQ ID NO:151 is the fragment described in Example 18. SEQ ID NO: 115, 116, 117, 143, 144, 145 and 148 are plasmids as identified in Table 1. Table 1 Summary of Plasmid SEQ ID Numbers Plasmid SEQ ID No. Length pY54PC 115 8502 bp pKUNFmkF2 116 7145 bp pZF5T-PPC 117 5553 bp pZUF17 143 8165 bp pDMW237 144 7879 pKUNT2 145 6457 bp pDMW290 1448 bp Detailed Description of the Invention All patents, publications and patent applications cited in the present invention are incorporated by reference in their entirety. In the context of this description, a number of terms will be used. Definitions The term "fatty acids" refers to long-chain aliphatic acids (alkanoic acids) of varying chain lengths from about C12 to C22 (although acids with a shorter chain length are known). The predominant chain lengths are between C16 and C 22 . Additional details regarding the differentiation between "acids "saturated fatty acids" versus "unsaturated fatty acids", "monounsaturated fatty acids" versus "polyunsaturated fatty acids" (or "PUFAs") and "omega-6 fatty acids" (co-6 or n-6) versus "omega fatty acids" -3" (w-3 or n-3) are provided in WO 2004 / 101757. Fatty acids are described in the present invention by a simple "X:Y" notation system, where the number before the colon indicates the number of carbon atoms in the fatty acid and the number after the colon is the number of double bonds that are present. The number following the fatty acid designation indicates the position of the carboxyl-terminal double bond of the fatty acid with the "c" affix to the cis configuration of the double bond [e.g., palmitic acid (16:0), stearic acid ( 18:0), oleic acid (18:1, 9c), petroselinic acid (18:1, 6c), LA (18:2, 9c, 12c), GLA (18:3, 6c, 9c, 12c) and ALA (18:3, 9c, 12c, 15c)]. Unless otherwise specified, 18:1, 18:2 and 18:3 refer to oleic, LA and linolenic fatty acids. If not specifically written otherwise, the double bonds are assumed to be of cis configuration. For example, the double bonds in 18:2 (9.12) are assumed to be in the cis configuration. A representative pathway is illustrated in Figure 9, which provides the conversion of stearic acid through various intermediates to DHA, which demonstrates how w-3 and o>-6 fatty acids can be produced from a common source. The nomenclature used to describe PUFAs in the present description is shown below in Table 2. In the column entitled "Shorthand Notation", the omega reference system is used to indicate the number of carbons, the number of double bonds and the position of the bond double closest to the omega carbon, counting from the omega carbon (which is numbered 1 for this purpose). The remainder of the table summarizes the names names of omega-3 and omega-6 fatty acids, whose abbreviations will be used throughout the remainder of the specification and each chemical name of the compound. Table 2 Nomenclature of Polyunsaturated Fatty Acids Common Name Abbreviation Chemical Name Shorthand Notation Linoleic LA c / s-9,12-octadecadienoic 18:2 w-6 y-Linoleic GLA c / s-6, 9,12-octadecatrienoic 18:3 w-6 Eicosadienoic EDA c / s-11,14-eicosadienoic 20:2 w-6 Dihomo-y-linoleic DGLA c / s-8,11,14-eicosatrienoic 20:3 w-6 Arachidonic Aa or ARA c / s-5, 8,11, 14-eicosatetraenoic 20:4 w-6 a-Linolenic ALA c / s-9,12,15-octadecatrienoic 18:3 w-3 Stearidonic STA c / s-6, 9,12,15-octadecatetraenoic 18:4 w- 3 Eicosatrienoic ETrA c / s-11,14,17-eicosatrienoic 20:3 w-3 Eicosatetraenoic ETA c / s-8,11,14,17-eicosatetraenoic 20:4 w-3 Eicosapentaenoic EPA c / s-5, 8 ,11,14,17-eicosapentaenoic 20:5 w-3 Docosapentaenoic DPA w / s-7,10,13,16,19-docosapentaenoic 22:5 w-3 Docosahexaenoic DHA w / s-4, 7,10,13 ,16,19-docosahexaenoic 22:6 w-3 The term "essential fatty acid" refers to a specific PUFA that an organism must ingest in order to survive, being unable to synthesize the specific essential fatty acid again. For example, mammals cannot synthesize the essential fatty acid LA. Other essential fatty acids include GLA, DGLA, ARA, EPA and DHA. The term "fat" refers to a lipid substance that is solid at 25°C and generally saturated. The term "oil" refers to a lipid substance that is liquid at 25°C and generally polyunsaturated. PUFAs are found in the oils of some algae, oleaginous yeasts and filamentous fungi. "Microbial oils" or "single-cell oils" are those oils produced naturally by microorganisms during their lifetime. Such oils may contain the long chain PUFAs. The term "PUFA biosynthetic pathway" refers to a metabolic process that converts oleic acid to LA, EDA, GLA, DGLA, ARA, ALA, STA, ETrA, ETA, EPA, DPA, and DHA. This process is well described in the literature (for example see document WO 2005 / 003322). Simply put, this process involves elongating the carbon chain by adding carbon atoms and desaturating the molecule by adding double bonds, using a series of special desaturation and elongation enzymes (i.e., "enzymes"). of the PUFA biosynthetic pathway") present in the membrane of the endoplasmic reticulum. More specifically, "PUFA biosynthetic pathway enzymes" refer to any of the following enzymes (and the genes encoding said enzymes) associated with the biosynthesis of a PUFA, including: delta-4 desaturase, delta-5 desaturase , delta-6 desaturase, delta-12 desaturase, delta-15 desaturase, delta-17 desaturase, delta-9 desaturase, delta-8 desaturase, elongase C14 / C 16 , elongase C16 / C18, elongase C 18 / Ç 2 o and / or elongase 620 / 622- A "desaturase" is a polypeptide that can desaturate one or more fatty acids to produce a mono- or polyunsaturated fatty acid or precursor of interest. Of particular interest in the present invention are delta-8 desaturases which desaturate a fatty acid between the eighth and ninth numbered carbon atom of the carboxyl terminal end of the molecule and which can, for example, catalyze the conversion of EDA to DGLA and / or ETrA in ETA. Other useful fatty acid desaturases include, for example: (1) delta-5 desaturases which catalyze the conversion of DGLA to ARA and / or ETA in EPA; (2) delta-6 desaturases that catalyze the conversion of LA to GLA and / or ALA to STA; (3) delta-4 desaturases that catalyze the conversion of DPA to DHA; (4) delta-12 desaturases that catalyze the conversion of oleic acid to LA; (5) delta-15 desaturases that catalyze the conversion of LA to ALA and / or GLA to STA; (6) delta-17 desaturases that catalyze the conversion of ARA to EPA and / or DGLA to ETA; and (7) delta-9 desaturases which catalyze the conversion of palmitate to palmitoleic acid (16:1) and / or stearate to oleic acid (18:1). The term "elongase system" refers to a set of four enzymes that are responsible for elongating a fatty acid carbon chain to produce a fatty acid that is two carbons longer than the fatty acid substrate on which the elongase system. More specifically, the elongation process occurs in association with fatty acid synthase, whereby CoA is the acyl carrier (Lassner et al., Plant Cell 8:281-292 (1996)). In the first step, which was found to be substrate-specific and also rate-limiting, malonyl-CoA is condensed with a long-chain acyl-CoA to yield CO2 and p-ketoacyl-CoA (where the acyl moiety has been elongated by two carbon atoms). Subsequent reactions include reduction to p-hydroxyacyl-CoA, dehydration to an enoyl-CoA, and a second reduction to yield the elongated acyl-CoA. Examples of reactions catalyzed by elongase systems include the conversion of GLA to DGLA, STA to ETA, and EPA to DPA. For purposes of the present invention, an enzyme that catalyzes the first condensation reaction (i.e., conversion of malonyl-CoA to p-ketoacyl-CoA) will be referred to generically as an "elongase". In general, the substrate selectivity of elongases is somewhat broad but segregated by chain length and degree of unsaturation. Consequently, elongases can have different specificities. Per For example, a C16 / C18 elongase will use a substrate (e.g. palmitate), an 0^620 elongase will use a Cie substrate (e.g. GLA, STA) and a C20 / C22 elongase will use a C20 substrate (e.g. , EPA). Similarly, a delta-9 elongase can catalyze the conversion of LA and ALA to EDA and ETrA, respectively (see WO 2002 / 077213). It is important to note that some elongases have a broad specificity and thus a single enzyme may be able to catalyze multiple elongase reactions (eg, thereby acting as both a C-ie / C-is elongase and a Cys / C2o elongase)- The term "delta-9 elongase / delta-8 desaturase pathway" refers to a biosynthetic pathway for the production of long-chain PUFAs, said pathway comprising at least one delta-9 elongase and one delta-8 desaturase and thereby permits the biosynthesis of DGLA and / or ETA from LA and ALA, respectively. This route may be advantageous in some embodiments, since STA and / or STA biosynthesis is excluded. The terms "polynucleotide", "polynucleotide sequence", "nucleic acid sequence", "nucleic acid fragment" and "isolated nucleic acid fragment" are used interchangeably in the present invention. These terms encompass nucleotide sequences and the like. A polynucleotide may be a single or double stranded RNA or DNA polymer, which optionally contains synthetic unnatural or altered nucleotide bases. A polynucleotide in the form of a DNA polymer can be comprised of one or more segments of cDNA, genomic DNA, synthetic DNA or mixtures thereof. Nucleotides (commonly found in their 5'-monophosphate form) are named by a single letter designation as follows: "A" for adenylate or deoxyadenylate (for RNA or DNA, respectively), "C" for the cytidylate or deoxycytidylate, "G" for guanylate or deoxyguanylate, "U" for uridylate, "T" for deoxythymidylate, "R" for purines (A or G), "Y" for pyrimidines (C or T), " K" for G or T, "H" for A or C or T, "I" for inosine, and "N" for any nucleotide. The terms "subfragment that is functionally equivalent" and "functionally equivalent subfragment" are used interchangeably in the present invention. These terms refer to a portion or a subsequence of an isolated fragment of nucleic acid in which the ability to alter gene expression or to produce a particular phenotype is retained whether the fragment or subfragment encodes an active enzyme or not. For example, the fragment or subfragment can be used in the design of chimeric genes to produce the desired phenotype in a transformed plant. Chimeric genes can be designed for use in deletion by linking a nucleic acid fragment or a subfragment thereof, whether it encodes an active enzyme or not, in sense or antisense orientation relative to a plant promoter sequence. The terms "homology", "homologous", "substantially similar" and "substantially corresponding" are used interchangeably in the present invention. They refer to nucleic acid fragments in which changes in one or more bases of the nucleotide do not affect the ability of the nucleic acid fragment to mediate gene expression or to produce a particular phenotype. These 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 should be understood, therefore, as those skilled in the art will appreciate, that the present invention encompasses more than the specific example sequences. Furthermore, those skilled in the art recognize that substantially similar nucleic acid sequences encompassed by the present invention are also defined by their ability to hybridize (under moderately stringent conditions, e.g., 0.5X SSC, 0.1% SDS, 60 °C) with the sequences exemplified in the present invention or for any portion of the nucleotide sequences described in the present invention and which are functionally equivalent to any of the nucleic acid sequences described in the present invention. Stringent conditions can be adjusted to select from moderately similar fragments, such as homologous sequences from not closely related organisms, to highly similar fragments, such as genes that duplicate functional enzymes from closely related organisms. Post-hybridization washes determine stringent conditions. One set of preferred conditions involves a series of washes starting with 6X SSC, 0.5% SDS at room temperature for 15 minutes, then repeated with 2X SSC, 0.5% SDS at 45°C for 30 minutes and repeated then twice with 0.2X SSC, 0.5% SDS at 50°C for 30 minutes. A preferred set of stringent conditions involves using higher temperatures where the washes are identical to those above except that the temperature of the final two 30 minute washes in 0.2X SSC, 0.5% SDS was increased to 60° Ç. Another preferred set of highly stringent conditions involves the use of two final washes in 0.1X SSC, 0.1% SDS at 65°C. "Gene" refers to a fragment of nucleic acid that expresses a specific protein, including the regulatory sequences that precede (5' non-coding sequences) and follow (3' non-coding sequences) the coding sequence. The "native gene" refers to a gene as found in nature with its own regulatory sequences. The "chimeric gene" refers to any gene that is not a native gene, which comprises coding and regulatory sequences that are not found together in nature. Consequently, a chimeric gene may comprise regulatory sequences and coding sequences that are derived from different sources or regulatory sequences and coding sequences that are derived from the same source but that are arranged in a different manner than that found in the nature. An "exogenous" gene refers to a gene not normally found in the host organism, but which is inserted into the host organism by gene transfer. Exogenous genes can comprise native genes inserted into a non-native organism or chimeric genes. A "transgene" is a gene that is inserted into the genome by a transformation procedure. A "codon-optimized gene" is a gene that has its codon usage frequency designed to mimic the host cell's preferred codon usage frequency. An "allele" is one of several alternative forms of a gene that occupies a given locus on a chromosome. When all alleles present at a given locus on a chromosome are the same, that plant is homozygous at that locus. If the alleles present at a particular locus on a chromosome differ, that plant is heterozygous at that locus. The "coding sequence" refers to a DNA sequence that encodes a specific sequence of amino acids. "Regulatory sequences" refer to nucleotide sequences found upstream (5' non-coding sequences), within, or downstream (3' non-coding sequences) of a coding sequence and which influence transcription, processing either the stability of the RNA or the translation of the associated coding sequence. Regulatory sequences can include, but are not limited to: promoters, translation leader sequences, introns, polyadenylation recognition sequences, RNA processing sites, effector binding sites and stem-loop structures. The "promoter" refers to a DNA sequence capable of controlling the expression of a coding sequence or a functional RNA. The promoter sequence consists of proximal and more distal upstream elements, with the latter elements often referred to as enhancers. Accordingly, an "enhancer" is a DNA sequence that can stimulate a promoter's activity and can be either an innate promoter element or a heterologous element inserted to enhance the level or tissue specificity of a promoter. Promoters may be derived entirely from a native gene or may be composed of different elements derived from different promoters found in nature or even comprise synthetic segments of DNA. It should be understood by those skilled in the art that different promoters can 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. It should also be recognized that, since in most cases the exact boundaries of the regulatory sequences have not been fully defined, DNA fragments of some variations may have identical promoter activity. The promoters that cause a gene to be expressed in most cell types most of the time are generally referred to as "constitutive promoters". New promoters of various types useful in plant cells are constantly being discovered; numerous examples can be found in the compilation made by Okamuro, J.K. and Goldberg, R.B. Biochemistry of Plants 15:1-82 (1989). The "translation leader sequence" refers to a sequence of polynucleotides located between the promoter sequence of a gene and the coding sequence. The translation leader sequence is present in the fully processed mRNA upstream of the translation initiation sequence. The translation leader sequence can affect processing from primary transcript to mRNA, mRNA stability, or translation efficiency. Examples of translation leader sequences have been described (Turner, R. and Foster, G. D'Mol. Biotechnol. 3:225-236 (1995)). "3' non-coding sequences", "transcriptional termination" or "termination sequences" refer to DNA sequences located downstream of a coding sequence and include polyadenylation recognition sequences and other sequences that encode regulatory signals capable of affecting mRNA processing or gene expression. The polyadenylation signal is generally characterized by affecting the addition of polyadenylic acid systems to the 3' end of the mRNA precursor. The use of different 3' non-coding sequences is exemplified by Ingelbrecht, I.L, et al., Plant Cell 1:671-680(1989). "RNA transcript" refers to the product resulting from RNA polymerase-catalyzed transcription of a DNA sequence. When the RNA transcript is a perfect complementary copy of the DNA sequence, it is termed as a primary transcript. An RNA transcript is termed as mature RNA when it is an RNA sequence derived from post-transcriptional processing of the primary transcript. "Messenger RNA" or "mRNA" refers to RNA that does not have introns and that can be translated into protein by the cell. "cDNA" refers to DNA that is complementary to and synthesized from a template strand of the mRNA by using enzymatic reverse transcriptase. The cDNA can be single-stranded or can be converted into double-stranded form by using the Klenow fragment of DNA polymerase I. "Sense" RNA refers to the RNA transcript that includes the mRNA and can be translated into protein within a cell or in vitro. "Antisense RNA" refers to an RNA transcript that is complementary to all or part of a primary transcript or target mRNA and that blocks expression of a target gene (US 5,107,065). The complementarity of an antisense RNA can be with any part of the gene-specific transcript, ie the 5' non-coding sequence, the 3' non-coding sequence, the introns or the coding sequence. "Functional RNA" refers to antisense RNA, ribozyme RNA, or other RNA that may not be translated but still has an effect on cellular processes. The terms "complement" and "reverse complement" are used interchangeably in the present invention with respect to transcribed mRNA and lend themselves to defining the antisense RNA of the message. The term "operably linked" refers to the association of nucleic acid sequences into a single nucleic acid fragment such that the function of one is regulated by the other. For example, a promoter is operably linked to a coding sequence when it is capable of regulating the expression of that coding sequence (ie, the coding sequence is under the transcriptional control of the promoter). The coding sequences can be operably linked to the regulatory sequences in a sense or antisense orientation. In another example, the complementary regions of the RNA of the present invention can be operably linked, directly or indirectly, 5' to the target mRNA or 3' to the target mRNA or within the target mRNA or a first complementary region is 5' and its complement is 3' to the target mRNA. Standard recombinant DNA and molecular cloning techniques used in the present invention are well known in the art and described more fully in Sambrook, J., Fritsch, E.F. and Maniatis, T. Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory: Cold Spring Harbour, NY (1989). Transformation methods are well known to those skilled in the art and described below. "PCR" or the "Polymerase Chain Reaction" is a technique for the synthesis of large amounts of specific segments of DNA and consists of a series of repetitive cycles (Perkin Elmer Cetus Instruments, Norwalk, CT). Typically, double-stranded DNA is heat-denatured, with the two primers complementary to the 3' boundaries of the target segment annealed at low temperature and then extended at an intermediate temperature. A set of these three consecutive steps is termed as a "cycle". The term "recombinant" refers to an artificial combination of two otherwise separated segments of the sequence, for example, by chemical synthesis or by manipulation of isolated segments of nucleic acids by genetic engineering techniques. The terms "plasmid", "vector" and "cassette" refer to an extrachromosomal element that usually contains genes that are not part of the cell's central metabolism and usually in the form of circular double-stranded DNA fragments. Such elements may be autonomously replicating sequences, genome integration sequences, phage or nucleotide sequences, linear or circular, of single-stranded or double-stranded DNA or RNA, derived from any source, in which a series of nucleotide sequences is joined or recombined into an original construct that is capable of inserting a promoter fragment and the DNA sequence for a selected gene product along with the appropriate 3' untranslated sequence into a cell. The "transformation cassette" refers to a specific vector that contains an exogenous gene and that has elements other than the exogenous gene that facilitate transformation of a specific host cell. The "expression cassette" refers to a specific vector that contains a gene exogenous and which has elements other than the exogenous gene that allow enhanced expression of that gene in an exogenous host. The terms "recombinant construct", "expression construct", "chimeric construct", "construct" and "recombinant DNA construct" are used interchangeably in the present invention. A recombinant construct comprises an artificial combination of nucleic acid fragments, for example regulatory and coding sequences that are not found together in nature. For example, a chimeric construct may 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 different manner than that found in nature. Such a construct can be used alone or it can be used in conjunction with a vector. If a vector is used, then the choice of vector depends on the method that will be used to transform the host cell as is well known to those skilled in the art. For example, a plasmid vector can be used. The person skilled in the art is well aware of the genetic elements that must be present in the vector in order to successfully transform, select and propagate host cells comprising some of the isolated nucleic acid fragments of the present invention. Those skilled in the art will also recognize the fact that different independent transformation events will result in different levels and patterns of expression (Jones et al., EMBO J. 4:2411-2418 (1985); De Almeida et al., Mol. Gen. Genetics 218:78-86 (1989)) and thus these multiple events must be selected in order to obtain lines that indicate the desired level and pattern of expression. Such selection can be performed by Southern analysis of DNA, Northern analysis of mRNA expression, immunoblotting analysis of protein expression or phenotypic analysis, among others. The term "expression", as used in the present invention, refers to the production of a functional end product (eg, an mRNA or a protein [either precursor or mature]). The term "expression cassette" as used in the present invention refers to a distinct nucleic acid fragment into which a nucleic acid sequence or fragment can be moved. "Mature" protein refers to a post-translationally processed polypeptide (ie, a protein in which any pre- or propeptides present in the primary translation product have been removed). The "precursor" protein refers to the primary translation product of the mRNA (ie, with the pre- and propeptides still present). Pre- and propeptides can be but are not limited to intracellular localization signals. "Stable transformation" refers to the transfer of a nucleic acid fragment into a genome of a host organism, including nuclear and organellar genomes, resulting in genetically stable inheritance. On the other hand, "transient transformation" refers to the transfer of a nucleic acid fragment into the DNA-containing nucleus or organelle of a host organism resulting in gene expression without integration or stable inheritance. Host organisms that contain the transformed nucleic acid fragments are referred to as "transgenic" organisms. "Antisense inhibition" refers to the production of antisense RNA transcripts capable of suppressing target protein expression. "Co-suppression" refers to the production of sense RNA transcripts capable of suppressing the expression of identical or substantially similar exogenous or endogenous genes (US 5,231,020). Co-suppression constructs in plants were previously designed by focusing on overexpression of a nucleic acid sequence that has homology to an endogenous mRNA, in sense orientation, which results in the reduction of all RNA that has homology to the overexpressed sequence (Vaucheret et al., Plant J. 16:651-659 (1998); Gura, Nature 404:804-808 (2000)). The overall efficiency of this phenomenon is low and the extent of RNA reduction is widely variable. Recent work has described the use of "hairpin" structures that incorporate all or part of an mRNA coding sequence in a complementary orientation that results in a "hairpin" structure for the expressed RNA (WO 99 / 53050, published October 21, 1999; WO 02 / 00904, published January 3, 2002). This increases the frequency of co-suppression in recovered transgenic plants. Another variation describes the use of plant viral sequences to direct deletion or "silence" of proximal mRNA coding sequences (WO 98 / 36083, published August 20, 1998). Both co-suppressor phenomena have not been elucidated mechanically, although genetic evidence has begun to resolve this complex situation (Elmayan et al., Plant Cell 10:1747-1757 (1998)). The term "oilseed" refers to those organisms that tend to store their energy source in the form of lipid (Weete, in: Fungal Lipid Biochemistry, 2 a Ed., Plenum, 1980). In general, the cellular oil content of these microorganisms follows a sigmoid curve, in which the lipid concentration increases until it reaches a maximum value in the earlier stationary growth phase or the later log phase and then gradually decreases during the later stationary and late phase. death ( Yongmanitchai and Ward, Appl. Environ. Microbiol. 57:419-25 (1991 )). The term "oil yeast" refers to those microorganisms classified as oil-producing yeasts. It is not uncommon for oleaginous microorganisms to accumulate in excess of about 25% of their cellular dry weight as oil. Examples of oil yeast include, but are not limited to, the following genera: Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon, and Lipomyces. The "Clustal V alignment method" corresponds to the alignment method labeled Clustal V (described by Higgins and Sharp, Cabios. 5:151-153 (1989)) and checked in the Megalign program of the Lasergene bioinformatics computing suite (Dnastar Inc.). , Madison, WI). The "default parameters" are the parameters preset by the manufacturer of the program. For multiple alignments, the parameters correspond to Gap Penalty=10 and Gap Length Penalty=10; and, for pairwise alignments, they are Ktuple 1, Gap Penalty=3, Window=5 and Diagonals Saved=5. After aligning the sequences using the Clustal V program, it is possible to obtain the "percent identity" by viewing the "sequence distances" table in the same program. The present invention relates to an isolated polynucleotide comprising: (a) a nucleotide sequence encoding a polypeptide having delta-8 desaturase activity, the polypeptide having an amino acid sequence consisting essentially of SEQ ID NO: 2 or 113; or (b) a complement of the nucleotide sequence, where the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary. This delta-8 desaturase can be used alone or in combination with other desaturase and elongase components to produce various omega-6 and omega-3 PUFAs including, for example, DGLA, ETA, ARA, EPA, DPA and / or DHA ( Figure 9). One skilled in the art will recognize appropriate combinations of the delta-8 desaturase of the present invention together with a delta-5 desaturase, a delta-6 desaturase, a delta-12 desaturase, a delta-15 desaturase, a delta-17 desaturase, a delta-9 desaturase, one delta-9 elongase, one elongase C 14 / ci 6 , an elongase, a C18 / C20 elongase and / or a C20 / C22 elongase, based on on the specific host cell (and its native PUFA profile and / or desaturase and / or elongase profile), substrate availability, and desired end product(s). In another embodiment, the present invention relates to a recombinant construct comprising the polynucleotide of the present invention operably linked to at least one regulatory sequence. Systems, Cassettes and Plant Expression Vectors As noted above, a promoter is a DNA sequence that directs the cellular machinery of a plant to produce RNA from the coding sequence contiguous downstream (3') of the promoter. The promoter region influences the rate, stage of development, and cell type in which the RNA transcript of the gene is made. The transcribed RNA is processed to produce the mRNA that serves as a template strand for the translation of the RNA sequence into the amino acid sequence of the encoded polypeptide. The 5' untranslated leader sequence is a region of mRNA upstream of the protein coding region that may play a role in mRNA initiation and translation. The 3' transcription termination / polyadenylation signal is an untranslated region downstream of the protein coding region that functions in the plant cell to cause termination of transcribed RNA and the addition of polyadenylate nucleotides to the 3' end of the RNA. The origin of the promoter selected to drive expression of the coding sequence is not important as long as it has sufficient transcriptional activity to carry out the present invention by expressing mRNA translatable to the desired nucleic acid fragments in the desired host tissue at the right time. Heterologous or non-heterologous (i.e., endogenous) promoters can be used to practice the present invention. For example, appropriate promoters include, but are not limited to, the major alpha subunit of the beta conglycinin promoter, Kunitz inhibitor trypsin 3 promoter, annexin promoter, Gly1 promoter, conglycinin beta promoter beta subunit, P34 / Gly Bd m 30K promoter, albumin promoter, LegA1 promoter and LegA2 promoter. The annexin promoter, or P34, is described in WO 2004 / 071178 (published August 26, 2004). The activity level of the annexin promoter is comparable to that of many known strong promoters, such as: (1) the CaMV 35S promoter ( Atanassova et al., Plant Mol. Biol. 37:275-285 (1998 ); Battraw and Hall , Plant Mol. Biol. 15:527-538 (1990), Holtorf et al, Plant Mol. Biol., 29:637-646 (1995), Jefferson et al., Embo J. 6:3901-3907 (1987); Wilmink et al., Plant Mol. Biol. 28:949-955 (1995)); (2) the Arabidopsis oleosin promoters (Plant et al., Plant Mol. Biol. 25:193-205 (1994); Li, Texas A&M University Ph.D. dissertation, pp. 107-128 (1997) )); (3) the Arabidopsis ubiquitin extension protein promoters ( Callis et al., J. BiolChem. 265(21):12486-93 (1990 )); (4) a promoter from the tomato ubiquitin gene ( Rollfinke et al, Gene. 211(2):267-76 (1998 )); (5) a soy heat shock protein promoter (Schoffl et al., Mol Gen Genet. 217(2-3):246-53 (1989)); and, (6) a maize histone H3 gene promoter ( Atanassova et al., Plant Mol. Biol. 37(2):275-85 (1989 )). Another useful feature of the annexin promoter is its expression profile in developing seeds. The annexin promoter is most active in developing seeds at early stages (before 10 days after pollination) and is largely inactive at later stages. The expression profile of the annexin promoter is different from that of many seed-specific promoters, for example the seed storage protein promoters, which often provide higher activity at later stages of development (Chen et al., Dev. Genet 10:112-122 (1989); Ellerstrom et al., Plant Mol. Biol. 32:1019-1027 (1996); Keddie et al., Plant Mol. Biol. 24:327-340 (1994); Plant et al., (above); Li, (above)). The annexin promoter has a more conventional expression profile but remains distinct from other known seed-specific promoters. Thus, the annexin promoter will be a very attractive candidate when overexpression or deletion of a gene in embryos is desired at an early developmental stage. For example, it may be desirable to overexpress a gene that regulates early embryo development or a gene involved in metabolism prior to seed maturation. After identification of an appropriate promoter for expression of a specific coding sequence, the promoter is then operably turned on in a sense orientation using conventional means well known to those skilled in the art. Standard recombinant DNA and molecular cloning techniques used in the present invention are well known in the art and described by Sambrook, J., Fritsch, E.F., and Maniatis, T., Molecular Cloning: A Laboratory Manual, 2 a ed., Cold Spring Harbor Laboratory: Cold Spring Harbor, NY (1989) (hereinafter "Maniatis"); by Silhavy, T.J., Bennan, M.L. and Enquist, L.W., Experiments with Gene Fusions, Cold Spring Harbor Laboratory: Cold Spring Harbor, NY (1984); and by Ausubel, F.M. et al., Current Protocols in Molecular Biology, published by Greene Publishing Assoc, and Wiley-Interscience (1987). Plant Transformation Once the recombinant construct is obtained, it can then be inserted into a reiv 6 plant cell of choice by methods well known to those skilled in the art (eg, transfection, transformation and electroporation). Oilseed plant cells are the preferred plant cells. The transformed plant cell is then cultured and regenerated under appropriate conditions that allow expression of PUFA from long chain which is then optionally recovered and purified. The recombinant constructs of the present invention can be inserted into a plant cell; or, alternatively, each construct can be inserted into separate plant cells. Expression in a plant cell can be carried out in a transient or stable manner as described above. The desired long chain PUFAs can be expressed in seed. Also within the scope of the present invention are encompassed seeds or plant parts obtained from such transformed plants. Plant parts include differentiated and undifferentiated tissues including, but not limited to: roots, stems, shoots, leaves, pollen, seeds, tumor tissue, and various forms of cells and culture (e.g., single cells, protoplasts , embryos and callus tissue). The plant tissue can be in the plant or in a plant organ, in a tissue or in a cell culture. The term "plant organ" refers to the plant tissue or group of tissues that constitute the morphologically and functionally distinct part of a plant. The term "genome" refers to the following: (1) The entire complement of genetic material (genes and non-coding sequences) is present in every cell of an organism or virus or organelle. (2) A complete set of chromosomes inherited as the (haploid) unit from a parent. Thus, the present invention also relates to a method for transforming a cell, which comprises transforming a cell with the recombinant construct of the present invention and selecting these transformed cells with the recombinant construct according to claim 4. Also of interest is a method for producing a transformed plant, which comprises transforming a plant cell with the polynucleotide of the present invention and regenerating a plant from the transformed plant cell. Methods for transforming dicots (mainly by using Agrobacterium tumefaciens) and obtaining transgenic plants have been published, among others, for: cotton (US 5,004,863; US 5,159,135); soy (US 5,569,834; US 5,416,011); Brassica (US 5,463,174); peanut (Cheng et al., Plant Cell Rep. 15:653-657 (1996); McKently et al., Plant Cell Rep. 14:699-703 (1995)); papaya (Ling, K. et al., Biotechnology 9:752-758 (1991)); and pea ( Grant et al., Plant Cell Rep. 15:254-258 (1995 )). For a review of other commonly used plant transformation methods, see Newell, C.A. ( Mol. Biotechnol. 16:53-65 (2000 )). One of these transformation methods generally uses Agrobacterium rhizogenes (Tepfler, M. and Casse-Delbart, F. Microbiol. Sci. 4:24-28 (1987)). Transformation of soybeans using direct application of DNA has been published using PEG fusion (WO 92 / 17598), electroporation (Chowrira, G.M. et al., Mol. Biotechnol. 3:17-23 (1995); Christou, P. et al., Proc. Natl. Acad. Sci. USA. 84:3962-3966 (1987)), microinjection or particle bombardment (McCabe, D.E. et. al., Bio / Technology 6:923 (1988); Christou et al., Plant Physiol., 87:671-674 (1988)). There are a variety of methods for regenerating plants from plant tissue. The specific regeneration method will depend on the starting plant tissue and the specific plant species to be regenerated. The regeneration, growth and cultivation of plants from single plant protoplast transformants or from multiple transformed explants is well known in the art (Weissbach and Weissbach, in: Methods for Plant Molecular Biology, (Eds.), Academic : San Diego, CA (1988)). This regeneration and growth process typically includes the steps of selecting transformed cells and growing individualized cells through the usual stages of embryonic development through the rooted seedling stage. Embryos and transgenic seeds are similarly regenerated. The resulting transgenic rooted shoots are then planted in an appropriate plant growing medium such as soil. Preferably, the regenerated plants are self-pollinated to provide homozygous transgenic plants. If not, the pollen obtained from the regenerated plants is crossed to plants grown by seeds of agronomically important lines. On the other hand, pollen from plants of these important lineages is used to pollinate the regenerated plants. A transgenic plant of the present invention that contains a desired polypeptide is grown using methods well known to those skilled in the art. In addition to the procedures discussed above, those skilled in the art are familiar with standard resource materials that describe specific conditions and procedures for the construction, manipulation, and isolation of macromolecules (e.g., DNA molecules, plasmids, etc.), generation of recombinant DNA fragments, recombinant expression constructs, and the selection and isolation of clones. See, for example: Sambrook et al., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor: NY (1989); Maliga et al., Methods in Plant Molecular Biology, Cold Spring Harbor: NY (1995); Birren et al., Genome Analysis: Detecting Genes, Vol. 1, Cold Spring Harbor: NY (1998); Birren et al., Genome Analysis: Analyzing DNA, Vol. 2, Cold Spring Harbor: NY (1998); Plant Molecular Biology: A Laboratory Manual, eds. Clark, Springer: NY (1997). Examples of oilseed plants include, but are not limited to, soybean, Brassica species, sunflower, corn, cotton, linseed, safflower. Examples of polyunsaturated fatty acids that have at least 20 carbon atoms and 5 or more carbon-carbon double bonds include, but are not limited to, omega-3 fatty acids such as EPA, DPA, and DHA. Seeds obtained from such plants are also within the scope of the present invention, as well as oil obtained from such seeds. In one embodiment, the present invention relates to an oilseed plant comprising: (a) a first recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide operably linked to at least a regulatory sequence; and (b) at least one additional recombinant DNA construct comprising an isolated polynucleotide operably linked to at least one regulatory sequence encoding a polypeptide selected from the group consisting of delta-4, delta-5, delta -6, delta-9, delta-12, delta-15 and delta-17 desaturase, delta-9 elongase, elongase Cis to C22 and elongase C 2 o to C 24 . Such desaturases are discussed in US Patents 6,075,183, US 5,968,809, US 6,136,574, US 5,972,664, US 6,051,754, US 6,410,288 and WO 98 / 46763, WO 98 / 46764, WO 00 / 12720, WO 00 / 40705. The choice of combination of cassettes used depends in part on the desaturase profile and / or the PUFA profile of the oilseed plant cells to be transformed and the LC-PUFA that must be expressed. In another aspect, the present invention relates to a method of manufacturing long-chain polyunsaturated fatty acids in a plant cell, which comprises: (a) transforming a cell with the recombinant construct of the present invention; and (b) the selection of those transformed cells that form the long-chain polyunsaturated fatty acids. In yet another aspect, the present invention relates to a method for producing at least one polyunsaturated fatty acid in a soybean cell, which comprises: (a) transforming a soybean cell with a first recombinant DNA construct comprising an isolated polynucleotide encoding a delta-8 desaturase polypeptide operably linked to at least one regulatory sequence and to at least one additional recombinant construct of DNA comprising an isolated polynucleotide operably linked to at least one regulatory sequence encoding a polypeptide selected from the group consisting of delta-4, delta-5, delta-6, delta-9, delta-12 , delta-15 and delta-17 desaturase, delta-9 elongase, elongase Ci 8 to C22 and elongase C 2 o to C24; (b) regenerating a soybean plant from the transformed cell of step (a); and (c) selecting seeds obtained from plants in step (b) that have an altered level of polyunsaturated fatty acids when compared to the level in seeds obtained from an untransformed soybean plant. Plant Seed Oils: Isolation and Hydrogenation Methods for isolating seed oils are well known in the art: (Young et al., Processing of Fats and Oils, in The Lipid Handbook, Gunstone et al., eds., chapter 5, pages 253 to 257; Chapman & Hall: London (1994)). For example, soybean oil is produced using a series of steps that involve extracting and purifying an edible oil product from the seed that carries the oil. Soybean oils and soy by-products are produced using the generalized steps shown in the table below. Table 3 Generalized Steps for the Production of Soybean Oil and Byproducts Process Step Process Impurities removed and / or by-products obtained #1 Soybean #2 Oil extraction Flour #3 Lecithin degumming #4 Alkaline or physical refinement Gums, free fatty acids, pigments #5 Water washing Soap #6 Color bleach , soap, metal #7 (Hydrogenation) #8 (Wintering) Stearin #9 Deodorization Free fatty acids, tocopherols, sterols, volatiles #10 Oil products In general, soybean oil is produced by employing a series of steps that involve extracting and purifying an edible oil product from the oil-bearing seed. Soybean oils and soy by-products are produced using the generalized steps shown in the diagram below. Process Impurities Removed / Obtained by-products soybean seed I Flour Oil Extraction Degumming -+ Lecithin Alkaline or physical refining -» Gums, Free Fatty Acids, Pigments Washing with Water -> Soap Bleach -> Color, Soap, Metal (Hydrogenation) >L (Wintering) -> Stearin FFA Deodorization, Tocopherols, Sterols, Volatiles Oil Products More specifically, soybean seeds are cleaned, mixed, hulled and hulled, thereby increasing the efficiency of oil extraction. Oil extraction is usually accomplished by solvent extraction (eg hexane) but can also be achieved by a combination of solvent extraction and / or physical pressure. The resulting oil is called crude oil. Crude oil can be degummed by hydration of phospholipids and other polar and neutral complexes that facilitate its separation from the non-hydrated triglyceride fraction (soybean oil). The resulting lecithin gums can be further processed to manufacture the commercially important lecithin products used in a variety of food and industrial products as emulsifiers and release agents (i.e. anti-adherents). Degummed oil can be further refined to remove impurities (mainly free fatty acids, pigments and residual gums). the refinement It is accomplished by adding a caustic agent that reacts with the free fatty acid to form soap and hydrate the phosphatides and proteins in the crude oil. Water is used to wash away the soap residue formed during refining. The sludge by-product (soapstock) can be used directly in animal feed or it can be acidified to recover free fatty acids. The color is removed by adsorption with a bleaching earth which removes most of the chlorophyll and carotenoid compounds. Refined oil can be hydrogenated, thereby 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 refrigeration conditions. Deodorization (mainly through vacuum steam distillation) is the last step and is designed to remove compounds that impart odor or flavor to the oil. Other valuable by-products such as tocopherols and sterols can be removed during the deodorization process. The deodorized distillate containing these by-products can be sold for the production of natural vitamin E and other high value pharmaceuticals. Refined, bleached, (hydrogenated, fractionated) and deodorized oils and fats can be packaged and sold directly or further processed into more specialized products. A more detailed reference to soybean seed processing, soybean oil production, and by-product utilization can be found in Erickson, Practical Handbook of Soybean Processing and Utilization, The American Oil Chemists’ Society and United Soybean Board (1995). Soybean oil is liquid at room temperature because it has a relatively low content of saturated fatty acids when compared to oils such as coconut, palm, palm kernel and cocoa butter. Many processed fats (including spreadables, confectionery fats, solid butters, margarines, cooking butter, etc.) vary in fastness at room temperature and can only be produced from soybean oil by altering its physical properties. This is most commonly achieved through catalytic hydrogenation. Hydrogenation is a chemical reaction in which hydrogen is added to unsaturated fatty acid double bonds with the aid of a catalyst such as nickel. High oleic soybean oil contains oleic, LA and linolenic unsaturated fatty acids, each of which can be hydrogenated. Hydrogenation has two primary effects. First, the oxidative stability of the oil is increased as a result of the reduction in the unsaturated fatty acid content. Second, the physical properties of the oil are altered due to fatty acid modifications which raise the melting point resulting in a semi-liquid or solid fat at room temperature. There are many variables that affect the hydrogenation reaction, which in turn change the composition of the final product. Operating conditions including pressure, temperature, catalyst type and concentration, agitation, and reactor design are among the most important parameters that can be controlled. Selective hydrogenation conditions can be used to hydrogenate more unsaturated fatty acids instead of less unsaturated ones. Very light or rapid hydrogenation is often employed to increase the stability of liquid oils. Additional hydrogenation converts a liquid oil into a physically solid fat. The degree of hydrogenation depends on the desired melting and performance characteristics designed for the specific end product. Liquid vegetable fats (used in the manufacture of baked goods, solid fats and vegetable fats used for commercial frying and baking) and base stocks for making margarine are among a myriad of possible fat and oil products obtained through hydrogenation. A more detailed description of hydrogenation and products hydrogenates can be found in Patterson, H.B.W.; Hydrogenation of Fats and Oils: Theory and Practice. The American Oil Chemists' Society (1994). Hydrogenated oils have also become controversial due to the presence of trans fatty acid isomers that result from the hydrogenation process. Intake of large amounts of trans isomers has been linked to harmful health effects including increased ratios between low-density lipoproteins and high-density lipoproteins in blood plasma and increased risk of coronary heart disease. Compared to other vegetable oils, the oils of the present invention are believed to function similarly to other oils in food applications from a physical point of view. Partially hydrogenated oils, such as soybean oil, are used extensively as ingredients for smooth spreads, margarine, and shortening for baking and frying. Examples of food products or food analogues into which modified seed oils or modified seeds of the present invention may be incorporated include a meat product such as a processed meat product, a cereal food product, a snack food product, a baked product , a fried food product, a health food product, an infant formula, a beverage, a nutritional supplement, a dairy product, a pet food product, an animal feed or an aquaculture food product. Food analogues can be obtained by employing processes known to those skilled in the art. US Patents 6,355,296 B1 and US 6,187,367 B1 describe emulsified meat analogues and emulsified meat extenders. US Patent 5,206,050 B1 describes a soy protein curd useful for cooked food analogues (it can also be used as a process to form a curd useful in the manufacture of food analogues). US Patent 4,284,656 to Hwa describes a Useful soy protein curd for food analogues. US Patent 3,988,485 to Hibbert et al. describes a meat-like protein food formed from interwoven vegetable protein fibers. US Patent 3,950,564 to Puski et al. describes a process for making a soy based meat substitute and US Patent 3,925,566 to Reinhart et al. describes a simulated meat product. For example, soy protein that has been processed to impart a structure, chunk, or fiber for use as a food ingredient is called "textured soy protein" (TSP). PTSs are typically manufactured to resemble meat, seafood or poultry in structure and appearance when hydrated. Mention may be made of meat analogues, cheese analogues, milk analogues and the like. Meat analogues made from soy contain soy protein or tofu and other ingredients mixed together to simulate various types of meat. These meat alternatives are sold as frozen, canned or dried foods. Generally, they can be used in the same way as the foods they replace. Meat alternatives made from soy are excellent sources of protein, iron and B vitamins. Examples of meat analogues include, but are not limited to, ham analogues, sausage analogues, bacon analogues and the like. Food analogues can be classified as an imitation or substitutes depending on their functional and compositional characteristics. For example, an imitation cheese need only resemble the cheese it is designed to replace. However, a product can generally be called a substitute cheese only if it is nutritionally equivalent to the cheese it is replacing and meets the minimum compositional requirements for that cheese. In this way, the cheese substitute will often have higher levels of protein than cheese imitations and will be fortified with vitamins and minerals. Milk analogues or non-dairy food products include, but are not limited to, milk analogues, non-dairy frozen desserts such as those made from soy and / or soy protein products. Meat products cover a wide variety of products. In the United States, "meat" includes "red meat" produced from beef cattle, pigs, and sheep. In addition to red meat, there are poultry items that include chickens, turkeys, geese, guinea fowl, ducks, fish and shellfish. There is a wide variety of seasoned and processed meat products: fresh, cured and fried, and cured and baked. Sausages and sausages are examples of processed meat products. Accordingly, the term "meat products" as used in the present invention includes, but is not limited to, processed meat products. A cereal food product is a food product derived from the processing of a cereal grain. A cereal grain includes any plant in the grass family that yields an edible grain (seed). The most popular grains are barley, corn, millet, oats, quinoa, rice, rye, sorghum, triticale, wheat and wild rice. Examples of a cereal food product include, but are not limited to, whole grain, crushed grain, granulated cereal, flour, bran, germ, breakfast cereals, extruded foods, pasta and the like. A baked product comprises some of the cereal food products mentioned above and cooked or processed in a manner comparable to baking, i.e. dried or hardened by subjecting it to heat. Examples of a baked product include, but are not limited to, breads, cakes, donuts, breadcrumbs, baked snacks, mini savory crackers, mini sweet crackers, mini muffins and mini pretzels. As mentioned above, the oils of the present invention can be used as an ingredient. A snack food product comprises any of the food products described above or below. A fried food product comprises any of the food products described above or below that has been fried. A health food product is any food product that provides a health benefit. Many food products derived from oilseeds can be considered as health foods. Beverages can be in a liquid form or in a dry powder form. For example, non-carbonated drinks can be mentioned; fresh, frozen, canned or concentrated fruit juices; natural or flavored dairy drinks, etc. Nutritious infant and adult formulas are well known in the art and are commercially available (eg, Similac®, Ensure®, Jevity® and Alimentum® from Ross Products Division, Abbott Laboratories). Infant formulas are in the form of liquids or reconstituted dry powders given to infants and young children. They serve as substitutes for human milk. Infant formulas play a special role in babies' diets as they are often the only source of nutrients for babies. While breastfeeding is still the best nutrition for babies, infant formula also ensures that babies not only survive but also grow. Infant formula is becoming closer and closer to breast milk. A dairy product is a product derived from milk. a product milk analogue or non-dairy is derived from a source other than milk, for example soy milk as discussed above. These products include, but are not limited to, whole milk, skimmed milk, fermented milk products such as yogurt or curd, cream, butter, condensed milk, dehydrated milk, coffee whitener, coffee creamer, ice cream, cheese, etc. A pet food product is a product that lends itself to feeding a pet such as a dog, cat, bird, reptile, fish, rodent and the like. These products may include the above cereal products and health food products, such as meat and meat by-products, soy protein products, grass and hay, including but not limited to alfalfa, phelloli, oats or barley grass, vegetables and the like. An animal feed is a product that lends itself to feeding animals such as turkeys, chickens, cattle, pigs and the like. Like the pet foods above, these products may include cereal products and health food products, soy protein products, meat and meat by-products, and grass and hay products as listed above. Aquaculture feed is a product that lends itself to be used for in-water farming that involves the propagation, cultivation or rearing of aquatic organisms, animals and / or plants in fresh or salt water. Microbial Biosynthesis of Fatty Acids The process of synthesizing de novo palmitate (16:0) in oleaginous microorganisms is described in WO 2004 / 101757. This fatty acid is the precursor of the longer chain saturated and unsaturated fatty acid derivatives, which are formed through the action of elongases and desaturases. For example, palmitate is converted to its unsaturated derivative [palmitoleic acid (16:1)] by the action of a delta-9 desaturase; similarly, palmitate is elongated to form stearic acid (18:0), which can be converted to its unsaturated derivative by a delta-9 desaturase to thereby yield oleic acid (18:1). Triacylglycerols (the primary storage unit for fatty acids) are formed by the esterification of two acyl-CoA molecules to glycerol-3-phosphate to yield 1,2-diacylglycerolphosphate (generally identified as phosphatidic acid). The phosphate is then removed by phosphatidic acid phosphatase to give 1,2-diacylglycerol. Triacylglycerol is formed upon the addition of a third fatty acid by the action of an acyl-diacylglycerol transferase. Genes Involved in Omega Fatty Acid Production Many microorganisms, including algae, bacteria, fungi and yeast, can synthesize PUFAs and omega fatty acids in the ordinary course of cellular metabolism. Fungi including Schizochytrium aggregatum, a species of the genus Thraustochytrium, and Morteriella alpina are particularly well studied. Additionally, many dinoflagellates (Dinophyceae) naturally produce high concentrations of PUFAs. Thus, a variety of genes involved in oil production have been identified through genetic means and the DNA sequences of some of these genes are publicly available. See, for example: AY131238, Y055118, AY055117, AF296076, AF007561, L11421, NM_031344, AF465283, AF465281, AF110510, AF465282, AF419296, AB052086, AJ250735, 9, AF126-86 deletions); AF199596, AF226273, AF320509, AB072976, AF489588, AJ510244, AF419297, AF07879, AF067654, AB022097 (delta-5 desaturases); AAG36933, AF110509, AB020033, AAL13300, AF417244, AF161219, AY332747, AAG36933, AF110509, AB020033, AAL13300, AF417244, AF161219, X86736, AF240777, AB0076540 AP002063 (delta-12 desaturases); NP_441622, BAA18302, BAA02924, AAL36934 (delta-15 desaturases); AF338466, AF438199, E11368, E11367, D83185, U90417, AF085500, AY504633, NM_069854, AF230693 (delta-9 desaturases); AF390174 (delta-9 elongase); and AX464731, NM_119617, NM_134255, NM_134383, NM_134382, NM_068396, NM_068392, NM 070713, NM_068746, NM_064685 (ellongases). In addition, the patent literature provides many additional DNA sequences of genes (and / or details regarding several of the above genes and their isolation methods) involved in PUFA production [e.g., US 5,968,809 (delta-6 desaturases ); US 5,972,664 and US 6,075,183 (delta-5 desaturases); WO 94 / 11516, US 5,443,974 and WO 03 / 099216 (delta-12 desaturases); WO 93 / 11245 (delta-15 desaturases); WO 91 / 13972 and US 5,057,419 (delta-9 desaturases); US 2003 / 0196217 A1 (delta-17 desaturase); and WO 00 / 12720, WO 2002 / 077213 and US 2002 / 0139974 A1 (elongases)]. As should be obvious to those skilled in the art, the particular functionalities required for introduction into a host microbial organism to produce a specific PUFA end product will depend on the host cell (and its native PUFA profile and / or its native PUFA profile). / elongase), substrate availability, and desired final product(s). LA, GLA, EDA, DGLA, ARA, ALA, STA, ETrA, ETA, EPA, DPA and DHA can be produced in oleaginous yeasts by introducing various combinations of the following PUFA enzyme functionalities: a delta-4 desaturase, a delta -5 desaturase, one delta-6 desaturase, one delta-8 desaturase, one delta-12 desaturase, one delta-15 desaturase, one delta-17 desaturase, one delta-9 desaturase, one C14 / C elongase 16 , a Ci6 / Ci elongase 8 , a Cis / C elongase 2 o and / or an elongase C 20 / Ç 22 . the person skilled in the art will be able to identify the various candidate genes that encode each of the above enzymes, according to publicly available literature (e.g., GenBank), patent literature, and experimental analysis of microorganisms that have the ability to produce PUFAs. The sequences can be derived from any source, for example, isolated from a natural source (bacteria, algae, fungi, plants, animals, etc.), produced through a semi-synthetic route, or synthesized de novo. In some embodiments, manipulation of genes endogenous to the host is preferred; for other purposes, insertion of heterologous genes is required. Although the specific source of the desaturase and elongase genes inserted in the host is not essential to the present invention, considerations for choosing a specific polypeptide that has a desaturase or elongase activity include: (1) the substrate specificity of the polypeptide; (2) whether the polypeptide or a component thereof is a rate-limiting enzyme; (3) whether the desaturase or elongase is essential for the synthesis of a desired PUFA; and / or (4) the cofactors required by the polypeptide. The expressed polypeptide preferably has parameters compatible with the biochemical environment of its position in the host cell. For example, the polypeptide may have to compete for substrate with other enzymes in the host cell. Km and polypeptide-specific activity analyzes are therefore taken into account in determining the suitability of a given polypeptide to modify PUFA production in a given host cell. The polypeptide used in a specific host cell is one that can function under the biochemical conditions present in the intended host cell, but otherwise can be any polypeptide that has a desaturase or elongase activity capable of modifying the desired PUFA. In some cases, the host organism in which it is desirable to production of PUFAs will have the endogenous genes that encode the enzymes of the biosynthetic pathway of some PUFA. For example, oil yeasts can typically produce 18:2 fatty acids (and some have the additional ability to synthesize 18:3 fatty acids); thus, oil yeasts typically have native delta-12 desaturase activity and may also have delta-15 desaturases. In some embodiments, therefore, expression of the native desaturase enzyme is preferred over a heterologous enzyme or ("exogenous") enzyme since: (1) the native enzyme is optimized for interaction with other enzymes and proteins within the cell; and (2) heterologous genes are unlikely to share the same codon preference in the host organism. Additionally, advantages are incurred when the native gene sequence is known, as it allows easy disruption of the endogenous gene by targeted disruption. In many cases, however, the appropriate desaturases and elongases are not present in the host organism of choice to allow production of the desired PUFA products. Thus, the introduction of heterologous genes is necessary. In one embodiment of the present invention, work was carried out with the aim of developing an oleaginous yeast that accumulates oils rich in omega-6 and / or long-chain omega-3 fatty acids. In order to express the genes encoding the delta-9 elongase / delta-8 desaturase pathway for ARA and EPA biosynthesis in these organisms, it was therefore necessary to: (1) identify an appropriate desaturase that functions relatively efficiently in yeast oilseed based on substrate feeding trials; and, (2) subjecting the desaturase gene to codon optimization techniques (below) to further enhance expression of the heterologous enzymes in the alternative oilseed yeast host, thereby allowing maximum production of omega-3 fatty acids and / or omega-6. Optimization of Omega Fatty Acid Genes for Expression in Specific Organisms While the specific source of a PUFA desaturase or elongase is not essential in the present invention, it will be obvious to one skilled in the art that the heterologous genes will be expressed with varying efficiencies in an alternative host. Thus, production of omega-3 and / or omega-6 PUFAs can be optimized by selecting a specific desaturase or elongase whose expression level in a heterologous host is preferred over expression of an alternative desaturase or elongase. in the host organism of interest. Furthermore, it may be desirable to modify the expression of PUFA-specific biosynthetic pathway enzymes to achieve a more favorable efficiency of conversion of each, according to the composition of the specific PUFA product of interest. A variety of genetic engineering techniques are available to optimize the expression of a specific enzyme. Two techniques include codon optimization and gene mutation, as described below. Genes produced, for example, by either of these two methods, which have a desaturase and / or elongase activity would be useful in the present invention for the synthesis of omega-3 and / or omega-6 PUFAs. Codon Optimization As will be appreciated by one skilled in the art, it is often useful to modify a portion of codons encoding a specific polypeptide that must be expressed in an exogenous host, so that the modified polypeptide uses the codons that are preferred by the alternative host. The use of host-preferred codons can substantially enhance expression of exogenous genes encoding the polypeptide. In general, host-preferred codons can be determined within a specific host species of interest by examining codon usage in proteins (preferably those expressed in the greatest amount) and by determining which codons are used with the highest frequency. Thus, the coding sequence for a polypeptide of interest that has desaturase or elongase activity can be synthesized in whole or in part using the codons preferred in the host species. The entire DNA (or portions) can also be synthesized to remove all destabilizing sequences or regions of secondary structure that are present in the mRNA transcript. All DNA (or portions) can also be synthesized to change the base composition to a preferable composition in the desired host cell. In the present invention, it was desirable to modify a portion of the codons encoding the polypeptide having a delta-8 desaturase activity, to enhance expression of the gene in the oleaginous yeast of Yarrowia lipolytica. The nucleic acid sequence of the native gene (eg, the Euglena gracilis delta-8 desaturase defined in the present invention as Eg5) has been modified to employ host-preferred codons. This wild-type desaturase is 421 amino acids (SEQ ID NO:2); in the codon-optimized gene created in the present invention (SEQ ID NO:112), 207 bp of the 1263 bp coding region (which corresponds to 192 codons) codon-optimized and the translation initiation site was modified. Those skilled in the art will appreciate that this optimization method will be equally applicable to other genes in the omega-3 / omega-6 fatty acid biosynthetic pathway (see, for example, WO 2004 / 101753, incorporated in the present invention entirely by way of reference). Furthermore, modulation of E. gracilis delta-8 desaturase is just one example; numerous other heterologous delta-8 desaturases from varying sources can be codon-optimized to improve their expression in an oilseed yeast host. THE The present invention comprises the complete sequences of the synthetic codon-optimized gene as reported in the attached Sequence Listing, the complement of those complete sequences, and substantial portions of those sequences. Gene Mutation The methods for synthesizing the sequences and joining them are well established in the literature. For example, in in vitro mutagenesis and selection, site-directed mutagenesis, error-prone PCR (Melnikov et al., Nucleic Acids Research, 27(4):1056-1062 (Feb. 5, 1999)), the " gene rearrangement" or other means may be employed to obtain mutations of naturally occurring desaturase or elongase genes (such mutations may include deletions, insertions, and point mutations or combinations thereof). This would allow the production of a polypeptide that has a desaturase or elongase activity, respectively, in vivo, with more desirable physical and kinetic parameters for function in the host cell such as a longer half-life or a higher production rate of a desired PUFA. Or, if desired, the regions of a polypeptide of interest (ie, a desaturase or an elongase) important for enzymatic activity can be determined through routine mutagenesis, expression of the resulting mutant polypeptides, and determination of their activities. An overview of these techniques is described in WO 2004 / 101757. All such mutant proteins and the nucleotide sequences encoding them which are derived from the codon-optimized gene described in the present invention are within the scope of the present invention. Microbial Production of Omega-3 and / or Omega-6 Fatty Acids Microbial production of omega-3 and / or omega-6 fatty acids has several advantages. For example: (1) many microbes are known with extremely simplified oil compositions compared to those of higher organisms, making purification of desired components easier; (2) microbial production is not subject to fluctuations caused by external variables such as weather and food supply; (3) microbially produced oil is substantially free of contamination by environmental pollutants; (4) microbes can provide PUFAs in particular forms that can have specific uses; and (5) microbial oil production can be manipulated by controlling culture conditions, primarily by providing particular substrates for microbially expressed enzymes or by adding compounds / genetic engineering to suppress unwanted biochemical pathways. In addition to these advantages, the production of omega-3 and / or omega-6 fatty acids from recombinant microbes provides the ability to alter the naturally occurring microbial fatty acid profile by providing new biosynthetic pathways in the host or by suppressing unwanted pathways. , thereby increasing levels of desired PUFAs or conjugated forms thereof and decreasing levels of unwanted PUFAs. For example, it is possible to modify the ratio of omega-3 and omega-6 fatty acids produced in this way to exclusively produce omega-3 or omega-6 fatty acids by eliminating alternative omega fatty acid production or engineering production of omega-6 fatty acids. a specific PUFA without significant accumulation of other PUFA products downstream or upstream (e.g., allowing the biosynthesis of ARA, EPA and / or DHA via the delta-9 elongase / delta-8 desaturase pathway, thereby preventing the synthesis the GLA and / or the STA). Microbial Systems, Cassettes and Expression Vectors The genes and gene products described in the present invention can be produced in heterologous microbial host cells, particularly in oleaginous yeast cells (eg Yarrowia lipolytica). Expression in recombinant microbial hosts may be useful for the production of various PUFA pathway intermediates or for the modulation of PUFA pathways that already exist in the host for the synthesis of new products that so far are not possible using the host. Microbial expression systems and expression vectors that contain regulatory sequences that direct high-level expression of exogenous proteins are well known to those skilled in the art. Any of these can be used to construct chimeric genes for the production of any of the preferred elongase and / or desaturase sequence gene products. These chimeric genes can then be inserted into the appropriate microorganisms through transformation to provide high level expression of the encoded enzymes. Consequently, the introduction of chimeric genes encoding a PUFA biosynthetic pathway under the control of appropriate promoters is expected to result in increased production of omega-3 and / or omega-6 fatty acids. It is contemplated that it is useful to express various combinations of desaturase and elongase genes from these PUFAs together in a host microorganism. It will be obvious to those skilled in the art that the particular genes included within a particular expression cassette will depend on the host cell, its ability to synthesize PUFAs using native desaturases and elongases, the availability of the substrate, and the final product(s) (is) desired. For example, it may be desirable for an expression cassette to be constructed which comprises genes encoding one or more of the following enzymatic activities: a delta-4 desaturase, a delta-5 desaturase, a delta-6 desaturase, a delta- 8 desaturase, a delta-12 desaturase, a delta-15 desaturase, a delta-17 desaturase, a delta-9 desaturase, a C14 / C16 elongase, a C16 / C18 elongase, a C18 / C20 elongase and / or a 620 / 622 elongase. comprises exposing a fatty acid substrate to the PUFA enzyme(s) described in the present invention, in such a manner that the substrate is converted to the desired fatty acid product. In this way, each PUFA gene and each corresponding enzyme product described in the present invention (e.g., a wild-type, codon-optimized enzyme, synthetic enzyme, and / or mutant that has an appropriate desaturase or elongase activity) can be used directly or indirectly for the production of PUFAs. Direct production of PUFAs occurs when the fatty acid substrate is converted directly into the desired fatty acid product without any intermediate steps or intermediate pathways. For example, ARA production would occur in a host cell that produces or supplies DGLA, by adding or introducing into said cell an expression cassette that provides delta-5 desaturase activity. Similarly, the expression of the delta-8 desaturase of the present invention allows the direct synthesis of DGLA and ETA (when EDA and ETrA, respectively, are provided as substrate). Thus, for example, the present invention relates to a production method of DGLA or ETA, respectively, which comprises: (a) employing an oleaginous yeast comprising: (i) a gene encoding a delta-8 desaturase polypeptide as set forth in SEQ ID NO:112; and (ii) a source of desaturase substrate consisting of EDA or ETrA, respectively; and, (b) growing the yeast from step (a) in the presence of an appropriate fermentable carbon source in which the gene encoding a delta-8 desaturase polypeptide is expressed and EDA is converted to DGLA or ETrA is converted to ETA, respectively; and, (c) optionally recovering the DGLA or ETA, respectively, from step (b). On the other hand, the multiple genes encoding the PUFA biosynthetic pathway can be used in combination so that a series of reactions occur to produce a desired PUFA. For example, the expression cassette(s) encoding an elongase, delta-5 desaturase, delta-17 desaturase, and delta-4 desaturase activity must allow a host cell that naturally produces the GLA, in contrast, produces DHA (such that GLA is converted to DGLA by an elongase; DGLA can then be converted to ARA by a delta-5 desaturase; ARA is then converted to EPA by a delta-17 desaturase, which can in turn be converted to DPA by an elongase; and DPA is converted to DHA by a delta-4 desaturase). In a related manner, expression of the delta-8 desaturase of the present invention allows the indirect production of ARA, EPA, DPA and / or DHA as downstream PUFAs, if subsequent desaturase and elongation reactions are catalyzed. In a preferred embodiment, where the host cell is an oil yeast, the expression cassettes encoding each of the enzymes necessary for PUFA biosynthesis will have to be inserted into the organism, since the PUFAs produced naturally in these organisms are limited to those 18:2 fatty acids (ie LA) and less frequently 18:3 fatty acids (ie ALA). Alternatively, substrate feeding may be required. Vectors or DNA cassettes useful for transforming appropriate microbial host cells are well known in the art. The specific choice of sequences present in the construct depends on the desired expression products (above), the nature of the host cell, and the proposed means for separating transformed cells. versus non-transformed cells. Typically, however, the vector or cassette contains sequences that direct transcription and translation of the relevant gene(s), a selectable marker, and sequences that allow for autonomous replication or chromosomal integration. Suitable vectors comprise a 5' region of the gene that controls transcriptional initiation and a 3' region of the DNA fragment that controls transcriptional termination. It is preferred that both control regions are derived from the genes of the transformed host cell, although it should be understood that such control regions need not be derived from genes native to the specific species chosen as a production host. Initiation control regions or promoters that are useful for directing expression of desaturase and / or elongase ORFs in the desired microbial host cell are numerous and familiar to those skilled in the art. Virtually any promoter capable of directing the expression of these genes in the selected host cell is suitable for the present invention. Expression in a microbial host cell can be carried out in a transient or stable manner. Transient expression can be accomplished by inducing an activity of a regulatable promoter operably linked to the gene of interest. Stable expression can be achieved by using a constitutive promoter operably linked to the gene of interest. As an example, when the host cell is a yeast, transcriptional and translational regions functional in yeast cells are provided, particularly from the host species. Transcriptional initiation regulatory regions can be obtained, for example, from: (1) genes in the glycolytic pathway, such as alcohol dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase (WO 2005 / 003310), phosphoglycerate mutase (WO 2005 / 003310), fructose bisphosphate aldolase (WO 2005 / 049805), phosphoglucose isomerase, phosphoglycerate kinase, glycerol-3-phosphate O-acyltransferase (see US Patent Application 60 / 610060), etc.; or (2) regulatory genes such as acid phosphatase, lactase, metallothionein, glucoamylase, EF1-a protein translation elongation factor (TEF) (US 6,265,185), S7 ribosomal protein (US 6,265,185), etc.; Any one of a number of regulatory sequences can be used, depending on whether constitutive or induced transcription is desired and depending on the promoter's efficiency in expressing the ORF of interest, ease of construction, and the like. The nucleotide sequences surrounding the 'ATG' translation initiation codon have been found to affect expression in yeast cells. If the desired polypeptide is poorly expressed in yeast, the nucleotide sequences of exogenous genes can be modified to include an efficient yeast translation initiation sequence to obtain the most favorable expression of the gene. For expression in yeast, this can be done by site-directed mutagenesis of an inefficiently expressed gene by fusing it in-frame to an endogenous yeast gene, preferably a highly expressed gene. Alternatively, as demonstrated in the present invention in Yarrowia lipolytica, it is possible to determine the consensus translation initiation sequence in the host and engineer this sequence into heterologous genes for their most favorable expression in the host of interest. The termination region can be derived from the 3' region of the gene from which the initiation region was obtained or from a different gene. A large number of termination regions are known and function satisfactorily in a variety of hosts (when used in the same genera and species and in different genera and species from which they were derived). The termination region is generally selected more as a matter of convenience than because of any particular property. Preferably, the termination region is derived from a gene for yeast, particularly Saccharomyces, Schizosaccharomyces, Candida, Yarrowia or Kluyveromyces. The 3' regions of mammalian genes encoding γ-interferon and α-2-interferon are also known to function in yeast. Termination control regions can also be derived from various genes native to preferred hosts. Optionally, a termination site may be unnecessary; however, it is preferable if it is included. As one skilled in the art is aware, the mere introduction of a gene into a cloning vector does not guarantee that it will be successfully expressed at the required level. In response to the need for a high expression rate, many specialized expression vectors have been created by manipulating a number of different genetic elements that control aspects of transcription, translation, protein stability, oxygen limitation and secretion of the host cell. More specifically, some of the molecular features that have been manipulated to control gene expression include: (1) the nature of the relevant promoter and transcriptional terminator sequences; (2) the number of copies of the cloned gene and whether the gene is plasmid-borne or integrated into the host cell genome; (3) the final cellular position of the synthesized exogenous protein; (4) the efficiency of translation in the host organism; (5) the intrinsic stability of the cloned gene protein within the host cell; and (6) codon usage within the cloned gene such that its frequency approximates the host cell's preferred codon usage frequency. Each of these types of modifications are encompassed in the present invention as the means to further optimize the expression of PUFA biosynthetic pathway enzymes. Microbial Host Transformation Once DNA encoding a desaturase or elongase polypeptide is suitable for expression in an oleaginous yeast has been obtained, it is placed on a plasmid vector capable of autonomous replication in a host cell; or is integrated directly into the host cell genome. Integration of expression cassettes can occur randomly within the host genome or can be targeted through the use of constructs that contain regions of homology with the host genome sufficient to target recombination within the host locus. Where constructs are targeted to an endogenous locus, all or some of the transcriptional and translational regulatory regions may be provided by the endogenous locus. In the present invention, the preferred method of gene expression in Yarrowia lipolytica is through integration of linear DNA into the host genome; and integration at multiple positions within the genome can be particularly useful when high level expression of genes is desired. For this purpose, it is desirable to identify a sequence within the genome that is present in multiple copies. Schmid-Berger et al., (J. Bact. 176(9):2477-2482 (1994 )) discovered the first Ylt1 retrotransposon-like element in Yarrowia lipolytica. This retrotransposson is characterized by the presence of long terminal repeats (LTRs; each about 700 bp in length) in regions called zeta. Ylt1 and solo zeta elements were present in a dispersed manner within the genome in at least 35 copies / genome and 50-60 copies / genome, respectively; both elements were determined to function as sites of homologous recombination. Furthermore, work by Juretzek et al., (Yeast 18:97-113 (2001)) demonstrated that gene expression could be dramatically increased by targeting plasmids in the repetitive regions of the yeast genome (which use linear DNA with zeta LTR regions at both ends), compared to expression obtained using the low copy plasmid transformants. Thus, zeta-directed integration may be ideal as a means to ensure multiple integration of plasmid DNA into Y. lipolytica, thereby allowing high-level gene expression. Unfortunately, however, not all Y. lipolytica strains have zeta regions (eg, the strain identified* as ATCC 20362). When the lineage lacks such regions, it is also possible to integrate plasmid DNA comprising expression cassettes at alternate loci to achieve the desired copy number for the expression cassette. For example, preferred alternate loci include: the Ura3 locus (GenBank Accession No. AJ306421), the Leu2 gene locus (GenBank Accession No. AF260230), the Lys5 gene (GenBank Accession No. M34929), the Aco2 gene locus (GenBank Accession No. AJ001300), the Pox3 gene locus (Pox3: GenBank Accession No. XP_503244; or Aco3: GenBank Accession No. AJ001301), the delta gene locus 12 desaturase (SEQ ID NO:23), the Lip1 gene locus (GenBank Accession No. Z50020) and / or the Lip2 gene locus (GenBank Accession No. AJ012632). Advantageously, the Ura3 gene can be used repeatedly in combination with the 5-fluoroorotic acid selection (5-fluorouracil-6-carboxylic acid monohydrate; "5 FOA") (below), to easily allow the genetic modifications to be integrated into the Yarrowia genome in an easy way. Where two or more genes are expressed from separate replication vectors, it is desirable that each vector have different means of selection and should lack homology to the other constructs to maintain stable expression and to prevent reassortment of elements between constructs. The judicious choice of regulatory regions, means of selection and method of propagation of the inserted construct can be determined experimentally so that all inserted genes are expressed at the levels necessary to provide synthesis of the desired products. Constructs comprising the gene of interest can be inserted into a host cell by any standard technique. These techniques include transformation (eg, lithium acetate transformation [Methods in Enzymology, 194:186-187 (1991)]), protoplast fusion, holistic (holistic) impact, electroporation, microinjection, or any other method that introduces the gene of interest in the host cell. More specific teachings applicable to oil yeasts (i.e., Yarrowia lipolytica) include US 4,880,741 and US 5,071,764 and Chen, D.C. et al., (Appl Microbiol Biotechnol. 48(2): 232-235 (1997)). For convenience, a host cell that is manipulated by any manufacturing method to extract a DNA sequence (eg, an expression cassette) will be referred to as "transformed" or "recombinant" in the present invention. The transformed host will have at least one copy of the expression construct and may have two or more depending on whether the gene is integrated into the genome, amplified, or present on an extrachromosomal element that has multiple copy numbers. The transformed host cell can be identified by various selection techniques as described in WO 2004 / 101757. Preferred selection methods for use in the present invention are resistance to kanamycin, hygromycin, and aminoglycoside G418, as well as the ability to grow on media lacking uracil, leucine, lysine, tryptophan, or histidine. In alternative embodiments, 5-FOA is used for selection of yeast Ura mutants. The compound is toxic to yeast cells that have a functioning URA3 gene encoding orotidine 5'-monophosphate decarboxylase (OMP decarboxylase); in this way, on the basis in this toxicity, 5-FOA is especially useful for the selection and identification of Ura- mutant yeast strains (Bartel, P.L. and Fields, S., Yeast 2-Hybrid System, University of Oxford: New York, v. 7, pages 109-147, 1997). More specifically, it is possible to first knockout the native Ura3 gene to produce a strain that has an Ura- phenotype, where selection occurs on the basis of resistance to 5-FOA. Then, a set of multiple chimeric genes and a novel Ura3 gene can be integrated into a different locus of the Yarrowia genome to thereby produce a new strain that has an Ura+ phenotype. Subsequent integration would produce a new Ura3- lineage (identified again using 5-FOA selection), when the inserted Ura3 gene is knocked out. Thus, the Ura3 gene (in combination with 5-FOA selection) can be used as a selection marker in multiple rounds of transformation. After transformation, appropriate substrates for the recombinantly expressed desaturases and / or elongases (and optionally other PUFA enzymes that are expressed within the host cell) can be produced by the host naturally or transgenically, or can be supplied exogenously. Metabolic Engineering of Omega-3 and / or Omega-6 Fatty Acid Biosynthesis in Microbes Methods for manipulating biochemical pathways are well known to those skilled in the art; and it is expected that numerous manipulations are possible to maximize the biosynthesis of omega-3 and / or omega-6 fatty acids in oleaginous yeasts and particularly, in Yarrowia lipolytica. This may require metabolic engineering directly within the PUFA biosynthetic pathway or further manipulation of pathways that supply carbon to the PUFA biosynthetic pathway. In the case of manipulations within the PUFA biosynthetic pathway, it may be desirable to increase LA production to allow for increased production of omega-3 and / or omega-6 fatty acids. Introduction and / or amplification of genes encoding delta-9 and / or delta-12 desaturases can accomplish this. In order to maximize the production of omega-6 unsaturated fatty acids, the person skilled in the art is well aware that production is favored in a host microorganism that is substantially free of ALA. Thus, preferably, the host is selected or obtained by removing or inhibiting delta-15 or omega-3-type desaturase activity that allows the conversion of LA to ALA. Endogenous desaturase activity can be reduced or eliminated by, for example: (1) employing a cassette for the transcription of antisense sequences to the transcription product of delta-15 desaturase; (2) disruption of the delta-15 desaturase gene by inserting, replacing and / or deleting the whole gene or a part of the target gene; or (3) use of a host cell that naturally has (or has been mutated to have) low or no delta-15 desaturase activity. Inhibition of unwanted desaturase pathways can also be accomplished through the use of specific desaturase inhibitors such as those described in US 4,778,630. Alternatively, maximizing omega-3 fatty acid production (and minimizing omega-6 fatty acid synthesis) may be desirable. Thus, it is possible to use a host microorganism in which a delta-12 desaturase activity that allows the conversion of oleic acid to LA is removed or inhibited, using any of the means described above (see also, for example, WO 2004 / 104167 , incorporated herein entirely by reference). Subsequently, the appropriate expression cassettes would be inserted into the host, along with the appropriate substrates (e.g., ALA) for the conversion of the ALA derivatives. ALA omega-3 fatty acid (eg, STA, ETrA, ETA, EPA, DPA, DHA). In addition to the immediate PUFA biosynthetic pathway, it is expected that manipulation of several other enzymatic pathways that lead to precursor fatty acid biosynthesis may contribute to the total net biosynthesis of specific PUFAs. Identifying and manipulating these related pathways will be useful in the future. Techniques to Stimulate Desirable Biosynthetic Pathways Additional copies of the desaturase and elongase genes can be inserted into the host to increase the output of omega-3 and / or omega-6 fatty acid biosynthetic pathways. Expression of desaturase or elongase genes can also be increased at the transcriptional level by using a stronger promoter (regulated or constitutive) to cause increased expression, by removing / deleting destabilizing sequences from the mRNA or encoded protein, or by adding stabilizing sequences to the mRNA (US 4,910,141). However, another approach to increase the expression of desaturase or elongase genes, as demonstrated in the present invention, is to increase the translational efficiency of the encoded mRNAs by replacing the codons in the native gene with those of more favorable expression of the gene in the host microorganism selected. Techniques to Inhibit Unwanted Biosynthetic Pathways On the other hand, the biochemical pathways that compete with the omega-3 and / or omega-6 fatty acid biosynthetic pathways for energy or carbon or the enzymes of the native PUFA biosynthetic pathway that interfere with the production of a specific PUFA end product, they can be eliminated by disrupting the gene or by inhibiting by other means (eg, antisense mRNA). For gene disruption, an exogenous fragment of DNA (typically a selectable marker gene) is inserted into the structural gene to be disrupted in order to stop gene disruption. its coding sequence and thereby functionally inactivate the gene. Transformation of the disruption cassette into the host cell results in replacement of the functional native gene by homologous recombination with the undisrupted gene (see, for example: Hamilton et al., J. Bacterial. 171:4617-4622 (1989); Balbas et al. ., Gene 136:211-213 (1993); Gueldener et al., Nucleic Acids Res. 24:2519-2524 (1996); and Smith et al., Methods Mol. Cell. Biol. 5:270-277 (1996) ). Antisense technology is another method of inhibiting genes when the target gene sequence is known. To accomplish this, a nucleic acid segment of the desired gene is cloned and operably linked to a promoter such that the antisense RNA strand is transcribed. This construct is then inserted into the host cell and the antisense RNA strand is produced. Antisense RNA inhibits gene expression by preventing the accumulation of mRNA encoding the protein of interest. Those skilled in the art will know that special considerations are associated with the use of antisense technologies in order to reduce the expression of specific genes. For example, the appropriate level of expression of antisense genes may require the use of different chimeric genes that utilize different regulatory elements known to those skilled in the art. While targeted gene disruption and antisense technology offer effective means of inhibiting genes where the sequence is known, other less specific methodologies have been developed which are not sequence based (e.g. mutagenesis via UV / chemical radiation or the use of transposable elements / transposons, see WO 2004 / 101757). Within the context of the present invention, modulation of the expression of the fatty acid biosynthetic pathway by any of the methods described above may be useful. For example, the present invention provides methods whereby genes encoding key enzymes in Biosynthetic pathways are inserted into oleaginous yeasts for the production of omega-3 and / or omega-6 fatty acids. Expression of these genes in oil yeasts that lack naturally occurring omega-3 and / or omega-6 fatty acid biosynthetic pathways and do not coordinate the expression of these genes will be particularly useful to maximize production of preferred PUFA products using various means to the metabolic engineering of the host organism. Preferred Microbial Hosts for Producing Recombinant Omega-3 and / or Omega-6 Fatty Acids Microbial host cells for producing omega fatty acids can include microbial hosts that grow on a variety of starting materials, including simple or complex carbohydrates, organic acids and alcohols, and / or hydrocarbons over a wide range of temperatures and values. of pH. The preferred microbial hosts, however, are oil yeasts. These organisms are naturally capable of synthesizing and accumulating oil, with the oil comprising greater than about 25% of the cell dry weight, more preferably greater than about 30% of the cell dry weight, and most preferably still, more than about 40% of the cellular dry weight. Genera typically identified as oil yeast include, but are not limited to, Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon, and Lipomyces. More specifically, illustrative oil-synthesizing yeasts include: Rhodosporidium toruloides, Lipomyces starkeyii, L. lipoferus, Candida revkaufi, C. pulcherrima, C. tropicalis, C. utilis, Trichosporon pullans, T. cutaneum, Rhodotorula glutinus, R. graminis, and Yarrowia lipolytica (formerly classified as Candida lipolytica). The preferred oil yeast is Yarrowia lipolytica- and in one In a further embodiment, strains of Y. lipolytica designated as ATCC 20362, ATCC 8862, ATCC 18944, ATCC 76982 and / or LGAM S(7)1 are preferred (Papanikolaou S. and Aggelis G., Bioresour. Technol. 82(1) :43-9 (2002)). Historically, various strains of Y. lipolytica have been used for the manufacture and production of: isocitrate lyase (DD259637); lipases (SU1454852, WO 2001 / 083773, DD279267); polyhydroxyalkanoates (WO 2001 / 088144); citric acid (RU2096461, RU2090611, DD285372, DD285370, DD275480, DD227448, PL160027); erythritol (EP 770683); 2-oxoglutaric acid (DD267999); y-decalactone (US 6,451,565, FR 2734843); y-dodecalatone (EP 578388); and pyruvic acid (JP 09252790). Microbial Fermentation Processes for PUFA Production The transformed microbial host cell is grown under conditions that optimize desaturase and elongase activities and produce the best and most economical yield of preferred PUFAs. In general, media conditions that can be optimized include type and amount of carbon source, type and amount of nitrogen source, carbon to nitrogen ratio, oxygen level, growth temperature , the pH, the length of the biomass production phase, the length of the oil accumulation phase and the cell culture time. Microorganisms of interest, such as oil yeast, grow in complex media (e.g., yeast dextrose peptone extract (YPD) broth) or defined minimal media that lack a necessary component for growth and thereby force , selection of the desired expression cassettes (eg, Yeast Nitrogen Base (DIFCO Laboratories, Detroit, MI)). Fermentation media in the present invention must contain an appropriate source of carbon. Appropriate carbon sources may include, but are not limited to: monosaccharides (e.g., glucose, fructose), disaccharides (e.g. lactose, sucrose), oligosaccharides, polysaccharides (e.g. starch, cellulose or mixtures thereof), sugar alcohols (e.g. glycerol) or mixtures of renewable starting materials (e.g. whey permeate cheese curds, corn steep liquor, beet sugar molasses, barley malt). Additionally, carbon sources can include alkanes, fatty acids, fatty acid esters, monoglycerides, diglycerides, triglycerides, phospholipids and various commercial sources of fatty acids including vegetable oils (eg soybean oil) and animal fats. Additionally, the carbon source can include those sources of a carbon (eg, carbon dioxide, methanol, formaldehyde, formate, and carbon-containing amines) for which metabolic conversion to key biochemical intermediates has been demonstrated. Thus, it is contemplated that the carbon source utilized in the present invention may encompass a wide variety of carbon-containing sources and will be limited only by the choice of host organism. While all of the aforementioned carbon sources and mixtures thereof are suitable in the present invention, the preferred carbon sources are sugars and / or fatty acids. Glucose and / or fatty acids containing between 10 and 22 carbons are preferred. Nitrogen can be obtained from an inorganic source (e.g. (NH 4 ) 2SO4) or organic (e.g. urea or glutamate). In addition to the appropriate carbon and nitrogen sources, the fermentation media must also contain appropriate minerals, salts, cofactors, buffers, vitamins and other components known to those skilled in the art suitable for the growth of microorganisms and the promotion of the necessary enzymatic pathways. for the production of PUFA. Particular attention is paid to several metal ions (e.g., Mn +2 , co +2 , Zn +2 , mg +2 ) that promote the synthesis of lipids and PUFAs (Nakahara, T. et al., Ind. Appl. Single Cell Oils, D. J. Kyle and R. Colin, eds. pages 61 to 97 (1992)). Preferred growth media in the present invention are common commercially prepared media such as Yeast Nitrogen Base (DIFCO Laboratories, Detroit, MI). Other defined or synthetic growth media may also be used and the appropriate medium for the growth of specific microorganisms will be known to the person skilled in the art of microbiology or fermentation science. A suitable pH range for fermentation is typically between about pH 4.0 to pH 8.0, with pH 5.5 to pH 7.0 being preferred as the range for initial growth conditions. Fermentation can be conducted under aerobic or anaerobic conditions, with microaerobic conditions being preferred. Typically, the accumulation of high levels of PUFAs in oil yeast cells requires a two-stage process, as the metabolic state must be "balanced" between growth and fat synthesis / storage. Thus, most preferably, a two-stage fermentation process is required for the production of PUFAs in oleaginous yeast. This approach is described in WO 2004 / 101757, as well as various appropriate fermentation process designs (ie batch, fed-batch and continuous) and considerations during growth are described. Purification of Microbial PUFAs PUFAs can be found in the host microorganism as free fatty acids or in esterified forms such as acylglycerols, phospholipids, sulfolipids or glycolipids and can be extracted from the host cell by a variety of means well known in the art. A review of extraction techniques, quality analysis, and acceptability standards for yeast lipids is the review by Z. Jacobs (Critical Reviews in Biotechnology 12(5 / 6):463-491 (1992)). A brief review of downstream processing is also available from A. Singh and O. Ward (Adv. Appl. Microbiol. 45:271-312 (1997)). In general, means for purifying PUFAs can include extraction with organic solvents, sonication, supercritical fluid extraction (e.g., using carbon dioxide), saponification, and physical means such as presses, or combinations thereof. . See the teachings of WO 2004 / 101757 for further details. Description of Preferred Achievements The final objective of the work described in the present invention was the identification of an appropriate delta-8 desaturase to allow the expression of the delta-9 elongase / delta-8 desaturase pathway in plants and oleaginous yeasts. Thus, early work of the present invention attempted codon optimization of Euglena gracilis delta-8 desaturase (GenBank Accession No. AAD45877; WO 00 / 34439) for expression in Yarrowia lipolytica. Despite a different codon-optimized synthesis of three genes (i.e., "D8S-1", "D8S-2", and "D8S-3"), none of the genes was able to desaturate EDA into DGLA (Example 1). Based on these results, it was hypothesized that the previously published delta-8 desaturase sequences were incorrect. Isolation of direct Euglena gracilis delta-8 desaturase, after mRNA isolation, cDNA synthesis and PCR (Examples 2 and 3) was attempted as described below. This resulted in two similar sequences, identified in the present invention as Eg5 (SEQ ID N°: 1 and 2) and Eg 12 (SEQ ID N°: 3 and 4), both of which had significant differences when compared to the delta- 8 previously published desaturases (Example 4). Eg5 and Eg12 were cloned into a Saccharomyces cerevisiae yeast expression vector (Example 5) for functional analysis via substrate feeding assays (Example 11). This demonstrated that Eg5 and Eg 12 could desaturate EDA and ETrA to produce DGLA and ETA, respectively; Eg5 had significantly greater activity than Eg 12. Based on confirmed Eg5 delta-8 desaturase activity (SEQ ID NO:1 and 2), the Eg5 sequence is codon-optimized for expression in Yarrowia lipolytica (Example 14), thereby resulting in the synthesis of a synthetic and functional codon-optimized delta-8 desaturase designated as "D8SF" (SEQ ID NOS:112 and 113). Co-expression of the codon-optimized delta-8 desaturase of the present invention together with a codon-optimized delta-9 elongase (derived from Isochrysis galbana (GenBank Accession No.: 390174)) in Y. lipolytica allowed a synthesis of 6, 4% DGLA, with no co-synthesis of GLA (Example 16). A number of expression constructs were then created to allow the synthesis of various PUFAs in soybean, using the confirmed delta-8 desaturase sequence from Eg5, the codon-optimized delta-9 elongase from Isochrysis galbana from Yarrowia lipolytica, or the elongase from Mortierella alpina, the delta-5 desaturase from Mortierella alpina, the delta-15 desaturase from Fusarium and the delta-17 desaturase from Saprolegnia diclina, and combinations thereof (Examples 17 to 22). Expression of these constructs resulted in the production of up to about 29.9% DGLA and up to about 29.4% EPA (Examples 21 and 22, respectively). Examples The present invention is further defined in the examples below, where parts and percentages are by weight and degrees are Celsius, unless otherwise indicated. It should be understood that these examples, while indicating preferred embodiments of the present invention, are provided for illustration only. From the above discussion and these examples, the person skilled in the art can verify the essential characteristics of the present invention and without deviating from the character and scope thereof, may make various changes and modifications to the present invention to adapt it to various uses and conditions. Accordingly, various modifications of the present invention in addition to those shown and described in the present invention will become apparent to those skilled in the art from the foregoing description. Such modifications are also within the scope of the appended claims. The meaning of abbreviations is as follows: "s" means second(s), "min" means minute(s), "h" means hour(s), "d" means day(s), "pl" means microliter (s), "ml" means "milliliter(s), "I" means liter(s), "pM" means micromolar, "mM" means millimolar, "M" means molar, "mmol" means "millimole(s) , "pmole” means micromole(s), "g" means gram(s), "pg" means microgram(s), "ng" means nanogram(s), "U" means unit(s), "bp" means base pair(s) and "kB" means kilobase(s). Transformation and Cultivation of Yarrowia lipolytic a Yarrowia lipolytica strains ATCC 20362, 76982 and 90812 were purchased from the American Type Culture Collection (Rockville, MD). Y. lipolytica strains were generally grown at 28°C in YPD agar (1% yeast extract, 2% bactopeptone, 2% glucose, 2% agar). Transformation of Yarrowia lipolytica was performed according to the method of Chen, D.C. et al., (Appl. Microbiol Biotechnol. 48(2):232-235 (1997 )), unless otherwise indicated. Briefly, Yarrowia was traced onto a YPD plate and grown at 30°C for about 18 hours. Several large portions of cells were scraped off the plate and resuspended in 1 ml of transformation buffer containing: 2.25 ml 50% PEG, average molecular weight 3350; 0.125 ml 2M lithium acetate, pH 6.0; 0.125 ml of 2 M DTT and 50 pg of sheared salmon sperm DNA. Next, approximately 500 ng of the linearized plasmid DNA was incubated in 100 µl of resuspended cells and kept at 39°C for 1 hour with vortex mixing at 15 minute intervals. Cells were plated onto selection media plates and kept at 30°C for 2 to 3 days. For transformant selection, minimal medium ("MM") was generally used; the composition of MM is as follows: 0.17% Yeast Nitrogen Base (DIFCO Laboratories, Detroit, MI) without ammonium sulfate or amino acids, 2% glucose, 0.1% proline, pH 6.1. Uracil supplements were added as appropriate to a final concentration of 0.01% (thus producing "MMU" selection media prepared with 20 g / l agar). Alternatively, transformants were selected in 5-fluoroorotic acid ("FOA"; also 5-fluorouracil-6-carboxylic acid monohydrate), comprising: 0.17% nitrogen-based yeast (DIFCO Laboratories, Detroit, MI) without ammonium sulfate or amino acids, 2% glucose, 0.1% proline, 75 mg / l uracil, 75 mg / l uridine, 900 mg / l FOA (Zymo Research Corp., Orange, CA) and 20 g / l of agar. Fatty Acid Analysis of Yarrowia lipolytica For fatty acid analysis, cells were collected by centrifugation and lipids were extracted as described in Bligh, E.G. & Dyer, W.J. (Can. J. Biochem. Physiol. 37:911-917 (1959)). Methyl fatty acid esters were prepared by transesterification of the lipid extract with sodium methoxide (Roughan, G. and Nishida I. Arch Biochem Biophys. 276(1):38-46 (1990)) and subsequently analyzed with a GC from Hewlett-Packard 6890 fitted with a 30 m x 0.25 mm (i.d.) HP-INNOWAX column (Hewlett-Packard). The oven temperature was 170°C (holding for 25 minutes) to 185°C at 3.5°C / min. For direct base transesterification, the Yarrowia culture (3 ml) was harvested, washed once in distilled water and dried under vacuum in a Speed-VAC for 5 to 10 minutes. Sodium methoxide (100 µl of 1%) was added to the sample and then the sample was shaken and rocked for 20 minutes. After adding three drops of 1 M NaCl and 400 µl of hexane, the sample was shaken and rotated. The top layer was removed and analyzed by GC as described above. Example 1 Synthesis and Expression of a CODON-Optimized Delta-8 Desaturase Gene in YARROWS LIPOLYTICA In order to express the Euglena gracilis delta-8 desaturase gene (SEQ ID NO.:5 and 6, GenBank Accession No. AAD45877) in Yarrowia lipolytica, the codon usage of the delta-8 desaturase gene was optimized for expression in Y. lipolytica. A codon-optimized delta-8 desaturase gene (designated "D8S-1", SEQ ID N°:48) was designed, based on the published sequence of Euglena gracilis (SEQ ID N°:5), according to the standard of Yarrowia codon usage (WO 2004 / 101753), the consensus sequence around the 'ATG' translation initiation codon, and general RNA stability rules (Guhaniyogi, G. and J. Brewer, Gene 265(1 -2):11-23 (2001)). In addition to modification of the translation initiation site, 200 bp of the 1260 bp coding region were modified (15.9%). None of the modifications in the codon-optimized gene changed the amino acid sequence of the encoded protein (SEQ ID NO:6) except the second amino acid from 'K* to 'E' to add the Ncol site around the translation initiation codon. In Vitro Synthesis of a Codon-Optimized Delta-8 Desaturase Gene for Yarrowia The codon-optimized delta-8 desaturase gene was synthesized as follows. First, thirteen pairs of oligonucleotides were designed to extend the entire length of the coding region. codon-optimized from the E. gracilis delta-8 desaturase gene (e.g. D8-1A, D8-1B, D8-2A, D8-2B, D8-3A, D8-3B, D8-4A, D8-4B, D8-5A, D8-5B, D8-6A, D8-6B, D8-7A, D8-7B, D8-8A, D8-8B, D8-9A, D8-9B, D8-10A, D8-10B, D8- 11A, D8-11B, D8-12A, D8-12B, D8-13A and D8-13B, corresponding to SEQ ID NO: 49-74). Each pair of sense (A) and antisense (B) oligonucleotides was complementary except for a 4 bp overhang at each 5' end. Additionally, primers D8-1A, D8-3B, D8-7A, D8-9B and D8-13B (SEQ ID NO: 49, 54, 60, 65 and 74) also introduced the restriction sites Ncol, Bglll, Xho1 , Saci and Not1, respectively, for subsequent subcloning. Each oligonucleotide (100 ng) was phosphorylated at 37°C for 1 hour in a 20 µl volume containing 50 mM Tris-HCI (pH 7.5), 10 mM MgCh, 10 mM DTT, 0.5 mM of spermidine, 0.5 mM ATP and 10 U of T4 polynucleotide kinase. Each pair of sense and antisense oligonucleotides was mixed and annealed in a thermoshaker using the following settings: 95°C (2 minutes), 85°C (2 minutes), 65°C (15 minutes), 37°C (15 minutes), 24°C (15 minutes) and 4°C (15 minutes). Thus, D8-1A (SEQ ID NO: 49) was annealed to D8-1B (SEQ ID NO: 50) to produce the double-stranded product "D8-1AB". Similarly, D8-2A (SEQ ID NO:51) was annealed to D8-2B (SEQ ID NO:52) to produce the double-stranded product "D8-2AB", etc. Four separate pools of double-stranded looped oligonucleotides were then ligated as shown below: (a) Pool 1: comprised D8-1AB, D8-2AB and D8-3AB; (b) Cluster 2: comprised D8-4AB, D8-5AB and D8-6AB; (c) Cluster 3: comprised D8-7AB, D8-8AB and D8-9AB; and, (d) Cluster 4: comprised D8-10AB, D8-11AB, D8-12AB and D8-13AB. Each annealed oligonucleotide pool was mixed in a 20 µl volume with 10 U of T4 DNA ligase and the ligation reaction was incubated overnight at 16°C. The product of each ligation reaction was then used as a template to amplify the designed DNA fragment by PCR. Specifically, using the ligated "Cluster 1" mixture (i.e., D8-1AB, D8-2AB, and D8-3AB) as the template and oligonucleotides D8-1F (SEQ ID NO:75) and D8-3R (SEQ ID NO:76) as primers, the first portion of the codon-optimized delta-8 desaturase gene was amplified by PCR. PCR amplification was performed in 50 µl of total volume comprising PCR buffer containing 10 mM KCl, 10 mM (NH 4 ) 2 SO4, 20 mM Tris HCl (pH 8.75), 2 mM MgSO 4 , 0.1% Triton X-100, 100 pg / ml BSA (final concentration), 200 µM of each deoxyribonucleotide triphosphate, 10 pmols of each primer, and 1 µl of PfuTurbo DNA polymerase (Stratagene, San Diego, CA ). Amplification was performed as follows: initial denaturation at 95°C for 3 minutes, followed by 35 cycles of the following: 95°C for 1 minute, 56°C for 30 s, 72°C for 40 s. A final extension cycle of 72°C for 10 minutes was performed, followed by termination of the reaction at 4°C. The 309 bp PCR fragment was subcloned into the pGEM-T easy vector (Promega) to generate pT8(1-3). Using the ligated "Cluster 2" mixture (i.e., D8-4AB, D8-5AB, and D8-6AB) as the template and oligonucleotides D8-4F (SEQ ID NO:77) and D8-6R (SEQ ID NO: °:78) as primers, the second portion of the codon-optimized delta-8 desaturase gene was similarly amplified by PCR and cloned into the pGEM-T easy vector to generate pT8(4-6). Using the "Cluster 3" mixture (i.e., D8-7AB, D8-8AB, and D8-9AB) as the template and oligonucleotides D8-7F (SEQ ID NO: 79) and D8-9R (SEQ ID NO: :80) as primers, the third portion of the codon-optimized delta-8 desaturase gene was similarly amplified by PCR and cloned into pGEM-T-easy vector to generate pT8(7-9). 4" linker (i.e., D8-10AB, D8-11AB, D8-12AB, and D8-13AB) as the template and oligonucleotides D8-10F (SEQ ID NO: 81) and D8-13R (SEQ ID NO: :82) how primers, the fourth portion of the codon-optimized delta-8 desaturase gene was similarly amplified by PCR and cloned into the pGEM-T easy vector to generate pT8(10-13). E. coli was separately transformed with pT8(1-3), pT8(4-6), pT8(7-9) and pT8(10-13) and plasmid DNA was isolated from ampicillin resistant transformants. The plasmid DNA was purified and digested with the appropriate restriction endonucleases to release the 309 bp NcoI / Bglll fragment of pT8(1-3) (SEQ ID NO:83), the 321 bp Bglll / XhoI fragment of pT8 (4-6) (SEQ ID NO:84), the 264 bp Xhol / Sac1 fragment from pT8(7-9) (SEQ ID NO:85) and the 369 bp Sac1 / Not1 fragment from pT8( 10-13) (SEQ ID NO:86). These fragments were then combined and directionally ligated with Nco1 / Not1 digested pY54PC (SEQ ID NO:115; WO 2004 / 101757) to generate pDMW240 (Figure 5A). This resulted in a synthetic delta-8 desaturase gene ("D8S-1", SEQ ID NO:48) in pDMW240. Compared to the published delta-8 desaturase amino acid sequence (SEQ ID NO:6) from E. gracilis, the second amino acid of D8S-1 was changed from 'K' to 'E' in order to add the NcoI site in around the translation initiation codon. Another version of the synthesized gene, with the exact amino acid sequence as the published E. gracilis delta-8 desaturase sequence (SEQ ID NO:6), was constructed by in vitro mutagenesis (Stratagene, San Diego, CA) using pDMW240 as a template and oligonucleotides ODMW390 (SEQ ID NO:87) and ODMW391 (SEQ ID NO:88) as primers. The resulting plasmid was designated pDMW255 (Figure 5B). The synthetic delta-8 desaturase gene in pDMW255 has been designated as "D8S-2" and the amino acid sequence is exactly the same as the sequence described in SEQ ID NO:5. Yarrowia lipolytica strain ATCC 76982 (Leu-) was transformed with pDMW240 and pDMW255, respectively, as described in General Methods. Yeast containing the pDMW240 and pDMW255 recombinant constructs (i.e., containing D8S-1 and D8S-2 respectively) grew on EDA-supplemented MM, 20:2 (11,14). Specifically, single colonies of transforming Y. lipolytica containing pDMW240 or pDMW255 were grown in 3 ml of MM at 30°C at an OD 6 oo ~ 1.0. For substrate feeding, 100 µl of the cells were then subcultured in 3 ml of MM containing 10 µg of EDA substrate for about 24 hours at 30°C. Cells were collected by centrifugation, lipids were extracted and fatty acid methyl esters were prepared by transesterification and were subsequently analyzed with a Hewlett-Packard 6890 GC. No transformants produced DGLA from EDA and thus D8S-1 and D8S-2 were non-functional and could not desaturate EDA. The D8S-1::XPR chimeric terminus and D8S-2::XPR terminus genes are shown in SEQ ID NO:89 and 90, respectively. A difference of three amino acids between the delta-8 desaturase protein sequence deposited in GenBank (Accession No. AAD45877) and in WO 00 / 34439 or Wallis et al., (Archives of Biochem. Biophys. 365:307-316 ( 1999)) (SEQ ID NO:7 in the present invention) was found. Specifically, three amino acids did not appear in GenBank Accession No. AAD45877. Using pDMW255 as the template and ODMW392 (SEQ ID NO:91) and ODMW393 (SEQ ID NO:92) as primers, 9 bp were added into the synthetic D8S-2 gene by in vitro mutagenesis (Stratagene, San Diego, CA) , thereby producing a protein that was identical to the sequence described in WO 00 / 34439 and Wallis et al., (above) (SEQ ID NO:7). The resulting plasmid was called pDMW261 (Figure 5C). The synthetic delta-8 desaturase gene in pDMW261 was designated as "D8S-3" (SEQ ID NO:93). Following the transformation of the pDMW261 construct into Yarrowia, a similar feeding experiment using EDA was conducted, as described above. No desaturation of EDA in DGLA was observed with D8S-3. Example 2 Euglena gracilis Growth Conditions, Lipid Profile and mRNA Isolation Euglena gracilis was obtained from the laboratory of Dr. Richard Triemer at Michigan State University (East Lansing, MI). From 10 ml of the actively growing culture, a 1 ml aliquot was transferred into 250 ml of Euglena gracilis medium (Eg) in a 500 ml glass vial. Eg medium was made by combining: 1 g sodium acetate, 1 g beef extract (U126-01, Difco Laboratories, Detroit, MI), 2 g Bacto® Tryptone (0123-17-3, Difco Laboratories ) and 2 g of Bacto® Yeast Extract (0127-17-9, Difco Laboratories) in 970 ml of water. After filter sterilization, 30 ml of Soil-Water Supernatant (Catalog 15-3790, Carolina Biological Supply Company, Burlington, NC) was aseptically added to produce the final Eg medium. Cultures of Euglena gracilis were grown at 23°C with a 16 hour light cycle and an 8 hour dark cycle for 2 weeks without any agitation. After 2 weeks, 10 ml of the culture was removed for lipid analysis and centrifuged at 1800 x g for 5 minutes. The pellet was washed once with water and centrifuged again. The resulting pellet was dried for 5 minutes under vacuum, resuspended in 100 µl of trimethylsulfonium hydroxide (TMSH) and incubated at room temperature for 15 minutes with agitation. After that, 0.5 ml of hexane was added and the flasks were incubated for 15 minutes at room temperature with shaking. The methyl fatty acid esters (5 µl injected from the hexane layer) were separated and quantified using a Hewlett-Packard 6890 Gas Chromatograph fitted with an Omegawax 320 fused silica capillary column (Catalog 24152, Supelco Inc.). The oven temperature was programmed to be maintained at 220°C for 2.7 minutes, increasing to 240°C at 20°C / minute and maintaining then for an additional 2.3 minutes. Carrier gas was provided by a Whatman hydrogen generator. Retention times were compared to those for methyl esters of commercially available standards (Catalog U-99-A, Nu-Chek Prep, Inc.) and the resulting chromatogram is shown in Figure 1. The remaining 2-week culture (240 ml) was pelleted by centrifugation at 1800 x g for 10 minutes, washed once with water and centrifuged again. Total RNA was extracted from the resulting pellet using STAT-60® RNA reagent (TEL-TEST, Inc., Friendswood, TX) following the protocol provided by the manufacturer (using 5 ml of the RNA reagent dissolved in 0.5 ml of water) . In this way, 1 mg of total RNA (2 mg / ml) was obtained from the pellet. mRNA was isolated from 1 mg of total RNA using the mRNA Purification Kit (Amersham Biosciences, Piscataway, NJ) following the protocol provided by the manufacturer. In this way, 85 µg of mRNA was obtained. Example 3 cDNA and PCR synthesis of Delta-8 desaturase from Euglena gracilis cDNA was synthesized from 765 ng of mRNA (Example 2) using the Superscript® Choice System for cDNA synthesis (Invitrogen® Life Technologies, Carlsbad, CA) with the supplied oligo(dT) primer according to the protocol of the manufacturer. The synthesized cDNA was dissolved in 20 µl of water. Euglena gracilis delta-8 desaturase was amplified from cDNA with oligonucleotide primers Eg5-1 (SEQ ID NO: 8) and Eg3-3 (SEQ ID NO:9) using the conditions described below. cDNA (1 µl) from the reaction described above was combined with 50 pmol Eg5-1, 50 pmol Eg5-1, 1 µl PCR nucleotide mix (10 mM, Promega, Madison, WI), 5 µl 10X PCR buffer (Invitrogen), 1.5 µl MgCl 2 (50 mM, Invitrogen), 0.5 µl of Taq polymerase (Invitrogen) and water to complete 50 µl. The reaction conditions were 94°C for 3 minutes, followed by 35 cycles of 94°C for 45 s, 55°C for 45 s, and 72°C for 1 minute. PCR was terminated at 72°C for 7 minutes and then held at 4°C. The PCR reaction was analyzed by electrophoresis with agarose gel in 5 pl and a DNA band with molecular weight around 1.3 kB was observed. The remaining 45 µl of product was separated by agarose gel electrophoresis and the DNA strand was purified using the Zymoclean® DNA Recovery Kit (Zymo Research, Orange, CA) according to the manufacturer's protocol. . The resulting DNA was cloned into pGEM®-T Easy Vector (Promega), according to the manufacturer's protocol. Multiple clones were sequenced using T7 (SEQ ID NO: 10), M13-28Rev (SEQ ID NO: 11), Eg3-2 (SEQ ID NO: 12) and Eg5-2 (SEQ ID NO: 12). : 13). Therefore, two classes of DNA sequences were obtained, Eg5 (SEQ ID N°: 1) and Eg 12 (SEQ ID N°: 3), which differed only by a few bp. Translation of Eg5 and Eg 12 gave rise to protein sequences that differed by only one amino acid, SEQ ID NO: 2 and 4, respectively. Therefore, the DNA and protein sequences for Eg5 are shown in SEQ ID NO: 1 and SEQ ID NO: 2, respectively; DNA and protein sequences for Eg12 are shown in SEQ ID NO: 3 and SEQ ID NO: 4, respectively. Example 4 Comparison of Polypeptide Sequences Shown in SEQ IP No.: 2 and 4 for Published Sequences of Delta-8 Desaturase from Euglena gracilis An alignment of the protein sequences shown in SEQ ID NO: 2 and SEQ ID NO: 4 with the protein sequence from GenBank Accession No. AAD45877 (gi: 5639724) and the published protein sequences of Wallis et al., (Archives of Biochem. Biophys., 365: 307-316 (1999); document WO 00 / 34439) is shown in Figure 2. Amino acids conserved between all four sequences are indicated with an asterisk (*). Dashes are used by the program to maximize sequence alignment. The cytochrome b domain 5 putative is underlined. A putative His box is shown in bold. It is evident that there are significant differences between the sequences of the present invention and those previously described. Specifically, the N-terminus has multiple amino acid changes. Compared to SEQ ID NO: 2, the amino acid sequences have an extra serine between L9 and P10 and amino acids from position T12 to T16 are completely different ('TIDGT to 'QLMEQ'). These changes result from multiple insertions into the DNA sequence of the published sequence and this causes three structural changes in this region. These changes are only 10 amino acids away from the cytochrome b domain 5 putative ('HPGG'). In addition, there are seven other single amino acid changes (S50 to F, S67 to F, W177 to C, L203 to P, S244 to C, T278 to A, S323 to P) with the change in W177 being only four amino acids away away from the second putative His box ('HNAHH'). Surprisingly, the protein sequence published from GenBank is missing three amino acids (S20, A21, W22) compared to that for either SEQ ID NO: 2, SEQ ID NO: 4 or WO 00 / 34439. The DNA sequence shown in WO 00 / 34439 encodes a protein identical to AAD45877 (ie lacking these three amino acids) and not the protein sequence described in WO 00 / 34439. Interestingly, the protein sequence shown in SEQ ID NO: 4 has a single amino acid change compared to SEQ ID NO: 2 (T278 to A). In Table 4, the percentage of identities between the delta-8 protein sequence Functional desaturase from Euglena gracilis claimed in the present invention (SEQ ID NO: 2) and the published sequences (SEQ ID NO: 6 and 7) are shown. Table 4 PERCENTAGE OF IDENTITY OF DELTA-8 AMINO ACID SEQUENCES Euglena gracilis Desaturase and Euglena Homologous Polypeptides gracilis % identity with % identity with SEQ ID NO: 6 SEQ ID NO: 7 SEQ ID NO. 2 95.5 96.2 * “% Identity” is defined as the percentage of amino acids that are identical between the two proteins. Calculations of sequence alignments and percent identity were performed using the Megalign program from the Lasergene bioinformatics computing package (Dnastar Inc., Madison, WI). Multiple sequence alignment was performed using the Clustal alignment method (Higgins and Sharp, Cabios. 5: 151-153 (1989)) with default parameters (Gap penalty =10, Gap length penalty = 10). The default parameters for pairwise alignments using the Clustal method were ktuple 1, Gap Penalty = 3, Window = 5, and Saved Diagonals = 5. Example 5 Cloning of Euglena gracilis Delta-8 Desaturase into a Yeast Expression Vector The episomal plasmid yeast vector pRS425 of the type (YEp) (Christianson et al., Gene, 110: 119-122 (1992)) contains 2p sequences from endogenous Saccharomyces cerevisiae plasmid, an LEU2 selectable marker, and backbone-based sequences of a phagemid (phagemid) all-in-one, pBluescript II SK+. The constitutive strong glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter from S. cerevisiae was cloned between the SacII and SpeI sites of pRS425 in the same manner as described in Jia et al., (Physiological Genomics, 3: 83-92 ( 2000)) to produce pGPD-425. A NotI site was introduced into the BamHI site of pGPD-425, thereby producing a NotI site flanked by BamHI sites, thereby resulting in plasmid pY-75. Eg5 (SEQ ID NO: 1) and Eg12 (SEQ ID NO: 3) were released from the pGEM®-T Easy vectors described in Example 2 by digestion with NotI and cloned into the NotI site of pY-75 to produce pY89- 5 and pY89-12, respectively. Thus, delta-8 desaturases (ie, Eg5 [SEQ ID NO: 1] and Eg12 [SEQ ID NO: 3]) were cloned behind a strong constitutive promoter for expression in S. cerevisiae. A map of pY89-5 is shown in Figure 3A. Example 6 Cloning of Euglena gracilis Delta-8 Desaturase into a Soybean Expression Vector and Co-Expression with an Elongase from Mortierella ALPINA A starting plasmid pKS123 (WO 02 / 08269, the contents of which are incorporated herein by reference) contains the hygromycin B phosphotransferase (HPT) gene [ Gritz, L. and Davies, J. Gene 25: 179-188 (1983 )], flanked by the T7 promoter and transcription terminator (T7prom / hpt / T7term cassette), and a bacterial origin of replication (ori) for selection and replication in bacteria (eg E. coli). In addition, pKS123 also contains the hygromycin B phosphotransferase gene, flanked by the 35S promoter (Odell et al., Nature 313: 810-812 (1985)) and NOS 3' transcription terminator (Depicker et al., J. Mol Appt Genet 1: 561-570 (1982) (35S / hpt / NOS3' cassette) for selection in plants such as soybean. pKS123 also contains a NotI restriction site, flanked by the promoter for the α' subunit of 0- conglycinin ( Beachy et al., EMBO J. 4: 3047 A 3053 (1985 )) and the 3' transcription termination region of the phaseolin gene ( Doyle, J. J. et al., J. Biol. Chem. 261: 9228 a 9238 (1986) thereby enabling strong tissue-specific expression in soybean seeds of cloned genes at the NotI site. The pKR72 vector is a derivative of pKS123, in which the HindIII fragment that contains the P-conglycinin / A / ot / / phaseolin cassette has been inverted, and a sequence (SEQ ID NO: 14) that contains the P-conglycinin / A / ot / / phaseolin cassette restriction Sbfl, Fsel and BsiWI, was introduced between the HindIII and BamHI sites in front of the 0-conglycinin promoter. The gene for Mortierella alpina elongase was amplified from pRPB2 (WO 00 / 12720) using primers RPB2forward (SEQ ID NO: 15) and RPB2reverse (SEQ ID NO: 16), which were designed to introduce sites of NotI restriction enzyme on both ends of the elongase. The resulting PCR fragment was digested with NotI and cloned into the NotI site of pKR72 to produce pKR324. The pKS121 vector (WO 02 / 00904) contains a NotI site flanked by the Kunitz Soybean Trypsin Inhibitor (KTi) promoter (Jofuku et al., Plant Cell 1: 1079-1093 (1989)) and the termination region KTi 3', the isolation of which is described in US 6,372,965 (KTi / A / ot / / KTi3' cassette). The pKR457 vector is a derivative of pKS121, in which the upstream and downstream restriction sites of the Kti / / Vof / / Kti3' cassette have been altered through several subcloning steps. The pKR457 vector also contains the soy albumin transcription terminator downstream of the Kti terminator to extend and enhance transcription termination. In pKR457, the BamHI site upstream of the Kti promoter in pKS121 has been removed and a new sequence (SEQ ID NO: 17) has been added, containing a BsiWI, Sail, Sbfl and Hindlll site, with the BsiWI site being closer to the end 5' from the Kti promoter. In addition, the Sall site downstream of the Kti terminator in pKS121 has been removed, and a new sequence (SEQ ID NO: 18) has been added, containing an XbaI site (closer to the 3' end of the Kti terminator), a BamHI site, the soybean albumin transcription terminator sequence, a BsiWI site and another BamHI site. The albumin transcription terminator was previously amplified from soybean genomic DNA using the oSalb-12 primer (SEQ ID NO: 19; designed to introduce the BamHI, Xbal and BsiWI sites at the 3' end of the terminator), and oSalb-13 primer (SEQ ID NO: 20, designed to introduce BamHI sites at the 5' end of the terminator). After PCR, the sites at the ends were modified by subcloning through various intermediate vectors to finally produce the sequence shown in SEQ ID NO: 5. Eg5 (SEQ ID NO: 1) was released from pGEM®-T Easy by digestion with NotI and cloned into the NotI site of pKR457 to produce pKR680. Plasmid pKR680 was then digested with BsiWI, and the fragment containing Eg5 (SEQ ID NO: 1) was cloned into the BsiWI site of pKR324 (WO 2004 / 071467), to produce pKR681. Therefore, delta-8 desaturase (Eg5; SEQ ID NO: 1) could be co-expressed with Mortierella alpina elongase behind strong seed-specific promoters. A map of pKR681 is shown in Figure 3B. Example 7 Isolation of Specific Promoters for Soybean Seed Soybean annexin promoters and BD30 were isolated with the Universal GenomeWalker system (Clontech) according to its user manual (PT3042-1). To obtain soybean GenomeWalker libraries, samples of soybean genomic DNA were digested with Dral, EcoRV, Pvull and Stul separately for 2 hours. After DNA purification, the digested genomic DNAs were ligated into the GenomeWalker AP1 and AP2 adapters. Two gene-specific primers (ie, GSP1 [SEQ ID NO: 21] and GSP2 [SEQ ID NO: 22]) were designed for the soybean annexin gene based on available 5' annexin cDNA coding sequences. in an EST database (E.I. duPont de Nemours and Co., Inc., Wilmington, DE). API and GSP1 primers were used in the first round of PCR, using the conditions defined in the GenomeWalker system protocol. Cycle conditions were 94°C for 4 minutes; 94°C for 2 s and 72°C for 3 minutes, 7 cycles; 94°C for 2 s and 67°C for 3 minutes, 32 cycles; 67°C for 4 minutes. First run PCR products were diluted 50-fold. 1 pl of the diluted products was used as templates for the second PCR run with AP2 and GSP2 primers. Cycle conditions were 94°C for 4 minutes, 94°C for 2 s and 72°C for 3 minutes, 5 cycles; 94°C for 2 s and 67°C for 3 minutes, 20 cycles; 67°C for 3 minutes. A 2.1 kB genomic fragment was amplified and isolated from the GenomeWalker library digested with EcoRV. The genomic fragment was digested with BamHI and Sail and cloned in Bluescript KS vector + for sequencing. The DNA sequence of this 2012 bp soybean annexin promoter fragment is shown in SEQ ID NO: 29. Based on this sequence, two oligonucleotides with BamHI or NotI sites at the 5' ends were designed to reamplify the promoter (i.e. , SEQ ID Nos. 30 and 31). Two gene-specific primers (GSP3 [SEQ ID NO. 23] and GSP4 [SEQ ID NO. 24]) were designed to amplify the soybean BD30 promoter based on the 5' BD30 cDNA coding sequences in GenBank ( Accession No. J05560). The AP1 and GSP3 primers were used in the first round of PCR, using the same conditions defined in the GenomeWalker system protocol; however the cycle conditions used for the growth promoter soy annexin did not work well for the soy BD30 promoter. A modified contact PCR protocol was used, in which the cycle conditions were: 94°C for 4 minutes; 94°C for 2 s and 74°C for 3 minutes, 6 cycles, where the annealing temperature drops by 1°C each cycle; 94°C for 2 s and 69°C for 3 minutes, 32 cycles; 69°C for 4 minutes. First run PCR products were diluted 50-fold. 1p of the diluted products was used as templates for the second PCR run with primers AP2 and GSP4. The cycle conditions were: 94°C for 4 minutes, 94°C for 2 s and 74°C for 3 minutes, 6 cycles, where the annealing temperature drops by 1°C at each cycle; 94°C for 2 s and 69°C for 3 minutes, 20 cycles; 69°C for 3 minutes. A 1.5 kB genomic fragment was amplified and isolated from the Pvull-digested GenomeWalker library. The genomic fragment was digested with BamHI and Sail and cloned in Bluescript KS vector + for sequencing. DNA sequencing determined that this genomic fragment contained a 1408 bp soybean BD30 promoter sequence (SEQ ID NO: 25). Based on the cloned soybean BD30 promoter sequence, two oligonucleotides with BamHI or NotI sites at the 5' ends were designed to re-amplify the BD30 promoter (ie, SEQ ID NO: 32 and 33). The amplified annexin promoter and BD30 fragments (above) were digested with BamHI and NotI, purified, and cloned into the BamHI and NotI sites of plasmid pZBL115 to produce pJS88 and pJS89, respectively. Plasmid pZBL115 contains the pBR322 origin of replication, the bacterial HPT hygromycin resistance gene driven by a T7 promoter and a T7 terminator, and a promoter-HPT-Nos3' 35S gene, to serve as a resistant plant selection marker hygromycin. The M. alpina delta-6 desaturase gene was cloned into the NotI site of pJS88 and pJS89, in sense orientation, to make pJS92 and pJS93 plant expression cassettes, respectively. Based on the GenBank soy glycinin Gy1 promoter sequence sequences (Accession No. X15121), the oligonucleotides shown in SEQ ID NO: 27 and 28 were designed to amplify the soy glycinin Gy1 promoter (SEQ ID NO. °: 26), in which the primers had BamHI or Not! at the 5' ends. The amplified soybean Glykinin Gy1 promoter fragment was digested with BamHI and NotI, purified and cloned into the BamHI and NotI sites of plasmid pZBL115 (above) to produce pZBL117. Example 8 Cloning of Delta-8 Desaturase from Euglena gracilis into a Soybean Expression Vector and Co-expression with EPA Biosynthetic Genes (Delta-8 Desaturase and Delta-17 Desaturase Plasmid pKR325 was generated from pKR72 (Example 5) by HindIII digestion to remove the pcon / A / ot / / Phas3' cassette. Plasmid pKR680 (Example 5) was digested with BsiWI, and the fragment containing Eg5 (SEQ ID NO: 1) was cloned into the BsiWI site of pKR325 to produce pKR683. The pKS121 KTi / A / of / / KTi3' cassette was amplified by PCR using primers oKTi5 (SEQ ID NO: 34) and 0KTÍ6 (SEQ ID NO: 35), designed to introduce an Xbal and BsiWI site on both ends of the cassette. The resulting PCR fragment was subcloned into the Xbal site of the pUC19 cloning vector to produce plasmid pKR124, thereby adding a Pstl and Sbfl site at the 3' end of the Kti transcription terminator. The Sal I fragment from pJS93 which contained the soybean BD30 promoter (WO 01 / 68887) was combined with the Sal I fragment from pUC19 to produce pKR227, thereby adding a Pst I and Sbf I site at the 5' end of the BD30 promoter. The BD30 3' transcriptional terminator was amplified by PCR from soybean genomic DNA using the primer oSBD30-1 (SEQ ID NO: 36; designed to introduce a NotI site at the 5' end of the terminator) and primer OSBD30-2 (SEQ ID NO. 37; designed to introduce a BsiWI site at the 3' end of the terminator). The resulting PCR fragment was subcloned into the pCR-Script AMP SK(+) intermediate cloning vector (Stratagene), according to the manufacturer's protocol, to produce plasmid pKR251r. The EcoRI / / Vof / fragment of pKR251r, which contained the BD30 3' transcriptional terminator, was cloned into the EcoRI / A / ot / fragment of the intermediate cloning vector pKR227 to produce pKR256. The annexin promoter from pJS92 (Example 7) was released by digestion with BamHI and the ends were filled in. The resulting fragment was ligated into the filled-in BsiWI fragment of the pKR124 vector backbone in a direction that added a Pstl and Sbfl site at the 5' end of the annexin promoter, to produce pKR265. The annexin promoter was released from pKR265 by digestion with Sbfl and Notl, and was cloned into the Sbfl / Notl fragment of pKR256 (which contains the BD30 3' transcription terminator, an ampicillin resistance gene, and a bacterial oh region) to produce pKR268. The gene for the delta-17 desaturase of Saprolegnia diclina was released from pKS203 (Pereira et al., Biochem. J. 378; 665 to 671 (2004)) by partial digestion with NotI, and was cloned into the NotI site of pKR268, to produce pKR271. Thus, delta-17 desaturase was cloned as an expression cassette behind the annexin promoter, with the BD30 transcription terminator. Plasmid pKR271 was then digested with Pstl and the fragment containing the delta-17 desaturase Saprolegnia diclina was cloned into the Sbfl site of pKR683 to produce pKR685. Thus, delta-8 desaturase could be co-expressed with S. diclina delta-17 desaturase behind strong seed-specific promoters. A map of pKR685 is shown in Figure 4A. Example 9 Assembly of EPA Biosynthetic Pathway Genes for Expression in Somatic Soybean Embryos and Soybean Seeds (Delta-6 Desaturase, Elongase and Delta-5 Desaturase) M. alpina delta-6 desaturase (US 5,968,809), M. alpina elongase (WO 00 / 12720) and M. alpina delta-5 desaturase (US 6,075,183) were cloned into plasmid pKR274 (Figure 4B) behind strong seed-specific promoters, allowing for high expression of these genes in somatic soybean embryos and soybean seeds. All of these promoters exhibit strong tissue-specific expression in soybean seeds. Plasmid pKR274 also contains the hygromycin B phosphotransferase gene (Gritz, L. and Davies J., Gene 25: 179-188 (1983)) cloned behind the T7 RNA polymerase promoter and followed by the T7 terminator (T7prom / HPT / cassette T7term) for plasmid selection on hygromycin B in certain E. coli strains (eg, NovaBlue(DE3) (Novagen, Madison, WI), a strain that is lysogenic for lambda DE3 and carries the T7 RNA polymerase gene under control of lacUV5). In addition, plasmid pKR274 contains a bacterial origin of replication (ori) functional in E. coli from vector pSP72 (Stratagene). More specifically, the delta-6 desaturase was cloned behind the promoter for the α'-conglycinin subunit (Beachy et al., Embo J. 4: 3047-3053 (1985)) followed by the 3' transcription termination region of the phaseolin gene ( Doyle, J. J. et al. J. Biol. Chem. 261: 9228-9238 (1986 )) (Pcon / Mad6 / Phas3' cassette). The delta-5 desaturase was cloned behind the Kunitz soybean trypsin inhibitor (KTi) promoter (Jofuku et al., Plant Cell 1: 1079-1093) (1989)), followed by the KTi 3' termination region, the isolation of which is described in US 6,372,965 (KTi / Mad5 / KTi3' cassette). Elongase was cloned behind the Gy1 glycinin promoter followed by by the 3' termination region of pea legumin A2 (Gy1 / Maelo / legA2 cassette). The gene for M. alpina delta-6 desaturase was amplified by PCR from pCGR5 (US 5,968,809) using primers oCGR5-1 (SEQ ID NO: 38) and oCGR5-2 (SEQ ID NO: 38). 39), which were designed to introduce restriction enzyme NotI sites at both ends of the delta-6 desaturase and an NcoI site at the start codon of the reading frame for the enzyme. The resulting PCR fragment was subcloned into the intermediate pCR-Script AMP SK(+) cloning vector (Stratagene), according to the manufacturer's protocol, to produce plasmid pKR159. The NotI fragment of pKR159, which contained the M. alpina delta-6 desaturase gene, was cloned into the NotI site of pZBL117 (Example 7) in sense orientation to produce the pZBL119 plant expression cassette. The pKR197 vector was constructed by combining the Asci fragment of plasmid pKS102 (WO 02 / 00905), which contains the T7prom / hpt / T7term cassette and bacterial ori, with the Asci fragment of plasmid pKR72 (Example 5), which contains the pcon / cassette / Vof / / Phas. Plasmid pKR159 was digested with NotI to release the M. alpina delta-6 desaturase, which was in turn cloned into the NotI site of the soybean expression vector pKR197 to produce pKR269. The glycinin Gy1 promoter was amplified from pZBL119 using the oSGIy-1 primer (SEQ ID NO: 40; designed to introduce an Sbfl / Pstl site at the 5' end of the promoter) and the oSGIy-2 primer (SEQ ID NO: 40; °: 41; designed to introduce a NotI site at the 3' end of the promoter). The resulting PCR fragment was subcloned into the intermediate pCR-Script AMP SK(+) cloning vector (Stratagene) according to the manufacturer's protocol to produce plasmid pSGIy12. The legA2 promoter was amplified from pea genomic DNA using the primer LegPro6' (SEQ ID NO: 42; designed to introduce the Xbal and BsiWI sites at the 5' end of the promoter) and the primer LegPro3' (SEQ ID NO: 43; designed to introduce a NotI site at the 3' end of the promoter). The legA2 transcriptional terminator was amplified from pea genomic DNA using the LegTermõ' primer (SEQ ID NO: 44; designed to introduce the NotI site at the 5' end of the terminator) and the LegTerm3' primer (SEQ ID NO: 44; 45; designed to introduce BsiWI and XbaI sites at the 3' end of the terminator). The resulting PCR fragments were then combined and amplified again using LegPro5' and LegTerm3' primers, thereby forming a legA2 / / Vot / / legA23' cassette. The legA2 / / Vot / / legA23' cassette PCR fragment was subcloned into the pCR Script AMP SK(+) intermediate cloning vector (Stratagene), according to the manufacturer's protocol, to produce plasmid pKR140. Plasmid pKR142 was constructed by cloning the BsiWI fragment from pKR140 (which contains the legA2 / A / of / / legA23' cassette) into the BsiWI site of pKR124 (which contains a bacterial ori and ampicillin resistance gene). The PstIINotl fragment from plasmid pKR142 was then combined with the PstIINotl fragment from plasmid pSGIy12 (which contains the Gy1 glycinin promoter) to produce pKR263. The gene for M. alpina delta-5 desaturase was amplified from pCGR4 (US 6,075,183) using primers CGR4forward (SEQ ID NO: 46) and CGR4reverse (SEQ ID NO: 47), which were designed to introduce the NotI restriction enzyme sites at both ends of the desaturase. The resulting PCR fragment was digested with NotI and cloned into the NotI site of the pKR124 vector (Example 6) to produce pKR136. The NotI fragment containing the M. alpina elongase (Example 5) was cloned into the NotI site of vector pKR263 to produce pKR270. The Gy1 / Maelo / legA2 cassette was released from plasmid pKR270 by digestion with BsiWI and Sbfl and was cloned into the BsiWIlSbfl sites of plasmid pKR269 (which contains the delta-6 desaturase, the T7prom / hpt / T7term cassette and the bacterial ori region). This was designated as plasmid pKR272. The KTi / Mad5 / KTi3' cassette, released from pKR136 by digestion with BsiWI, was then cloned into the BsiWI site of pKR272 to produce pKR274 (Figure 4B). Example 10 Assembly of EPA Biosynthetic Pathway Genes for Expression in Somatic Soybean Embryos and Soybean Seeds (Delta-17 Desaturase and Delta-5 Desaturase) In a similar manner to that described in Example 9, the S. diclina delta-17 desaturase could be cloned into a soybean expression vector with the M. alpina delta-5 desaturase. The annexin / delta17 / BD30 cassette from pKR271 could be released by digestion with a suitable restriction enzyme such as Pstl and cloned into a soybean expression vector that already carries the M. alpina delta-5 desaturase behind a suitable promoter and a suitable selection marker, such as hygromycin. M. alpina delta-5 desaturase could be part of any suitable expression cassette described herein. For example, the NotI fragment containing the M. alpina delta-5 desaturase described above could be cloned into the NotI site of the pKR263 Gy1 / Afof / / legA2 cassette. This Gy1 / delta5 / legA2 cassette could then be cloned into a vector that contains a selectable marker suitable for soybean transformation. This vector could be co-transformed in soybean with pKR681 (Example 6) and the transforming expression genes from both selected plasmids. Thus, EPA could be produced using the delta-8 pathway independent of a delta-6 desaturase. Example 11 Functional Analysis of Delta-8 Desaturase from Euglena gracilis in Saccharomyces cerevisiae Plasmids pY89-5 (which comprise the Eg5 sequence; see Figure 3A and ATCC PTA 6048), pY89-12 (identical to pY89-5 except that the Eg12 sequence was inserted instead of Eg5) and pY-75 (Example 5, negative control cloning vector ( lacking Eg5 or Eg 12)) were transformed into Saccharomyces cerevisiae BY4741 (ATCC 201388) using standard lithium acetate transformation procedures. Transformants were selected in DOBA medium supplemented with CSM-leu (Qbiogene, Carlsbad, CA). Transformants from each platelet were inoculated into 2 ml of DOB medium supplemented with CSM-leu (Qbiogene) and cultured for 1 day at 30°C, after which 0.5 ml was transferred to the same medium supplemented with either EDA or EtrA to 1 mM. These were incubated overnight at 30°C, 250 rpm, the pellets were obtained by centrifugation and dried under vacuum. Pellets were transesterified with 50 µl TMSH and analyzed by GC as described in Example 1. Two clones for pY-75 (i.e., clones 75-1 and 75-2) and pY89-5 (i.e., clones 5-6-1 and 5-6-2) were analyzed, while two sets of clones for pY89-12 (i.e. clones 12-8-1, 12-8-2, 12-9-1 and 12- 9-2) of two independent transformations were analyzed. The lipid profile obtained by GC analysis of EDA-fed clones is shown in Table 5; and the lipid profile obtained by GC analysis of EtrA-fed clones are shown in Table 6. Table 5 Clone 16:0 16:1 18:0 18:1 20:2 20:3 (8,11,14) % 20:2 Converted 75-1 14 32 5 38 10 0 0 75-2 14 31 5 41 9 0 0 5-6-1 14 32 6 40 6 2 24 Clone 16:0 16:1 18:0 18:1 20:2 20:3 (8,11,14) % 20:2 Converted 5-6-2 14 30 6 41 7 2 19 12-8-1 14 30 6 41 9 1 7 12-8-2 14 32 5 41 8 1 8 12-9-1 14 31 5 40 9 1 8 12-9-2 14 32 5 41 8 1 7 Table 6 Clone 16:0 16:1 18:0 18:1 20:3 (11,14,17) 20:4 (8,11,14,17) % 20:3 Converted 75-1 12 25 5 33 24 0 0 75-2 12 24 5 36 22 1 5 5-6-1 13 25 6 34 15 7 32 5-6-2 13 24 6 34 17 6 ​​27 12-8-1 12 24 5 34 22 2 8 12-8- 2 12 25 5 35 20 2 9 12-9-1 12 24 5 34 22 2 9 12-9-2 12 25 6 35 20 2 9 The data in Tables 4 and 5 showed that delta-8 Euglena desaturase can desaturate EDA and EtrA. The sequence shown in SEQ ID NO: 4 has a 5 amino acid change compared to the sequence shown in SEQ ID NO: 2 and has reduced delta-8 desaturase activity. The small amount of 20:4 (8, 11, 14, 17) generated by clone 75-2 in Table 6 had a slightly different retention time than a standard for 20:4 (8, 11, 14, 17). This peak was most likely a small amount of a different fatty acid generated by yeast of the type wild in that experiment. Example 12 Cloning of Other Delta-8 Desaturases or Elongases into Soybean Expression Vectors In addition to the delta-8 desaturase from Euglena gracilis, other delta-8 desaturases can be cloned into soybean expression vectors, such as those described in Example 6 and Example 8. For example, a suitable delta-8 desaturase from an organism that no Euglena gracilis can be cloned using methods similar, without limitation, to the methods described in Example 2 and Example 3. PCR primers designed to introduce NotI sites at the 5' and 3' ends of the delta-8 desaturase can be used to amplify the gene. The resulting PCR product can then be digested with NotI and cloned into a soybean expression vector such as pKR457. Further subcloning into other vectors, as described in Example 6 or Example 8, would produce vectors suitable for expression and co-expression of delta-8 desaturase in soybean. Likewise, in addition to elongase from Mortierella alpina, other elongases can be cloned into soybean expression vectors, such as those described in Example 6 and Example 8. Specifically, elongases with specificity for linolenic acid or alpha-linolenic acid, such as such as from Isochrysis galbana (WO 2002 / 077213) can be used. For example, a suitable elongase from an organism other than Mortierella alpina can be cloned using methods similar, without limitation, to the methods described in Example 2 and Example 3. PCR primers designed to introduce NotI sites at the ends 5' and 3' elongase can be used to amplify the gene. The resulting PCR product can then be digested with NotI and cloned into soy expression vectors such as pKR72 or pKR263. More subcloning in other vectors as described in Example 6 or Example 8 would produce suitable vectors for expression and co-expression of elongase in soybean. Example 13 Transformation of Somatic Soybean Embryo Cultures Culture Conditions Soybean (cv. Jack) embryogenic suspension cultures can be maintained in 35 ml of SB196 liquid medium (below) on a gyratory shaker, 150 rpm, 26°C with cold white fluorescent light at 16:8 h day / night photoperiod at light intensity from 60 to 85 pE / m 2 / s. Cultures are subcultured every 7 days to 2 weeks by inoculating about 35 mg of tissue in 35 ml of fresh SB196 fluid (preferred subculture interval is every 7 days). Soybean embryogenic suspension cultures can be transformed with the plasmids and DNA fragments described above by the particle gun bombardment method (Klein et al., Nature, 327: 70 (1987)) using a Biolistic PDS1000 instrument. DuPont / HE (helium retrofit) for all transformations. Initiation of the Soybean Embryogenic Suspension Culture Soybean crops are started twice each month with 5 to 7 days between each start. Immature seed pods from soybean plants available 45 to 55 days after planting are harvested, removed from their husks and placed in a sterilized magenta box. The soybean seeds are sterilized by shaking them for 15 minutes in a 5% Clorox solution with 1 drop of ivory soap (i.e. 95 ml of autoclave distilled water plus 5 ml of Clorox and 1 drop of soap, mixed well ). The seeds are rinsed using two 1 liter bottles of sterile distilled water and those with less of 4 mm were placed on individual microscope slides. The small end of the seed is cut off, and the cotyledons are pressed out of the seed coat. Cotyledons are transferred to platelets containing SB1 medium (25 to 30 cotyledons per platelet). Platelets are wrapped with fiber tape and stored for 8 weeks. After that time, the secondary embryos are cut and placed in SB196 liquid medium for 7 days. DNA Preparation for Bombardment An intact plasmid or a fragment of plasmid DNA containing the genes of interest and the selectable marker gene can be used for bombardment. Fragments of plasmids such as pKR274 and pKR685 or pKR681 and / or other expression plasmids can be obtained by gel isolation of digested plasmids. In each case, 100 pg of plasmid DNA can be used in 0.5 ml of the specific enzyme mixture described below. Plasmids could be digested with Ascl (100 units) in Buffer NE 4 (20 mM Tris-acetate, 10 mM magnesium acetate, 50 mM potassium acetate, 1 mM dithiothreitol, pH 7.9) 100 pg / ml of BSA and 5 mM of beta-mercaptoethanol at 37°C for 1.5 hours. The resulting DNA fragments could be separated by electrophoresis in 1% SeaPlaque GTG agarose (BioWhitaker Molecular Applications) and the DNA fragments containing EPA biosynthetic genes could be cut from the agarose gel. The DNA could be purified from agarose using a GELase digestion enzyme, according to the manufacturer's protocol. Alternatively, whole plasmids or a combination of whole plasmid and fragment could be used. A 50 µl aliquot of sterile distilled water containing 3 mg of gold particles could be added to 5 µl of a 1 pg / pl DNA solution (either intact plasmid or DNA fragment prepared as described above). above), 50 µl of 2.5 M CaClfe and 20 µl of 0.1 M spermidine. The mixture is vortexed for 3 minutes on level 3 and spun for 10 seconds in a bench top microcentrifuge. After washing with 400 µl of 100% ethanol, the pellet is suspended by sonication in 40 µl of 100% ethanol. 5 µl of DNA suspension is applied to each flywheel of the Biolistic PDS1000 / HE instrument disk. Each 5 pl aliquot contained about 0.375 mg of gold particles per sputter (ie, per disk). Tissue Preparation and DNA Bombardment About 150 to 200 mg of 7-day-old embryonic suspension cultures are placed in an empty, sterile 60 x 15 mm petri dish, and the dish is covered with plastic mesh. Tissue is bombarded with 1 or 2 shots per platelet with membrane burst pressure set at 1100 PSI and the chamber evacuated to a vacuum of 68.58 to 71.12 cm (27 to 28 inches) of mercury. The fabric is placed approximately 3.5 inches (7.62 cm) from the retention / detention screen. Selection of Transformed Embryos Transformed embryos are selected using either hygromycin (when the hygromycin phosphotransferase gene, HTP, was used as the selectable marker) or chlorosulforone (when the acetolactate synthase gene, ALS, was used as the selectable marker). Specifically, after bombardment, tissue is placed in fresh SB196 medium and cultured as described above. After 6 days of bombardment, the SB 196 is exchanged with fresh SB 196 containing either a 30 mg / l hygromycin selection agent or a 100 ng / ml chlorosulforone selection agent. The selection medium is renewed weekly. After 4 to 6 weeks of selection, transformed green tissue can be observed developing from non-transformed necrotic clusters. The isolated green tissue is removed and inoculated into multiwell platelets to generate new transformed and clonally propagated embryogenic suspension cultures. Somatic Somatic Embryo Regeneration in Plants To obtain whole plants from embryogenic suspension cultures, the tissue must be regenerated. Embryo Maturation Embryos can be cultured for 4 to 6 weeks at 26°C in SB196 under cool white (Phillips cool white Econowatt F40 / CW / RS / EW) and Agro (Phillips F40 Agro) fluorescent lamps (40 watts) on a photoperiod of 16 :8 h with light intensity from 90 to 120 pE / m 2s. After that time, the clusters are removed to a solid agar medium, SB166, for 1 to 2 weeks. Clusters are then subcultured on SB103 medium for 3 weeks. During this period, individual embryos can be removed from the pools and screened for changes in their fatty acid compositions, as described in Example 11. It should be noted that any detectable phenotype resulting from the expression of genes of interest could be screened at this time. phase. This would include (but is not limited to) changes in: the fatty acid profile, protein profile and content, carbohydrate content, growth rate, viability or the ability to grow normally in a soybean plant. Dissection and Embryo Germination Mature individual embryos can be dissected by placing them in a small, empty petri dish (35 x 10 mm) for about 4 to 7 days. Platelets are sealed with fiber tape (creating a small moisture chamber). The dissected embryos can be planted in SB71-4 medium where they are allowed to germinate under the same culture conditions described above. The germinated seedlings are removed from the germination medium and rinsed thoroughly with water and then planted in 24-cell Redi-Earth pack trays covered with clear plastic domes. After 2 weeks, the dome is removed and the plants are hardened for a further 1 week. If seedlings appear robust, they are transplanted into 25.4 cm (10”) Redi-Earth pots with up to three seedlings per pot. After 10 to 16 weeks, the mature seeds can be harvested, sliced ​​and analyzed for fatty acids as described above. Media Income SB 196 - FN Lite Liquid Proliferation Medium (per litre) MS FeEDTA - 100x Stock 1 10 ml MS Sulfate - 100x Stock 2 10 ml Halides FN Lite - 100x Stock 3 10 ml FN Lite P,B,Mo - 100x Stock 4 10 ml Vitamins B5 (1 ml / L) 1.0 ml 2,4-D (final concentration of 10 mg / L) 1.0 ml KNO3 2.83 g (NH 4 ) 2 ONLY 4 0.463 g Asparagine 1.0 g Sucrose (1%) 10g pH 5.8 Stock Solutions FN Lite Stock No. 1,000 ml 500 ml 1 - MS Fe EDTAWOx Stock Na 2 EDTA* 3.724 g 1.862 g FeSO 4 - 7H 2 O 2.784 g 1.392 g *Add first, dissolve in dark bottle while moving 2 - MS Sulfate 100x Stock MgSO 4 - 7H 2 O 37.0 g 18.5 g MnSO 4 - H 2 O 1.69 g 0.845 g ZnSO 4 - 7H 2 O 0.86 g 0.43 g CuSO 4 -5H 2 O 0.0025 g 0.00125 g 3 - Halides FN Lite 100x Stock CaCI 2 - 2H 2 O 30.0 g 15.0 g KI 0.083 g 0.0715 g CoCI 2 - 6H 2 O 0.0025 g 0.00125 g 4 - FN Lite P,B,Mo 100x Stock KH 2 DUST 4 18.5 g 9.25 g H3BO3 0.62 g 0.31 g Na 2 MoO 4 - 2H 2 O 0.025 g 0.0125 g Solid Medium SB1 (per liter) 1 pct. Of MS salts (Catalog 11117-066, Gibco / BRL) 1 ml of B5 vitamins 1,000 X Stock 31.5 g of sucrose 2 ml of 2,4-D (final concentration of 20 mg / l) pH 5.7 8 g deTC agar Solid medium SB 166 (per liter) 1 pct. Of MS salts (Catalog 11117-066, Gibco / BRL) 1 ml of B5 vitamins 1000 X Stock 60 g of maltose 750 mg of MgCI 2 hexahydrate 5 g activated charcoal pH 5.7 2 g of gelrite Solid medium SB 103 (per liter) 1 pct. MS salts (Catalog 11117-066, Gibco / BRL) 1 ml of B5 vitamins 1000 X Stock 60 g of maltose 750 mg of hexahydrated MgClz pH 5.7 2 g of gelrite Solid medium SB 71-4 (per liter) 1 bottle of Gamborg salt B5 with sucrose (Catalog 21153-036, Gibco / BRL) pH 5.7 5 g of TC agar 2.4 D Stock: previously obtained from Phytotech, Catalog D 295; concentration is 1 mg / ml Stock Vitamin B5 (per 100 ml; store aliquots at -20°C) 10 g myo-inositol 100 mg nicotinic acid 100 mg pyridoxine HCI 1 g thiamine * If solution does not dissolve quickly enough, apply a low level of heat through the stirring hot plate. Stock Chlorosulfuron 1 mg / ml in 0.01 N ammonium hydroxide | To induce somatic embryos, cotyledons, 3 to 5 mm long dissected from immature seeds with surface sterilized soybean culture A2872, can be cultured in light or in the dark at 26°C in an appropriate agar medium for 6 to 10 weeks. The somatic embryos, which produce secondary embryos, are then cut and planted in a suitable liquid medium. After repeated selection for the somatic embryo pools that have multiplied as before, the globular stage embryos, the suspensions are maintained as described below. Soybean embryogenic suspension cultures can be maintained in 35 ml of liquid medium on a gyratory shaker, 150 rpm, at 26°C, with fluorescent lights on a 16:8 hour day / night schedule. Cultures are subcultured every 2 weeks by inoculating about 35 mg of tissue in 35 ml of liquid medium. Soybean embryogenic suspension cultures can then be transformed by the particle gun bombardment method ( Klein et al., Nature (London) 327: 70-73 (1987 ); US 4,945,050 ). A DuPont Biolistic PDS1000 / HE instrument (helium retrofit) can be used for these transformations. A selectable marker gene that can be used to facilitate soybean transformation is a recombinant DNA construct composed of the 35S promoter of the Cauliflower Mosaic Virus (Odell et al., Nature 313: 810-812 (1985)), the gene for hygromycin phosphotransferase from plasmid pJR225 (from E. coli; Gritz et al., Gene 25: 179-188 (1983)) and the 3' region of the nopaline synthase gene from the T-DNA of the Ti plasmid from Agrobacterium tumefaciens. The seed expression cassette comprising the phaseolin 5' region, the fragment encoding the present polypeptide and the phaseolin 3' region can be isolated as a restriction fragment. This fragment can then be inserted into a unique restriction site on the vector carrying the marker gene. To 50 µl of a 60 mg / ml 1 µm gold particle suspension are added (in order): 5 µl DNA (1 pg / pl), 20 µl spermidine (0.1 M) and 50 µl of CaCl 2 (2.5M). The particle preparation is then shaken for 3 minutes, spun in a microcentrifuge for 10 seconds, and the supernatant is removed. The DNA-coated particles are then washed once in 400 µl of 70% ethanol and resuspended in 40 µl of anhydrous ethanol. The DNA / particle suspension can be sonicated three times for 1 second each. Five µl of DNA-coated gold particles are then loaded onto each macro carrier disc. About 300 to 400 mg of a 2-day-old suspension culture is placed in an empty 60 x 15 mm petri dish, and residual liquid is removed from the tissue with a pipette. For each transformation experiment, about 5 to 10 tissue platelets are typically bombarded. The membrane burst pressure is set to 7.58 x 10 3 kPa (1100 psi), and the chamber is evacuated to a vacuum of 71.12 cm (28 inches) of mercury. The fabric is placed approximately 7.62 cm (3.5 inches) away from the retention screen and blasted three times. After bombardment, the tissue can be divided in half and placed back in the liquid and cultured as described above. After 5 to 7 days of bombardment, the liquid medium can be exchanged with fresh medium, and 11 to 12 days after bombardment with fresh medium containing 50 mg / ml of hygromycin. The selective medium can be renewed weekly. After 7 to 8 weeks of bombardment, green transformed tissue can be observed developing from non-transformed necrotic embryogenic clusters. The isolated green tissue is removed and inoculated into individual flasks to generate new, transformed and clonally propagated embryogenic suspension cultures. Each newline can be treated as an independent transform event. These suspensions can then be subcultured and maintained as clusters of immature embryos or regenerated into whole plants by maturation and germination of individual somatic embryos. Example 14 Additional Codon-Optimized Delta-8 Desaturase Gene Modification FOR YARROWIA LIPOLYTICA The amino acid sequence of the codon-optimized synthetic D8S-3 gene in pDMW261 (Example 1) was corrected according to the amino acid sequence of the functional Euglena delta-8 desaturase (SEQ ID NO: 1 and 2). Using pDMW261 as a template and oligonucleotides ODMW404 (SEQ ID NO: 94) and D8-13R (SEQ ID NO: 36), the DNA fragment encoding the synthetic D8S-3 desaturase gene was amplified. The resulting PCR fragment was purified with Bio101's Geneclean kit and subsequently digested with Kpn1 and Not1 (primer CDMW404 introduced a KpnI site, while primer D8-13R introduced a NotI site). The KpnUNotl fragment (SEQ ID NO: 95) was cloned into KpnUNotl digest pKUNFmKF2 (Figure 5D; SEQ ID NO: 116) to produce pDMW277 (Figure 6A). Oligonucleotides YL521 (SEQ ID NO: 96) and YL522 (SEQ ID NO: 97), which were designed to amplify and correct the 5' end of the D8S-3 gene, were used as primers in another PCR reaction, where pDMW277 was used as the template. The primers introduced into the PCR fragment an Nco1 site and a Bglll site at its 5' and 3' ends, respectively. The 318 bp PCR product was purified with the Bio101 GeneClean kit and subsequently digested with Nco1 and Bglll. The digested fragment, with the 954 bp BglIlINnotl fragment of pDMW277, was used to exchange the NcoI / NotI fragment of pZF5T-PPC (Figure 6B; SEQ ID NO: 117) to form pDMW287 (Figure 6C). In addition, to correct the 5' end of the synthetic D8S-3 gene, this cloning reaction also placed the synthetic delta-8 desaturase gene under control of the Yarrowia lipolytica fructose-bisphosphate aldolase promoter containing a Yarrowia intron (FBAIN ;SEQ ID NO: 114; see WO 2005 / 049805). The first reaction in a final series of site-directed mutagenesis reactions was then performed on pDMW287. The first set of primers, YL525 (SEQ ID NO: 98) and YL526 (SEQ ID NO: 99), was designed to correct the F to S amino acid (position 50) of the synthetic gene D8S-3 in pDMW287. The plasmid resulting from this mutagenesis reaction then became the template for the next site-directed mutagenesis reaction with YL527 (SEQ ID NO: 100) and YL528 (SEQ ID NO: 101) as primers. These primers were designed to correct the F to S amino acid (position 67) of the D8S-3 gene and resulted in the creation of plasmid pDMW287 / YL527. To complete the sequence corrections within the second quarter of the gene, the following reactions were performed concurrently with mutations of the first quarter of the gene. Using pDMW287 as a template and oligonucleotides YL529 (SEQ ID NO: 102) and YL530 (SEQ ID NO: 103) as primers, an in vitro mutagenesis reaction was performed to correct the amino acid from C to W (position 177) of the synthetic gene D8S-3. The product (i.e., pDMW287 / Y529) of this mutagenesis reaction was used as the template in the following reaction using primers YL531 (SEQ ID NO: 104) and YL532 (SEQ ID NO: 105) to correct the amino acid from P to L (heading 213). The product of this reaction was called pDMW287 / YL529-31. Concomitant with mutations in the first and second quarters of the gene, reactions were similarly performed at the 3' end of the gene. Each subsequent mutagenesis reaction used the plasmid product of the preceding reaction. Primers YL533 (SEQ ID NO: 106) and YL534 (SEQ ID NO: 107) were used in pDMW287 to correct the amino acid from C to S (position 244) to create pDMW287 / YL533. Primers YL535 (SEQ ID NO: 108) and YL536 (SEQ ID NO: 109) were used to correct the amino acid from A to T (position 280) in the synthetic gene D8S-3 from pDMW287 / YL533 to form pDMW287 / YL533-5. Finally, the P of the amino acid at position 333 was corrected to S in the synthetic gene D8S-3 using pDMW287 / YL533-5 as the template and YL537 (SEQ ID NO: 110) and YL538 (SEQ ID NO: 110). 111) as primers. The resulting plasmid was named as pDMW287 / YL533-5-7. The Bglll / Xhol fragment of pDMW287 / YL529-31 and the Xhol / Not! of pDMW287 / YL533-5-7 was used to alter the Bglll INotl fragment of pDMW287 / YL257 to produce pDMW287F (Figure 6D) which contains the fully corrected synthetic delta-8 desaturase gene designated "D8SF" and shown in SEQ ID SEQ ID NO: 112. SEQ ID NO: 113 presents the amino acid sequence encoded by nucleotides 2 to 1270 of SEQ ID NO: 112, which is essentially the same as the sequence shown in SEQ ID NO: 2, except for an additional valine that accompanies the starting methionine. Example 15 Synthesis and Functional Expression of a CODON-Optimized Delta-9 Elongase Gene IN YARROWS UPOLYTICA To express the delta-9 elongase / delta-8 desaturase pathway in Yarrowia lipolytica, it was necessary to obtain an appropriate delta-9 elongase that could be co-expressed with the codon-optimized synthetic delta-8 desaturase of Example 14. of the Isochrysis galbana delta-9 elongase gene codon (GenBank Accession No. AF390174) was optimized for expression in Y. lipolytica. According to the Yarrowia codon usage pattern, the consensus sequence around the ATG translation initiation codon, and the general RNA stability rules (Guhaniyogi, G. and J. Brewer, Gene 265 (1-2 ): 11 to 23 (2001)), a codon-optimized delta-9 elongase gene was designed (SEQ ID NO: 118), based on the DNA sequence of Isochrysis galbana', SEQ ID NO: 119. In addition modification of the translation initiation site, 126 bp from the region of 792 bp encodings were modified and 123 codons were optimized. None of the modifications to the codon-optimized gene altered the amino acid sequence of the encoded protein (GenBank Accession No. AF390174; SEQ ID No.: 120). In Vitro Synthesis of a Codon Optimized Delta-9 Elongase Gene for Yarrowia The method used to synthesize the codon-optimized delta-9 elongase gene was the same as that used for the synthesis of the delta-8 desaturase gene (Example 1). First, eight pairs of oligonucleotides were designed to extend the entire length of the codon-optimized coding region of the / delta-9 elongase gene. galbana (e.g. IL3-1A, IL3-1B, IL3-2A, IL3-2B, IL3-3A, IL3-3B, IL3-4A, IL3-4B, IL3-5A, IL3-5B, IL3-6A, IL3 -6B, IL3-7A, IL3-7B, IL3-8A, IL3-8B, corresponding to SEQ ID NO: 121 to 136). Each pair of sense (A) and antisense (B) oligonucleotides was complementary, with the exception of a 4 bp overhang at each 5' end. Furthermore, primers IL3-1F, IL3-4R, IL3-5F and IL3-8R (SEQ ID NO: 137 to 140) also introduced NcoI, PstI and Not1 restriction sites, respectively, for subsequent subcloning. Each oligonucleotide (100 ng) was phosphorylated at 37°C for 1 hour in a 20 µl volume containing 50 mM Tris-HCI (pH 7.5), 10 mM MgCl 2 , 10 mM DTT, 0.5 mM spermidine, 0.5 mM ATP and 10 U T4 polynucleotide kinase. Each pair of sense and antisense oligonucleotides was mixed and annealed in a thermo-stirrer, using the following parameters: 95°C (2 min), 85°C (2 min), 65°C (15 min), 37°C, (15 min), 24°C (15 min) and 4°C (15 min). Therefore, IL3-1A (SEQ ID NO: 121) was annealed to IL3-1B (SEQ ID NO: 122) to produce the double-stranded product "IL3-1AB". Similarly, IL3-2A (SEQ ID NO: 123) was annealed to IL3-2B (SEQ ID NO: 124) to produce the double-stranded product "IL3-2AB", etc. Two separate groups of double-stranded looped oligonucleotides were then ligated together as shown below: Group 1 (comprising IL3-1AB, IL3-2AB, IL3-3AB and IL3-4AB); and Group 2 (comprising IL3-5AB, IL3-6AB, IL3-7AB and IL3-8AB). Each group of annealed oligonucleotides was mixed in a 20 µl volume with 10 U of T4 DNA ligase and the ligation reaction was incubated overnight at 16°C. The product of each ligation reaction was then used as a template to amplify the designated DNA fragment by PCR. Specifically, using the ligated "Group 1" mixture (i.e., IL3-1AB, IL3-2AB, IL3-3AB and IL3-4AB) as a template and the oligonucleotides IL3-1F and IL3-4R (SEQ ID NO: 137 and 138) as primers, the first part of the codon-optimized delta-9 elongase gene was amplified by PCR (as described in Example 1). The 417 bp PCR fragment was subcloned into the pGEM-T easy vector (Promega) to generate pT9(1-4). Using the ligated “Group 2” mixture (i.e., IL3-5AB, IL3-6AB, IL3-7AB and IL3-8AB) as the template and oligonucleotides IL3-5F and IL3-8R (SEQ ID NO: 139 and 140) as primers, the second part of the codon-optimized delta-9 elongase gene was similarly amplified by PCR and cloned into the pGEM-T easy vector to generate pT9(5-8). E. coli was transformed separately with pT9(1-4) and pT9(5-8), and plasmid DNA was isolated from ampicillin resistant transformants. The plasmid DNA was purified and digested with the appropriate restriction endonucleases to release the 417 bp NcolIPstI fragment of pT9(1-4) (SEQ ID NO: 141) and the 377 bp PstI / NotI fragment of pT9(5) -8) (SEQ ID NO: 142). These two fragments were then combined and directionally ligated together with NcoHNotl digested pZUF17 (SEQ ID NO: 143; Figure 7A), to generate pDMW237 (Figure 7B; SEQ ID NO: 144). The DNA sequence of the resulting synthetic gene delta-9 elongase ("lgD9e") in pDMW237 was exactly the same as the codon-optimized gene originally designed (ie, SEQ ID NO: 118) for Yarrowia. Lineage Generation of Y. lipolytica Y2031 (An Ura derivative of ATCC 20362) The Y2031 line was generated by integrating the chimeric TEF::YA12::Pex20 gene from the pKUNT2 plasmid ( Figure 7C ) into the Ura3 gene locus of Yarrowia lipolytica ATCC 20362 to thereby generate the Ura genotype of the Y2031 line. Specifically, the pKUNT2 plasmid contained the following components: Table 7 Description of Plasmid PKUNT2 (SEQ IP NO:145) ER Sites and Nucleotides within SEQ ID NO:145 Description of Fragment and Chimeric Gene Components Ascl / BsiWI (3225-3015) 784 per 5' part of the Yarrowia Ura3 gene (Gen Bank Accession No. AJ306421) Sphl / Pacl (5933-13) 516 bp 3' part of the Yarrowia Ura3 gene (GenBank Accession No. AJ306421) EcoRI / BsiWI (6380-8629 TEF::Y.Al2::Pex20, which comprises: - TEF: TEF promoter (GenBank Accession No. AF054508) - Y.A12: Yarrowia delta-12 desaturase gene (SEQ ID NO: 146; see also WO 2004 / 104167) - Pex20: Yarrowia Pex20 terminator sequence Pex20 gene (GenBank Accession No. AF054613) Plasmid pKUNT2 was digested with Ascl / Sphl, and then used for ATCC 20362 transformation of wild-type Y. lipolytica from according to the General Methods. Transforming cells were plated onto 5-fluoroorotic acid selection medium (“FOA”; also 5-fluorouracil-6-carboxylic acid monohydrate) platelets and kept at 30°C for 2 to 3 days. Specifically, the FOA selection medium comprised: 0.17% yeast nitrogen base (DIFCO Laboratories, Detroit, MI), no ammonium sulfate or amino acids, 2% glucose, 0.1% proline, 75 mg / l uracil, 75 mg / l uridine, 900 mg / l FOA (Zymo Research Corp., Orange, CA) and 20 g / l agar. FOA resistant colonies were picked and arrayed onto MM and MMU selection platelets. Colonies that could grow on MMU platelets but not MM platelets were selected as Ura- strains. Single colonies (5) of Ura- strains were then inoculated into liquid MMU at 30°C and shaken at 250 rpm / min for 2 days. Cells were collected by centrifugation, lipids were extracted and fatty acid methyl esters were prepared by transesterification and subsequently analyzed with a Hewlett-Packard 6890 GC. GC analyzes showed that there was about 45% LA in two Ura- strains (strains 2 and 3), compared to about 20% ATCC 20362 wild-type LA. Strain 2 transformant was designated as strain “Y2031”. Expression of the Codon-Optimized Delta-9 Elongase Gene in Y. upolytica Construct pDMW237 (comprising the chimeric FBAIN::lgD9e::Pex20 gene) was transformed into Yamowia Upolytica strain Y2031 as described in General Methods. Three transformants of Y2031 with pDMW237 were grown individually in MM medium for 2 days. Cells were collected by centrifugation, lipids were extracted and fatty acid methyl esters were prepared by transesterification and subsequently analyzed with a Hewlett-Packard 6890 GC instrument. The GC results showed that there were about 7.1%, 7.3% and 7.4% EDA produced in these transformants with pDMW237. These data demonstrated that synthetic igD9e could convert C18:2 to EDA. The “percent (%) substrate conversion” or “conversion efficiency” of the codon-optimized gene was determined to be about 13%, in 5 that the conversion efficiency was calculated according to the following formula: ([product] / [substrate+product])*100, where the 'product' includes the immediate product and all products in the pathway derived from it. This term refers to the efficiency with which the specific enzyme can convert substrate to product. Example 16 10 Expression of the Delta-9 Elongase / Delta-8 Desaturase Pathway to Produce DG LA in Yarrowia upolytica This Example describes DGLA biosynthesis and accumulation in Yarrowia Upolytica that has been transformed to express the delta-9 elongase / delta-8 desaturase pathway. Therefore, this required the co-synthesis of the codon-optimized synthetic delta-9 15 elongase (SEQ ID NO: 118; Example 15) and the codon-optimized synthetic delta-8 desaturase (SEQ ID NO: 112; Example 14) . Specifically, the Clal / Pad fragment comprising the chimeric FBAIN::D8SF::Pex16 gene from construct pDMW287F (Figure 6D) was introduced into the Clal / Pacl sites of pDMW237 (Figure 7B) to generate construct pDMW297 (Figure 7D). . Thus, plasmid pDMW297 contained the following components: Table 8 Description of Plasmid pDMW297(SEQ IP NO:148) ER Sites and Nucleotides within SEQ ID NO:148 Description of Fragment and Components of EcoRI / CIal Chimeric Genes ARS18 Sequence (GenBank Accession No. A17608) (9053-10448) ER Sites and Nucleotides within SEQ ID NO:148 Description of Fragment and Components of Clal / Pacl(1-2590) FBAIN::A8S::Pex16 Chimeric Genes, which comprises: - FBAIN: FBAIN promoter (SEQ ID NO:114 ) - A8S: codon-optimized delta-8 desaturase gene (SEQ ID NO:112), derived from Euglena gracilis (GenBank Accession No. AF139720) - Pex16: Pex16 terminator sequence from Yarrowia pex16 gene (GenBank Accession No. U75433) Pacl / Sall (2590-4082) - Yarrowia Ura3 gene (GenBank Accession No. AJ306421) Sall / BsiWI (4082-6257) FBAIN::A9ES::Pex120, comprising: - FBAIN: FBAIN promoter (SEQ ID NO: 114) - A9ES: codon-optimized delta-9 elongase gene (SEQ ID NO: 118), derived from Isochrysis galbana (GenBank Accession No. 390174) - Pex20: Pex20 terminator sequence from Yarrowia Pex20 gene (GenBank Accession No. AF054613) Plasmid pDMW297 was then used for transformation of the strain Y2031 (sample 15) according to the general methods. Transforming cells were plated onto MM selection medium plates and kept at 30°C for 2 to 3 days. A total of eight transformants grown on the MM plates were chosen and replated on fresh MM plates. Once developed, these strains were individually inoculated into liquid MM at 30°C and shaken at 250 rpm / min for 2 days. Cells were collected by centrifugation, lipids were extracted and fatty acid methyl esters were prepared by transesterification and analyzed in 10 sequences with a Hewlett-Packard 6890 GC instrument. GC analyzes showed that DGLA was produced in all transformants analyzed. One strain produced about 3.2%, 4 strains produced 4.3 - 4.5%, two strains produced 5.5 - 5.8%, and one strain produced 6.4% DGLA (hereinafter referred to as the lineage "Y0489"). The "percent (%) substrate conversion" of the codon-optimized D8SF gene in the Y0489 lineage was determined to be equal to 75% (using the formula of Example 15). It will be obvious to one skilled in the art that other chimeric genes could be co-expressed with the D8SF and lgD9e genes in genetically engineered Yarrowia to allow the production of various other PUFAs. For example, in addition to the codon-optimized delta-9 elongase and delta-8 desaturase genes, it was possible to readily express: (1) a delta-15 desaturase to enable ETA production; (2) a delta-5 desaturase to allow production of ARA; (3) a delta-17 desaturase to allow ETA production; (4) a delta-5 desaturase and a delta-17 desaturase to allow the production of EPA; (6) a delta-5 desaturase, a delta-17 desaturase and a C20 / C22 elongase to allow production of DPA; or (7) a delta-5 desaturase, a delta-17 desaturase, a C20 / C22 elongase and a delta-4 desaturase to allow the production of DHA (Figure 9). Example 17 Cloning of Delta-8 Desaturase from Euglena gracilis into a Soybean Expression Vector and Co-Expression With an Elongase from Isochrysis GALBANA The gene for Isochrysis galbana elongase was amplified from pDMW237 (Figure 7B; SEQ ID NO:144) using primers olGsel1-1 (SEQ ID NO:149) and olGsel1-2 (SEQ ID NO:150) which were designed to insert Notl restriction enzyme sites at both ends of the elongase. The resulting PCR fragment was digested with NotI and cloned into the NotI site of pKR72 to obtain pKR607. Plasmid pKR680 was digested with BsiWI and the fragment containing Eg5 (SEQ ID NO:1) was cloned into the BsiWI site of pKR607 to thereby obtain pKR682. Thus, delta-8 desaturase (Eg5; SEQ ID NO:1) could be co-expressed with elongase from Isochrysis galbana behind strong seed-specific promoters. A map of pKR682 is shown in Figure 8A. Example 18 Assembly of EPA Biosynthetic Pathway Genes With Delta-8 Desaturase from Euglena gracilis and Elongase from Isochrysis galbana For Expression in Somatic Soybean Embryos and Soybean Seeds A soybean expression vector (pKR786) that contains the delta- 8 desaturase from Euglena gracilis, delta-9 elongase from Isochrysis galbana and delta-5 desaturase from Mortierella alpina (all under the control of strong seed-specific promoters) was constructed as follows. Through a number of subcloning steps, a DNA sequence (SEQ ID NO: 151) was effectively added into the Smal site of the pKR287 vector (WO 2004 / 071467 A2) to produce pKR767. In this way, an Sbfl restriction site was added to the 3' end of the leg1A transcription terminator of the Gy1 / Mad5 / legA2 cassette. The Asci fragment of pKR682 was cloned into the Asci site of pKR277 (WO 2004 / 071467 A2) to produce pKR769. The Gy1 / Mad5 / legA2 cassette was released from pKR767 via Sbfl digestion and the resulting fragment was cloned into the Sbfl site of pKR769 to produce pKR786. A map of pKR786 is shown in Figure 8B. Example 19 Cloning of Fusarium Delta-15 Desaturase into a Soybean Expression Vector and Co-Expression With EPA Biosynthetic Genes (Delta-15 Desaturase, Delta-17 Desaturase) The Kti3.Fm A15 desaturase ORF:Kti3 promoter terminator cassette was released from plasmid pKR578 (WO 2005 / 047479) by BsiWI digestion and cloned into the BsiWI site of plasmid pKR226 (WO 2004 / 071467 A2), containing the gene of ALS for selection, the T7prom / hpt / T7term cassette and the bacterial ori region, to produce pKR667. Plasmid pKR271 was digested with PstI and the fragment containing the delta-17 desaturase from Saprolegnia diclina was cloned into the Sbfl site of pKR667 to produce pKR669. In this way, the delta-15 desaturase could be co-expressed with the delta-17 desaturase from Saprolegnia diclina behind strong seed-specific promoters. A map of pKR669 is shown in Figure 8C. Example 20 Analysis of Somatic Soybean Embryos Containing Euglena gracilis Delta-8 Desaturase and Mortierella alpina Elongase Genes (PKR681) Mature somatic soybean embryos are a good model for zygotic embryos. While in the globular embryo state in liquid culture, somatic soybean embryos contained very low amounts of triacylglycerol or storage proteins typical of mature, zygotic soybean embryos. At this stage of development, the ratio of total triacylglyceride to total polar lipid (phospholipids and glycolipids) is about 1:4, as is typical of zygotic soybean embryos at the stage of development from which the embryo culture somatic was initiated. Also at the globular stage, the mRNAs for the prominent seed proteins, the p-conglycinin α'-subunit, the 3-inhibitor kunitz trypsin, and seed lectin are essentially absent. With and transfer to hormone-free media to allow differentiation to the maturation somatic embryo state, triacylglycerol becomes the most abundant class of lipid. Furthermore, mRNAs for α'-conglycinin α'-subunit, kunitz trypsin-3 inhibitor, and seed lectin become very abundant messages in the total mRNA population. On this basis, the soybean somatic embryo system behaves very analogously to in vivo maturing soybean zygotic embryos, and is therefore a good and rapid model system for analyzing the phenotypic effects of modifying the expression of fatty acid genes on biosynthetic pathway (example 3 in WO 02 / 00904). Even more importantly, the model system is also predictive of the fatty acid composition of the seeds of the plants derived from the transgenic embryos. Somatic embryos from transgenic soybeans containing the constructs described above were analyzed in a similar manner. For this, fatty acid methyl esters were prepared from single mature soybean somatic embryos by means of transesterification. The embryos were placed in a flask containing 50 µl of trimethylsulfonium hydroxide (TMSH) and 0.5 ml of hexane, and incubated for 30 minutes at room temperature under agitation. The fatty acid methyl esters (5 µl injected from the hexane layer) are separated and quantified using a Hewlett-Packard 6890 gas chromatograph equipped with an Omegawax 320 fused silica capillary column (catalog 24152, Supelco Inc.). The oven temperature was programmed to hold at 220°C for 2.7 minutes, increase to 240°C at 20°C / min and then hold for another 2.3 minutes. The carrier gas was supplied by a Whatman hydrogen generator. Retention times were compared to those for methyl esters of commercially available standards (Catalog U-99-A, Nu-Chek Prep, Inc.). Routinely, six to ten embryos per event were analyzed by GC, using the methodology described above. More specifically, the fatty acid profiles of embryos for about six lines containing pKR681 are shown in Table 9. The best line (i.e., 1618-1-1-1) had embryos with an average DGLA content of 8, 9% and an average ETA content of 3.1%. For strains 1618-1-8-1, 1618-3-6-1 and 1618-4-1-1, only elongase seemed to work. The best elongase strain (ie, 1618-4-1-1) had embryos with an average EDA content of 10.6% and an average EtrA content of 6.5%. Percent elongation, percent desaturation, and calculated elongation and desaturation rates are shown in Table 10. In the 1618-1-1-1 strain, delta-8 desaturase converts an average of 76.3% of the fatty acids C20 elongated in the product with the best embryo conversion of 82.1% in the product. Delta-8 desaturase appears to utilize EDA and EtrA equally well, since the ratio of their respective desaturation percentages is about 1.0. In strain 1618-4-1-1, elongase from Mortierella alpina converts an average of 23% of the C18 fatty acids into product with the best embryo conversion of 30.2% into product. Elongase appears to have a slight preference for ALA, since the ratio of their respective elongation percentages is about 0.6. Elongase-only expression in these strains likely resulted from construct fragmentation during the transformation procedure or due to the effects of positional insertion differentially affecting delta-8 expression. Table 9 Accumulation of Long Chain PUFAs in Lineages Transformed with PKR681 Lineage 16:0 18:0 18:1 LA GLA ALA EDA DGLA EtrA ETA 1618-1-1-1 13.9 6.8 7.1 40.2 0.0 14.4 3.0 10.2 1, 1 3.3 -2 14.5 10.0 6.2 38.9 0.0 11.0 4.1 10.5 1.2 3.0 -3 14.1 4.9 4.7 42.2 0.0 21.5 2.0 7.2 1.0 2.5 -4 14.1 7.1 6.2 42.7 0.0 13.3 3.3 9.1 1.1 2.8 Lineage 16:0 18:0 18:1 LA GLA ALA EDA DGLA EtrA ETA -5 12.0 5.0 5.8 46.3 0.0 16.0 2.2 8.1 1.0 3.2 - 6 12.1 4.7 5.7 42.0 0.0 20.9 1.7 8.5 0.9 3.5 Average 13.5 6.4 6.0 42.1 0.0 16.2 2.7 8.9 1.0- 3.1 1618-1-2-1 11.7 4.3 4.6 46.8 0.0 17.4 3.9 5.7 2.4 3.1 -2 12.2 6.0 4.7 45.5 0.0 14.9 5.1 5.8 2.9 2.9 -3 12.7 5.0 7.1 44.7 0.0 17 .5 4.2 4.1 2.4 2.3 -4 12.4 6.3 6.8 43.0 0.0 13.9 6.6 4.9 3.9 2.3 -5 13, 4 8.7 5.2 39.6 0.0 13.2 6.3 7.2 3.2 3.1 -6 12.8 5.5 6.2 45.7 0.0 15.6 4, 3 5.2 2.2 2.5 Average 12.5 6.0 5.8 44.2 0.0 15.4 5.1 5.5 2.8 2.7 1618-1-8-1 8, 7 3.5 6.7 53.3 0.0 17.2 6.9 0.0 3.6 0.0 -2 9.2 2.9 12.0 49.0 0.0 18.9 4, 7 0.0 3.3 0.0 -3 11.2 2.8 7.7 48.6 0.0 22.4 4.1 0.0 3.2 0.0 -4 12.0 3.6 13.6 46.7 0.0 16.0 4.8 0.0 3.2 0.0 -5 9.1 3.6 5.0 52.6 0.0 16.5 8.5 0.0 4.8 0.0 -6 9.3 2.8 12.7 47.2 0.0 20.0 4.6 0.0 3.4 0.0 Average 9.9 3.2 9.6 49, 6 0.0 18.5 5.6 0.0 3.6 0.0 1618-3-6-1 11.8 2.4 8.3 42.1 0.0 28.2 3.3 0.0 3.9 0.0 -2 10.6 4.2 12.8 43.7 0.0 17.6 6.7 0.0 4.3 0.0 -3 10.3 4.9 5.6 45 .7 0.0 18.4 8.7 0.0 6, 3 0.0 -4 11.8 5.2 21.2 39.5 0.0 15.2 4.4 0.0 2.8 0.0 -5 10.3 3.0 9.2 47.8 0.0 21.5 4.7 0.0 3.5 0.0 -6 9.4 2.5 9.4 47.9 0.0 23.2 4.0 0.-0 3.7 0, 0 Lineage 16:0 18:0 18:1 LA GLA ALA EDA DGLA EtrA ETA Average 10.7 3.7 11.1 44.4 0.0 20.7 5.3 0.0 4.1 0.0 1618-4- 1-1 15.4 9.2 6.5 38.1 0.0 9.9 13.8 o.o 7.0 0.0 -2 11.1 5.6 5.7 43.3 0.0 16, 8 10.2 0.0 7.4 0.0 -3 10.5 5.0 6.6 45.4 0.0 15.4 10.1 0.0 6.9 0.0 -4 10.2 5.8 6.5 45.1 0.0 12.9 12.3 0.0 7.2 0.0 -5 11.4 4.4 10.1 45.3 0.0 16.1 7.4 0.0 5.2 0.0 -6 10.7 5.2 13.6 42.9 0.0 12.6 9.6 0.0 5.3 0.0 Average 11.5 5.9 8, 2 43.4 o.o 14.0 10.6 0.0 6.5 0.0 The fatty acid compositions listed in Table 9 are expressed as a percentage by weight. 16:0 = palmitic acid, 18:0 = stearic acid, 18:1 = oleic acid, l_A = linoleic acid, G!_A = y-linoleic acid, ALA = alpha-linolenic acid, EDA = eicosadienoic acid, DGLA = dihomo-γ-linoleic acid, EtrA = eicosatrienoic acid, ETA = eicosa-tetraenoic acid. Table 10 Comparison of Percentage of Desaturation and Percentage of Elongation in Lineages Transformed with pKR681 Cis Lineage %Elong C 2 the % of desat. of delta-8 LA % of Elong ALA % of Elong Ratio (LA / ALA) Elong EDA % of desat. of delta-8 EtrA % of desat. of delta-8 Ratio (EDA / EtrA) desat. of delta-8 1618-1-1-1 24.4 76.6 24.7 23.5 1.1 77.1 75.0 1.0 -2 27.3 71.9 27.2 27.7 1.0 72.1 71.1 1.0 -3 16.6 76.4 17.9 14.0 1.3 78.1 72.0 1.1 -4 22.5 73.3 22.4 22.7 1.0 73.5 72.6 1.0 Elong C % Cis Lineage 20 % of desat. of delta-8 LA % of Elong ALA % of Elong Ratio (LA / ALA) Elong EDA % of desat. of delta-8 EtrA % of desat. of delta-8 Ratio (EDA / EtrA) desat. from delta-8 -5 18.9 77.7 18.3 20.7 0.9 78.4 76.0 1.0 -6 18.8 82.1 19.6 17.4 1.1 83.1 79, 9 1.0 Average 21.4 76.3 21.7 21.0 1.1 77.0 74.4 1.0 1618-1-2-1 19.1 58.1 17.0 24.0 0, 7 59.0 56.6 1.0 -2 21.7 52.2 19.3 28.0 0.7 53.2 50.4 1.1 -3 17.3 49.2 15.8 21.0 0.8 49.5 48.5 1.0 -4 23.7 40.6 21.0 30.8 0.7 42.6 36.9 1.2 -5 27.3 51.8 25.5 32 .2 0.8 53.3 48.6 1.1 -6 18.9 54.1 17.3 23.1 0.7 54.6 53.1 1.0 Average 21.3 51.0 19.3 26.5 0.7 52.0 49.0 1.1 1618-1-8-1 13.0 11.5 17.5 0.7 -2 10.6 8.8 14.8 0.6 -3 9.4 7.9 12.5 0.6 -4 11.4 9.3 16.8 0.6 -5 16.2 13.9 22.7 0.6 -6 10.7 9.0 14, 6 0.6 Average 11.9 10.1 16.5 0.6 1618-3-6-1 9.3 7.4 12.0 0.6 -2 15.2 13.3 19.5 0.7 -3 19.0 15.9 25.6 0.6 lineage c 18 % Elong C20 % Desat. of delta-8 LA % of Elong ALA % of Elong Ratio (LA / ALA) Elong EDA % of desat. of delta-8 EtrA % of desat. of delta-8 Ratio (EDA / EtrA) desat. of delta-8 -4 11.6 10.0 15.6 0.6 -5 10.6 9.0 14.0 0.6 -6 9.8 7.7 13.9 0.6 Average 12.6 10.5 16.8 0.6 1618-4-1-1 30.2 26.6 41.4 0.6 -2 22.6 19.0 30.5 0.6 -3 21.9 18.2 31.0 0.6 -4 25.2 21.5 35.8 0.6 -5 17.1 14.1 24.4 0.6 -6 21.1 18.3 29.6 0.6 Average 23 .0 19.6 32.1 0.6 The elongation percentage of (% of elong, of Cis) in the Table 10 was calculated by dividing the sum of the weight percentage for EDA, DGLA, EtrA and ETA (Table 9) by the sum of the weight percentage for LA, ALA, EDA, DGLA, EtrA and ETA (Table 9) and by multiplying by 100 to express as a 5 percentage. The percentage of A8 C desaturation 2 o (% of A8 desat of C 20 , Table 10) was calculated by dividing the sum of the weight percentage for DGLA and ETA (Table 9) by adding the percentage by weight for EDA, DGLA, EtrA and ETA (Table 9) and multiplying by 100 to express as a percentage. The individual elongations (percentage of LA elong. or percentage of 10 ALA elong.) or A8 desaturations (EDA percent of A8 desat or EtrA percentage of A8 desat) shown in Table 10 were calculated from a similar way but only when using the w-6 substrates / products or the u>-3 substrates / products for each. The elongation ratio for LA and ALA was obtained by dividing the percentage of LA elongation by the percentage of ALA elongation. Similarly, the delta-8 desaturation ratio was obtained by dividing the percentage of delta-8 desaturation of EDA by the percentage of delta-8 desaturation of EtrA. Example 21 Analysis of Somatic Soybean Embryos Containing Genes for Delta-8 Desaturase from Euglena gracilis and Elongase from Isochrysis galbana (PKR682) Embryo fatty acid profiles for nine strains containing pKR682 are shown in Table 11. Percent elongation, percent desaturation, and calculated elongation and desaturation ratios are shown in Table 12. The best strain (1619-6 -7) had embryos with an average DGLA content of 21.8% and an average ETA content of 4.1%. As can be seen in Table 12, in this strain, delta-8 desaturase converts an average of 91.6% of C fatty acids 2 elongation in the product and, in the best embryo (1619-6-7-1), 95.1% of the elongated fatty acids are converted to the product. With regard to pKR681, the delta-8 desaturase appears to utilize EDA and EtrA equally well with the ratio between their respective percentages of desaturation being around 1.0 (Table 12). In these strains, the average percentage conversion of Cis fatty acids to C fatty acids 2 o ranges from 40.0% to 49.5%. As can be seen with pKR681, there are strains (1619-6-4,1619-8-4) where only elongase is functioning (Table 11). Again, this is likely due to positioning effects or DNA fragmentation. In the lines where the delta-8 desaturase is not working, the best elongase line (1619-6-4) had embryos with an average EDA content of 24.1% and an average ETrA content of 8.7%. The best analyzed embryo had 27.4% of EDA and 10.3% of ETrA. The average elongation in this lineage is 49.5% with the best embryo (1619-6-4-6) having an elongation of 58.9% (Table 12). In these strains, elongase appears to have no preference over LA or ALA, since the ratio of their respective elongation percentages is about 1.0. Interestingly, in strains that also express delta-8 desaturase, there appears to be a slight preference of elongase over LA and the mean elongation ratio is as high as 2.3 in the 1617-16-2-7 strain. In many of the strains, a small amount of the fatty acid that follows a retention time identical to GLA is present when the delta-8 desaturase is functioning well. Table 11 Accumulation of Long Chain PUFAs in Lines Transformed with PKR682 16:0 18:0 18:1 LA GLA ALA EDA DGLA EtrA ETA 1617-16-2-7 20.1 2.7 5.9 18.0 2.0 12.8 5.1 26.5 2.4 4.4 -8 19.3 1.3 5.0 22.7 1.7 23.4 3.7 16.8 1.6 4.5 -9 20.4 2.4 4.7 13.7 2.3 15, 3 5.2 26.2 3.5 6.3 -10 17.0 1.7 6.2 19.9 1.7 27.0 2.5 17.4 1.1 5.5 -11 16.4 1.3 5 .1 21.5 3.3 28.2 2.8 15.2 2.1 4.1 -12 26.7 2.4 6.1 0.0 4.1 20.2 6.5 26.9 2.2 5.0 -13 17.5 1.5 5.8 21.6 2.6 20.3 3.9 19.9 1.8 5.1 -14 20.2 2.4 8.9 24.6 1.6 17.9 4 .3 14.7 1.4 4.2 Average 19.7 2.0 6.0 17.7 2.4 20.6 4.2 20.4 2.0 4.9 1619-6-4-1 18.5 1.7 9.4 25.0 0.0 8.2 27.4 0.0 9.7 0.0 -2 14.5 2.0 15.9 26.8 0.0 7.8 24.5 0, 0 8.5 o.o -3 23.6 3.8 12.2 19.3 0.0 8.7 23.8 0.0 8.8 0.0 16:0 18:0 18:1 LA GLA ALA EDA DGLA EtrA ETA -4 15.5 1.2 12.6 34.5 0.0 14.7 15.4 0.0 6.0 o.o -5 15, 2 1.6 15.7 25.5 o.o 6.7 26.4 0.0 8.9 0.0 -6 15.8 2.2 18.0 19.1 0.0 7.2 27, 4 0.0 10.3 0.0 Average 17.2 2.1 14.0 25.0 0.0 8.9 24.1 0.0 8.7 o.o 1619-6-5-1 22.1 2 ,1 6.2 23.9 3.9 10.5 4.2 21.1 1.3 4.8 -2 17.4 1.6 9.5 32.3 1.5 11.0 3.0 18 ,0 0.7 4.6 -3 17.5 2.6 9.9 32.9 0.5 11.3 5.8 15.3 0.6 3.3 -4 17.2 2.0 12, 1 29.5 0.6 10.6 6.0 13.9 2.8 4.7 -5 24.2 3.1 7.1 25.4 2.0 10.7 5.2 16.8 1, 9 3.4 -6 17.9 1.6 5.9 30.8 2.4 12.3 5.7 18.2 1.6 3.7 Average 19.4 2.2 8.5 29.2 1 .8 11.1 5.0 17.2 1.5 4.1 1619-6-7-1 19.1 1.7 4.8 32.0 1.1 21.7 1.0 16.3 0, 0 2.3 -2 19.0 1.3 5.0 40.1 1.2 17.9 1.4 11.9 0.2 2.0 -3 17.8 1.2 6.2 26.6 2, 0 13.1 2.1 24.6 0.4 5.9 -4 19.4 1.3 8.1 29.4 1.2 12.6 3.0 20.5 0.5 4.1 -5 19.9 1.4 9.2 19.6 3.1 8.8 2.1 29.9 0.5 5.4 -6 20.1 1.6 6.9 25.0 2.9 8.4 2, 5 27.6 0.4 4.6 Average 19.2 1.4 6.7 28.8 1.9 13.7 2.0 21.8 0.3 4.1 1619-7-3-1 15, 4 1.9 9.4 34.7 0.6 12.1 11.9 6.7 4.3 3.0 -2 15.2 1.5 9.6 37.4 0.0 17.0 9.9 3.7 3.6 1.9 -3 17.0 3.0 14.5 26.6 0.5 9.9 11.0 10.5 3.1 3.9 -4 18.5 3 .4 8.6 17.7 1.3 4.2 21.7 16.9 4.0 3.7 -5 16.5 2.4 10.2 25.8 0.8 7.0 15.1 12 .6 4.7 5.0 16:0 18:0 18:1 LA GLA ALA EDA DGLA EtrA ETA -6 16.9 2.2 10.3 24.4 0.4 6.8 22.7 6.3 7.3 2.6 Average 16 .6 2.4 10.4 27.8 0.6 9.5 15.4 9.5 4.5 3.4 1619-7-7-1 21.2 1.5 12.8 17.6 1, 3 8.8 6.8 23.1 2.1 4.8 -2 15.1 1.2 19.9 27.7 0.7 11.2 7.4 12.1 1.7 3.0 -3 17, 4 2.1 16.4 23.8 0.6 9.1 10.4 15.2 1.9 3.2 -4 17.2 1.5 18.3 21.4 0.9 9.2 9.3 16 .6 1.8 3.9 -5 16.4 1.0 13.4 24.2 1.2 15.9 6.5 16.2 2.1 3.3 -6 20.3 2.3 7.5 19 .5 1.3 9.5 8.9 22.9 2.4 5.1 Average 17.9 1.6 14.7 22.4 1.0 10.60 8.2 17.7 2.0 3, 9 1619-7-8-1 19.2 1.8 5.7 21.7 2.1 11.1 12.1 17.5 4.0 4.9 -2 15.0 1.1 10.6 28 .5 1.0 16.2 10.3 10.8 3.5 3.1 -3 17.0 1.6 11.3 20.6 1.2 8.6 12.4 17.8 4.3 5 .2 -4 16.5 1.5 10.3 25.4 1.0 13.6 11.1 13.4 3.3 3.8 -5 16.0 1.3 10.0 29.0 0, 8 12.6 4.7 19.0 1.2 5.5 -6 15.6 1.6 12.2 28.7 1.0 9.9 9.4 15.6 1.7 4.6 Average 16 ,6 1.5 10.0 25.6 1.2 12.0 10.0 15.7 3.0 4.5 1619-8-1-1 20.4 1.7 5.8 24.4 2, 7 15.0 4.3 20.3 1.5 3.9 -2 17.2 1.9 14.5 24.2 0.0 9.3 5.0 20.9 1.1 5.9 -3 16.4 1.8 13.0 23.5 1.4 11.6 6.9 19.9 1.3 4.1 -4 17.2 1.5 15.3 22.2 1.1 7.2 7.7 21.5 1.2 4.9 -5 19.4 1.3 9.9 21.7 2.8 13.8 5.1 20.2 1.7 4.1 -6 17.9 1.3 10.5 23.4 1.5 9.9 6.5 22.8 1.4 4.8 Average 18.1 1.6 11.5 23.3 1, 6 11.1 5.9 20.9 1.4 4.6 16:0 18:0 18:1 LA GLA ALA EDA DGLA EtrA ETA 1619-8-4-1 15.7 1.1 6.7 50.5 0.0 18.1 6.2 0.0 1.8 0.0 -2 15.0 1.8 8.2 38.6 0.0 28.4 5.8 0.0 2.2 0.0 -3 18.1 2.9 7.2 35.6 0 .0 32.2 2.7 0.0 1.3 0.0 -4 18.0 2.7 9.7 40.7 0.0 18.2 8.7 0.0 1.9 0.0 - 5 16.0 1.5 6.9 50.4 0.0 20.8 3.0 0.0 1.4 0.0 -6 15.3 0.9 7.7 50.8 0.0 20, 6 3.6 0.0 1.2 0.0 Average 16.4 1.8 7.7 44.4 0.0 23.00 5.0 0.0 1.6 0.0 The compositions listed in Table 11 are expressed as percentage by weight of fatty acid. 16:0 = palmitic acid, 18:0 = stearic acid, 18:1 = oleic acid, LA = linoleic acid, GLA = y-linoleic acid, ALA = alpha-linolenic acid, EDA = 5 eicosadienoic acid, DGLA = dihomo-γ-linoleic acid, EtrA = eicosatrienoic acid, ETA = eicosatetraenoic acid. Table 12 Comparison of Percentage of Desaturation and Percentage of Elongation in Lineages Transformed with pKR682 Strain Ci8 % Elong C20 % desat. of delta-8 LA % of Elong ALA % of Elong Ratio (LA / ALA) of Elong EDA % of desat. of delta-8 EtrA % of desat. of delta-8 Desat. of delta-8 1617-16-2- 55.5 80.5 63.8 34.7 1.8 83.9 65.0 1.3 7 36.6 80.2 47.4 20.8 2.3 82.1 73.6 1.1 -8 58.8 78.8 69.6 39.3 1.8 83.4 64.1 1.3 -9 36.0 86.6 49.9 19.5 2.6 87.5 83.9 1.0 -10 32.8 79.6 45.6 18.1 2.5 84.3 66.1 1.3 lineage c 18 % of Elong C 20 % of desat. of delta-8 LA % of Elong ALA % of Elong Ratio (LA / ALA) of Elong EDA %desat. of delta-8 EtrA % of desat. of delta-8 Desat. from delta-8 -11 66.7 78.6 100.0 26.1 3.8 80.6 69.5 1.2 -12 42.3 81.3 52.4 25.4 2.1 83.5 74.0 1.1 -13 36.6 76.7 43.5 23.8 1.8 77.4 74.4 1.0 -14 Average 45.7 80.3 59.0 26.0 2.3 82.8 71.3 1.2 1619-6-4-1 52.8 52.3 54.2 1.0 -2 48.8 47.7 52.1 0.9 -3 53.8 55.2 50, 3 1.1 -4 30.3 30.8 29.1 1.1 -5 52.3 50.9 57.0 0.9 -6 58.9 58.9 58.9 1.0 Average 49.5 49.3 50.3 1.0 1619-6-5-1 47.6 82.5 51.3 36.7 1.4 83.4 78.5 1.1 -2 37.8 85.8 39, 4 32.3 1.2 85.5 86.9 1.0 -3 36.0 74.5 39.0 25.5 1.5 72.6 84.7 0.9 -4 40.6 67.7 40.2 41.7 1.0 69.6 62.5 1.1 -5 43.1 74.0 46.4 33.2 1.4 76.3 64.6 1.2 -6 40.4 74 ,9 43.7 30.1 1.5 76.0 70.1 1.1 Average 40.9 76.6 43.3 33.2 1.3 77.3 74.6 1.0 1619-6-7-1 26.8 95.1 35.1 9.6 3.6 94.4 100.0 0.9 Cia strain % Elong C20 % desat. of delta-8 LA % of Elong ALA % of Elong Ratio (LA / ALA) of Elong EDA % of desat. of delta-8 EtrA % of desat. of delta-8 Desat. from delta-8 -2 21.2 89.5 25.0 11.1 2.3 89.4 90.1 1.0 -3 45.4 92.4 50.1 32.4 1.5 92.0 94, 1 1.0 -4 40.1 87.6 44.4 26.5 1.7 87.2 89.9 1.0 -5 57.2 93.2 62.0 40.1 1.5 93.4 92.3 1 .0 -6 51.2 91.9 54.6 37.4 1.5 91.8 92.9 1.0 Average 40.3 91.6 45.2 26.2 2.0 91.4 93.2 1.0 1619-7-3-1 35.6 37.6 34.9 37.6 0.9 36.0 41.5 0.9 -2 26.1 29.4 26.8 24.5 1.1 27 .3 34.6 0.8 -3 43.9 50.4 44.7 41.1 1.1 48.8 55.2 0.9 -4 67.8 44.5 68.5 64.6 1.1 43 .8 48.1 0.9 -5 53.3 47.0 51.8 58.0 0.9 45.4 51.6 0.9 -6 55.5 23.0 54.3 59.2 0.9 21 .8 26.5 0.8 Average 47.0 38.7 46.8 47.6 1.0 37.2 42.9 0.9 1619-7-7-1 58.3 76.7 63.0 44.0 1.4 77.1 -2 38.4 62.4 41.3 29.8 1.4 61.9 86.9 1.0 -3 48.3 60.0 51.8 36.0 1.4 59.3 84.7 0 .9 -4 50.7 64.9 54.6 38.2 1.4 64.2 62.5 1.1 -5 41.1 69.5 48.3 25.4 1.9 76.3 64, 6 1.2 -6 57.5 71.1 62.0 44.0 1.4 76.0 70.1 1.1 Average 49.0 67.2 53.5 36.2 1.5 77.3 76.6 1.0 Lineage Cj8% of Elong C 2 the % of desat. of delta-8 LA % of Elong ALA % of Elong Ratio (LA / ALA) of Elong EDA % of desat. of delta-8 EtrA % of desat. of delta-8 Desat. of delta-8 1619-7-8-1 54.0 58.2 57.7 44.4 1.3 59.2 54.9 1.1 -2 38.2 50.1 42.6 28.8 1.5 51.2 46.6 1.1 -3 57.6 57.9 59.4 52.2 1.1 58.9 54.6 1.1 -4 44.8 54.5 49.1 34.5 1 ,4 54.8 53.5 1.0 -5 42.2 80.7 45.0 34.7 1.3 80.3 82.1 1.0 -6 44.7 64.6 46.5 38, 8 1.2 62.4 73.0 0.9 Average 46.9 61.0 50.1 38.9 1.3 61.1 60.8 1.0 1619-8-1-1 43.2 80, 7 50.2 26.4 1.9 82.6 72.1 1.1 -2 49.5 81.5 51.7 42.7 1.2 80.7 84.7 1.0 -3 47.9 74.5 53.3 31.8 1.7 74.2 76.0 1.0 -4 54.6 74.1 56.8 45.8 1.2 73.7 79.6 0.9 -5 46 ,6 78.3 53.8 29.4 1.8 80.0 70.9 1.1 -6 51.5 77.8 55.6 38.3 1.5 77.8 77.9 1.0 Average 48.9 77.9 53.6 35.7 1.6 78.2 76.9 1.0 1619-8-4-1 10.5 11.0 9.1 1.2 -2 10.7 13, 1 7.1 1.8 -3 5.6 7.1 3.8 1.9 -4 15.3 17.7 9.5 1.8 -5 5.8 5.7 6.1 0.9 - 6 6.3 6.6 5.4 1.2 Average 9.0 10.2 6.8 1.5 The percentage elongation of Ci 8 (% of Elong of Ci 8 ) in the Table 12 was calculated by dividing the sum of weight percentages for EDA, DGLA, EtrA and ETA (Table 11) by the sum of weight percentages for LA ALA, EDA, DGLA, EtrA and ETA (Table 11) and multiplying by 100 to express as a percentage. The C20 A8 desaturation percentage (C20 A8 desaturation %, Table 12) was calculated by dividing the sum of the weight percent for DGLA and ETA (Table 11) by the sum of the weight percent for EDA, DGLA, EtrA and ETA (Table 11) and when multiplying by 100 to express as a percentage. The individual elongations (% LA Elong or % ALA Elong) or A8 desaturations (% EDA A8 Desat. or % EtrA A8 Desat.) shown in Table 12 were calculated in a similar manner but only when using the co-6 substrates / products or the w-3 substrates / products for each. The elongation ratio for LA and ALA was obtained by dividing the percentage of LA elongation by the percentage of ALA elongation. Similarly, the delta-8 desaturation ratio was obtained by dividing the percentage of EDA delta-8 desaturation by the percentage of EtrA delta-8 desaturation. Example 22 Analysis of Somatic Soybean Embryos Containing EUGLENA GRACILIS Delta-8 Desaturase, ISOCHRYSIS GALBANA ELONGASE AND OTHER EPA Biosynthetic Genes (PKR786, PKR669) Plasmids pKR786 and pKR669 were digested with Ascl and DNA fragments containing the ALS selection and EPA biosynthetic genes were transformed into soybean as described above. Fatty acids from ten embryos for each strain obtained containing pKR786 and pKR669 were analyzed by GC as described. Ten embryos were analyzed for each individual transformation event. Fatty acids were identified by comparing of the retention times with those for the authentic standards. In this way, 169 events were analyzed. Of the 169 strains analyzed, 25 were identified that produced EPA (average of 10 individual embryos) at a relative abundance greater than 10% of total fatty acids. The top ten EPA producing events are shown in Table 13 and Table 14. The results for ten embryos from the best event are shown in Tables 15 and 16. The best analyzed strain had an average of 21.2% EPA with the best embryo of this lineage with 29.4% EPA (Table 16). A chromatogram for the embryo is shown in Figure 10. The fatty acids in Table 13 and Table 14 are defined as X:Y where X is the length of the fatty acid chain and Y is the number of double bonds. Furthermore, the fatty acids in Table 13 and Table 14 are further defined as follows where the number in parentheses corresponds to the position of the double bonds at the carboxyl end of the fatty acid: 18:1 = 18:1(9), 18 :2 = 18:2(9,12), GLA = 18:3(6,9,12), 18:3 = 18:3(9,12,15), STA = 18:4(6,9, 12,15), DGLA = 20:3(8,11,14), ARA = 20:4(5,8,11,14), ETA = 20:4(8,11,14,17), EPA = 20:5(5, 8,11,14,17) and DPA = 22:5(7,10,13,16,19). Fatty acids listed as "other" include: 18:2(6, 9), 20:0, 20:1(11), 20:2(8,11) and 20:3(5,11,14). Each of these fatty acids is present at a relative abundance of less than 2% of the total fatty acids. In all superior strains, GLA is either absent or present at levels below 0.2%. Table 13 Accumulation of Long Chain PUFAs in Lines Transformed with PKR786 AND PKR669 (Average of 10 Embryos per Lineage) Lineage 16:0 18:0 18:1 LA GLA ALA STA EDA DGLA ARA AFS 4314-2-1 17.0 2.6 15.6 16.8 0.1 20.4 0.3 2.7 1.2 0.1 AFS 4310-1-2 16.7 2.4 14.9 15.5 0.1 17.0 0.9 4.4 1.4 0.4 Lineage 16:0 18:0 18:1 LA GLA ALA STA EDA DGLA ARA AFS 4310-5-6 15.7 3.0 15.7 17.6 0.0 10.1 0.7 7.4 1.7 0.2 AFS 4310-1-8 15.2 2.7 16.4 15.2 0.1 17.2 0.8 4.7 1.6 0.5 AFS 4314-6-1 14.0 3, 1 12.3 17.0 0.1 8.5 0.7 11.2 2.4 0.4 AFS 4314-5-6 15.6 2.7 12.9 4.6 0.0 28.0 1 ,2 1.7 1.0 0.7 AFS 4310-7-5 17.3 1.9 9.6 16.0 0.0 22.9 0.8 2.4 2.1 1.9 AFS 4310- 1-9 14.8 2.8 13.8 12.8 0.0 17.2 0.7 4.8 1.2 0.2 AFS 4314-3-4 16.1 2.5 12.6 14, 9 0.1 18.6 0.3 3.4 1.4 0.2 AFS 4310-5-2 15.8 8.5 15.8 0.1 17.5 0.6 4.2 2.4 0 ,8 The compositions listed in Table 13 are expressed as percentage by weight of fatty acid. 16:0 = palmitic acid, 18:0 = stearic acid, 18:1 = oleic acid, LA = linoleic acid, GI_A = y-linoleic acid, AI_A = alpha-linolenic acid, STA = 5 stearidonic acid, EDA = eicosadienoic acid, DGLA = dihomo-γ-linoleic acid, ARA = arachidonic acid. Table 14 Accumulation of Long Chain PUFAs in Lines Transformed with PKR786 AND PKR669 (Average of 10 Embryos per Lineage) Lineage EtrA 20:4(5,11,14,17) ETA EPA DPA Other AFS 4314-2-1 2.4 2.1 4.0 13.6 0.1 1.0 AFS 4310-1-2 3, 6 3.6 3.1 14.1 0.4 1.6 AFS 4310-5-6 3.8 2.8 4.2 15.1 0.4 1.4 AFS 4310-1-8 2.9 2.7 3, 1 15.3 0.2 1.6 AFS 4314-6-1 4.8 3.0 4.8 15.6 0.2 1.9 AFS 4314-5-6 5.3 3.1 5.1 16.2 0.3 1.5 AFS 4310-7-5 2.4 1.6 4.0 16.6 0.1 0.5 AFS 4310-1-9 4.5 4.8 3.6 16.8 0, 3 1.8 Lineage EtrA 20:4(5,11,14,17) ETA EPA DPA Other AFS 4314-3-4 3.3 3.2 4.6 16.9 0.4 1.6 AFS 4310-5-2 3, 0 2.1 4.6 21.2 0.2 0.7 The compositions listed in Table 14 are expressed as percentage by weight of fatty acid. EtrA = eicosatrienoic acid, ETA = eicosatetraenoic acid, EPA = eicosapentaenoic acid, DPA = docosapentanoic acid. Table 15 Accumulation of Long Chain PUFAs in Lineage AFS 4310-5-2 Transformed with pKR786 and pKR669 Embryo No. 16:0 18:0 18:1 LA GLA ALA STA EDA DGLA ARA 1 16.9 2.1 8.4 20.4 0.0 18.1 0.0 2.0 1.9 0.5 2 16.3 1.8 5.3 15.6 0.1 22.3 0.4 2.2 1.9 0.9 3 17.4 3.6 10.9 14.4 0.1 21.4 0, 4 5.3 2.1 0.2 4 18.3 2.6 7.9 11.5 0.2 20.4 0.9 6.9 2.9 2.0 5 13.8 3.8 11, 1 14.5 0.1 15.0 0.7 7.7 3.0 1.0 6 15.3 3.0 11.8 15.5 0.0 18.6 0.9 4.9 1.6 0.5 7 14.3 2.0 7.6 16.4 0.2 15.9 0.6 3.2 2.1 0.3 8 15.7 2.0 5.0 17.4 0.2 12.5 0.6 3.0 3.1 1.0 9 15.1 2.2 8.1 16.3 0.2 17.0 1.1 2.6 2.8 1.3 10 15.1 1.5 8.5 15.8 0.2 13.7 0.8 4.3 2.3 0.5 Average 15.8 2.5 8.5 15.8 0.1 17.5 0.6 4 .2 2.4 0.8 The compositions listed in Table 13 are expressed as percentage by weight of fatty acid. 16:0 = palmitic acid, 18:0 = stearic acid, 18:1 = oleic acid, LA = linoleic acid, GLA = y-linoleic acid, ALA = alpha-linolenic acid, STA stearidonic acid, EDA = eicosadienoic acid, DGLA = dihomo-γ-linoleic acid, ARA = arachidonic acid. Table 16 Accumulation of Long Chain PUFAs in Lineage AFS 4310-5-2 Transformed with pKR786 and pKR669 Embryo # EtrA 20:4(5,11,14,17) ETA EPA DPA Other 1 1.9 2.1 4.5 21.1 0.0 0.0 2 2.5 2.0 5.1 22, 8 0.3 0.6 3 2.9 1.0 4.7 14.7 0.0 0.6 4 5.6 1.9 4.1 13.6 0.1 1.0 5 3.7 2 .4 3.3 18.5 0.1 1.2 6 4.1 2.5 3.6 16.4 0.1 1.1 7 2.7 2.6 5.6 25.7 0.3 0 .6 8 2.0 2.3 4.6 29.4 0.6 0.7 9 1.8 2.0 4.2 24.0 0.3 0.9 10 2.9 2.2 5.9 25.6 0.2 0.6 Average 3.0 2.1 4.6 21.2 0.2 0.7 The compositions listed in Table 14 are expressed as percentage by weight of fatty acid. EtrA = eicosatrienoic acid, ETA = eicosatetraenoic acid, EPA = eicosapentaenoic acid, DPA = docosapentaenoic acid. <110> E.I. du Pont de Nemours and Company <120> DELTA-8 DESATURASE FROM EUGLENA GRACILIS FOR USE IN THE MANUFACTURE OF POLYUNSATURATED FATTY ACIDS <130> BB1547 <150> US 60 / 583041 <151> 25-06-2004 <150> US 60 / 624812 <151> 04-11-2004 <160> 151 <170> Patentin version 3.3 <210> 1 <211 > 1271 <212> DNA <213> Euglena gracilis <220> <221> several characteristics <222> (4)..(1269) <223> Eg5: delta-8 desaturase <400> 1 gaaatgaagt caaagcgcca agcgcttccc cttacaattg atggaacaac 60 tctgcctggg 120 tcaatttcca ccctggtggt gcggaaatta tagagaatta gatgccactg 180 atgccttcat ggttatgcac tctcaagaag ccttcgacaa atgcccaaaa 240 tcaatcccag ttctgagttg ccaccccagg ctgcagtgaa gaggatttcc 300 ggaagctccg agaagagttg atcgcaactg gcatgtttga ctctggtact 360 catacaaaat cagcaccaca ctgggccttg gagtgctggg atggttcagt 420 atcagatgta tttcattggg gcagtgttgc ttgggatgca atgggctggc 480 tttctcatga catttgccac caccagactt tcaagaaccg aacctcgtgg 540 gactggtatt tggcaatggt ctgcaaggtt tttccgtgac gacagacaca 600 atgcacatca ttcggcaacc aatgttcaag ggcacgaccc aacctccccc 660 tcttagcctg gtctgaggat gacgtcacac gggcgtcacc aagctcattc 720 agttccagca gtactatttc ttggtcatct gtatcttgtt tggtgtttcc 780 agagcgtgtt gaccgtgcgc agtttgaagg acagagataa cgctctcagt 840 ataagaagga ggccattggc ctcgccctgc actggacctt ttccacttat 900 tctttatgcc cagcatcctc acatcgctgt tggtgttttt atatgatgtg ccaaggaagg gctcaagcgc tgaagctcaa tgccctccccc ttatttcctg ctatcaacag gaactggaac atggtggaag tgatattgac gatttcccgc gcggttcatt ccaattctat gaagaccctg cgtttcggag ctggttggcg 960 atcggggact 1020 aacattcggc 1080 caccatttgt 1140 cagctgtgcc 1200 atcctgctgc 1260 gctctataag 1271 <210> 2 <211> 421 <212> PRT <213> Eug. gcttcggcat cagtctggga gagggattat ggccgaccct agaagcacaa gctatctggc g wow grace. tgcgatcgtg tggccatgga cacagattgg ccctcgccac cctgccgtat ggtgttcgcc is gtgttcatga ttctcggttg tttttcggag aacctgacag cggaacccgc cggatggcgg accactaccc gccagatcca gcttgaatta cggttagcta tgccccatga agaagcaacc actggagaag tgagaccatg ccagattgag ccaggtggaa agggttggtc cgcggggaag <400> 2 Met 1 Lys Ser Lys Arg 5 Gin Ala Leu Pro Leu 10 Thr He Asp Gly Thr 15 Thr Tyr Asp Vai Ser 20 Ala Trp Vai Asn Phe 25 His Pro Gly Gly Ala 30 Glu He He Glu Asn 35 Tyr Gin Gly Arg Asp 40 Ala Thr Asp Ala Phe 45 Met Vai Met His Ser 50 Gin Glu Ala Phe Asp 55 Lys Leu Lys Arg Met 60 Pro Lys He Asn Pro 65 Ser Ser Glu Leu Pro 70 Pro Gin Ala Ala Vai 75 Asn Glu Ala Gin Glu 80 Asp Phe Arg Lys Leu 85 Arg Glu Glu Leu He 90 Ala Thr Gly Met Phe 95 Asp Ala Ser Pro Leu 100 Trp Tyr Ser Tyr Lys 105 He Ser Thr Thr Leu 110 Gly Leu Gly Vai Leu 115 Gly Tyr Phe Leu Met 120 Vai Gin Tyr Gin Met 125 Tyr Phe He Gly Ala 130 Vai Leu Leu Gly Met 135 His Tyr Gin Gin Met 140 Gly Trp Leu Ser His 145 Asp He Cys His His 150 Gin Thr Phe Lys Asn 155 Arg Asn Trp Asn Asn 160 Leu Vai Gly Leu Vai 165 Phe Gly Asn Gly Leu 170 Gin Gly Phe Ser Vai 175 Thr Trp Trp Lys Asp Arg His Asn Ala His His Ser Ala Thr Asn Vai Gin 180 185 190 Gly His Asp Pro Asp lie Asp Asn 195 200 Asp Asp Vai Thr Arg Ala Ser Pro 210 215 Gin Gin Tyr Tyr Phe Leu Vai lie 225 230 Cys Phe Gin Ser Vai Leu Thr Vai 245 Gin Phe Tyr Arg Ser Gin Tyr Lys 260 His Trp Thr Leu Lys Thr Leu Phe 275 280 Leu Thr Ser Leu Leu Vai Phe Phe 290 295 Glylie Ala He Vai Vai Phe Met 305 310 Gly Asp Ser Vai Trp Asp Gly His 325 Glu Thr Met Asn He Arg Arg Gly 340 Gly Leu Asn Tyr Gin lie Glu His 355 360 His Asn Leu Thr Ala Will Be Tyr 370 375 His Asn Leu Pro Tyr Arg Asn Pro 385 390 Leu Leu Arg Tyr Leu Ala Vai Phe 405 Ala Gly Lys Ala Leu <210> 3 <211> 1271 <212> DNA <213> Euglena gracilis <220> <221> various features <222> (4)..(1269) <223> Egl2: delta-8 desaturase <400> 3 gaaatgaagt caaagcgcca agcgcttccc 60 Leu Pro Leu Leu Ala Trp Ser Glu 205 Lie Ser Arg Lys Leu Lie Gin Phe 220 Cys lie Leu Leu Arg Phe lie Trp 235 240 Arg Ser Leu Lys Asp Arg Asp Asn 250 255 Lys Glu Ala He Gly Leu Ala Leu 265 270 His Leu Phe Phe Met Pro Ser lie 285 Vai Ser Glu Leu Vai Gly Gly Phe 300 Asn His Tyr Pro Leu Glu Lys lie 315 320 Gly Phe Ser Vai Gly Gin lie His 330 335 He He Thr Asp Trp Phe Phe Gly 345 350 His Leu Trp Pro Thr Leu Pro Arg 365 Gin Vai Glu Gin Leu Cys Gin Lys 380 Leu Pro His Glu Gly Leu Vai He 395 400 Ala Arg Met Ala Glu Lys Gin Pro 410 415 cttacaattg atggaacaac atatgatgtg tctgcctggg 120 gatgccactg 180 atgcccaaaa 240 gaggatttcc 300 ctctggtact 360 atggttcagt 420 atgggctggc 480 aacctcgtgg 540 gacagacaca 600 aacctccccc 660 aagctcattc 720 tggtgtttcc 780 cgctctcagt 840 ttccacttat 900 ctggttggcg 960 atcggggact 1020 aacattcggc 1080 caccatttgt 1140 cagctgtgcc 1200 atcctgctgc 1260 gctctataag 1271 tcaatttcca atgccttcat tcaatcccag ggaagctccg catacaaaat atcagatgta tttctcatga gactggtatt atgcacatca tcttagcctg agttccagca agagcgtgtt ataagaagga tctttatgcc gcttcggcat cagtctggga gagggattat ggccgaccct agaagcacaa gctatctggc g ' ccctggtggt ggttatgcac ttctgagttg agaagagttg cagcaccaca tttcattggg catttgccac tggcaatggt ttcggcaacc gtctgaggat gtactatttc gaccgtgcgc ggccattggc cagcatcctc tgcgatcgtg tggccatgga cacagattgg ccctcgtccat cctgccg gcggaaatta tctcaagaag ccaccccagg atcgcaactg ctgggccttg gcagtgttgc caccagactt ctgcaaggtt aatgttcaag gacgtcacac ttggtcatct agtttgaagg ctcgccctgc acatcgctgt gtgttcatga ttctcggggggttg tttttcgg aacctatgaccgc cgg tagagaatta ccttcgacaa ctgcagtgaa gcatgtttga gagtgctggg ttgggatgca tcaagaaccg tttccgtgac ggcacgaccc gggcgtcacc gtatcttgtt acagagataa actggacctt tggtgttttt accactaccc gccagatcca gcttgaatta cggttagcta tgccaccatga agagataa ccaaggaagg gctcaagcgc tgaagctcaa tgcctcccctg ctatcaacag gaactggaac atggtggaag tgatattgac gatttcccgc gcggttcatt ccaattctat gaaggccctg cgtttcggag actggagaag tgagaccatg ccagattgaggg ccaggtggaa agggtctggagg c <210> 4 <211> 421 <212> PRT <213> Euglena gracilis <400> 4 Met Lys Ser Lys 1 Tyr Asp Will Be Arg Gin Ala Leu Pro Ala Trp Vai Asn Phe Read thr lie His Pro Gly Asp Gly Thr Thr Gly Ala Glulie He Glu Asn Tyr Gin Gly Arg Asp Ala Thr Asp Ala Phe Met Vai Met 35 40 45 His Ser 50 Gin Glu Ala Phe Asp Lys 55 Leu Lys Arg Met 60 Pro Lys lie Asn Pro Ser Ser Glu Leu Pro Gin Ala Ala Vai Asn Glu Ala Gin Glu 65 70 75 80 Asp Phe Arg Lys Leu Arg Glu Glu Leu He Ala Thr Gly Met Phe Asp 85 90 95 Ala Ser Pro Leu Trp Tyr Ser Tyr Lys He Ser Thr Thr Leu Gly Leu 100 105 110 Gly Vai Leu Gly Tyr Phe Leu Met Vai Gin Tyr Gin Met Tyr Phe lie 115 120 125 Gly Ala Vai Leu Leu Gly Met His Tyr Gin Gin Met Gly Trp Leu Ser 130 135 140 His Asp He Cys His His Gin Thr Phe Lys Asn Arg Asn Trp Asn Asn 145 150 155 160 Leu Vai Gly Leu Vai Phe Gly Asn Gly Leu Gin Gly Phe Ser Vai Thr 165 170 175 Trp Trp Lys Asp Arg His Asn Ala His His Ser Ala Thr Asn Vai Gin 180 185 190 Gly His Asp Pro Asp He Asp Asn Leu Pro Leu Leu Ala Trp Ser Glu 195 200 205 Asp Asp Vai Thr Arg Ala Ser Pro He Ser Arg Lys Leu He Gin Phe 210 215 220 Gin Gin Tyr Tyr Phe Leu Vai He Cys He Leu Leu Arg Phe He Trp 225 230 235 240 Cys Phe Gin Ser Vai Leu Thr Vai Arg Ser Leu Lys Asp Arg Asp Asn 245 250 255 Gin Phe Tyr Arg Ser Gin Tyr Lys Lys Glu Ala He Gly Leu Ala Leu 260 265 270 His Trp Thr Leu Lys Ala Leu Phe His Leu Phe Phe Met Pro Ser He 275 280 285 Leu Thr Ser Leu Vai Phe Phe Vai Ser Glu Leu Vai Gly Gly Phe 290 295 300 Gly He Ala He Vai Phe Met Asn His Tyr Pro Leu Glu Lys He 305 310 315 320 Gly Asp Ser Vai Trp Asp Gly His Gly Phe Ser Vai Gly Gin He His 325 330 335 Glu Thr Met Asn He Arg Arg Gly He He Thr Asp Trp Phe Phe Gly 340 345 350 Gly Leu Asn Tyr Gin He Glu His His Leu Trp Pro Thr Leu Pro Arg 355 360 365 His Asn 370 Leu Thr Ala Vai Ser 375 Tyr Gin Vai Glu Gin 380 Leu Cys Gin Lys His Asn Leu Pro Tyr Arg Asn Pro Leu Pro His Glu Gly Leu Vai lie 385 390 395 400 Leu Leu Arg Tyr Leu Ala Vai Phe Ala Arg Met Ala Glu Lys Gin Pro 405 410 415 Ala Gly Lys Ala Leu <210> 5 <211> 1275 <212> DNA <213> Euglena gracilis (GenBank Accession Nos. AF139720 and AAD45877) <220> <221> several features <222> (14)..(1273) <223> non-functional delta 8 desaturase <400> 5 attttttttc 60 gaaatgaagt caaagcgcca agcgctatcc cccttacaat tgatggaaca aacatatgat 120 gtggtcaatt tccaccctgg tggtgcggaa attatagaga attaccaagg aagggatgcc 180 actgatgcct tcatggttat gcactttcaa gaagccttcg acaagctcaa gcgcatgccc 240 aaaatcaatc ccagttttga gttgccaccc caggctgcag tgaatgaagc tcaagaggat 300 ttccggaagc tccgagaaga gttgatcgca actggcatgt ttgatgcctc ccccctctgg 360 tactcataca aaatcagcac cacactgggc cttggagtgc tgggttattt cctgatggtt 420 cagtatcaga tgtatttcat tggggcagtg ttgcttggga tgcactatca acagatgggc 480 tggctttctc atgacatttg ccaccaccag actttcaaga accggaactg gaacaacctc 540 gtgggactgg tatttggcaa tggtctgcaa ggtttttccg tgacatgttg gaaggacaga 600 cacaatgcac atcattcggc aaccaatgtt caagggcacg accctgatat tgacaacctc 660 ccccccttag cctggtctga ggatgacgtc acacgggcgt caccgatttc ccgcaagctc 720 attcagttcc agcagtacta tttcttggtc atctgtatct tgttgcggtt catttggtgt 780 ttccagtgcg tgttgaccgt gcgcagtttg aaggacagag ataaccaatt ctatcgctct 840 cagtataaga aggaggccat tggcctcgcc ctgcactgga ccttgaaggc cctgttccac 900 ttattc ttta tgcccagcat cctcacatcg ctgttggtgt ttttcgtttc ggagctggtt 960 ggcggcttcg gcattgcgat cgtggtgttc atgaaccact acccactgga gaagatcggg 1020 gacccagtct gggatggcca tggattctcg gttggccaga tccatgagac catgaacatt cggcgaggga ttatcacaga ttggtttttc ggaggcttga attaccagat 1080 tgagcaccat ttgtggccga ccctccctcg ccacaacctg acagcggtta gctaccaggt 1140 ggaacagctg tgccagaagc acaacctgcc gtatcggaac ccgctgcccc atgaagggtt 1200 ggtcatcctg ctgcgctatc tggcggtgtt cgcccggatg gcggagaagc aacccgcggg 1260 gaaggctcta taagg <210> 6 <211> 419 <212> PRT <213> Euglena gracilis (GenBank Accession No. AF139720 and AAD45877) <400> 6 Met 1 Lys Ser Lys Arg Gin Ala 5 Leu Ser Pro 10 Leu Gin Leu Met Glu 15 Gin Thr Tyr Asp Vai Vai Asn Phe His Pro Gly Gly Ala Glu He He Glu 20 25 30 Asn Tyr Gin Gly Arg Asp Ala Thr Asp Ala Phe Met Vai Met His Phe 35 40 45 Gin Glu Ala Phe Asp Lys Leu Lys Arg Met Pro Lys He Asn Pro Ser 50 55 60 Phe Glu Leu Pro Pro Gin Ala Ala Vai Asn Glu Ala Gin Glu Asp Phe 65 70 75 80 Arg Lys Leu Arg Glu Glu Leu He Ala Thr Gly Met Phe Asp Ala Ser 85 90 95 Pro Leu Trp Tyr Ser Tyr Lys He Ser Thr Thr Leu Gly Leu Gly Vai 100 105 110 Leu Gly Tyr Phe Leu Met Vai Gin Tyr Gin Met Tyr Phe He Gly Ala 115 120 125 Vai Leu Leu Gly Met His Tyr Gin Gin Met Gly Trp Leu Ser His Asp 130 135 140 lie Cys His Gin Thr Phe Lys Asn Arg Asn Trp Asn Asn Leu Vai 145 150 155 160 Gly Leu Vai Phe Gly Asn Gly Leu Gin Gly Phe Ser Vai Thr Cys Trp 165 170 175 Lys Asp Arg His Asn Ala His His Ser Ala Thr Asn Vai Gin Gly His 180 185 190 Asp Pro Asp lie Asp Asn Leu Pro Pro Leu Ala Trp Ser Glu Asp Asp 195 200 205 Vai Thr Arg Ala Ser Pro He S er Arg Lys Leu lie Gin Phe Gin Gin 210 215 220 Tyr Tyr Phe Leu Vai lie Cys He Leu Leu Arg Phe He Trp Cys Phe 225 230 235 240 Gin Cys Vai Leu Thr 245 Vai Arg Ser Leu Lys Asp 250 Arg Asp Asn Gin 255 Phe Tyr Arg Ser Gin Tyr Lys Lys Glu Ala He Gly Leu Ala Leu His Trp 260 265 270 Thr Leu Lys Ala Leu Phe His Leu Phe Phe Met Pro Ser He Leu Thr 275 280 285 Ser Leu Leu Vai Phe Phe Vai Ser Glu Leu Vai Gly Gly Phe Gly He 290 295 300 Ala He Vai Vai Phe Met Asn His Tyr Pro Leu Glu Lys lie Gly Asp 305 310 315 320 Pro Vai Trp Asp Gly His Gly Phe Ser Vai Gly Gin lie His Glu Thr 325 330 335 Met Asn He Arg Arg Gly He He Thr Asp Trp Phe Gly Gly Gly Leu 340 345 350 Asn Tyr Gin lie Glu His His Leu Trp Pro Thr Leu Pro Arg His Asn 355 360 365 Leu Thr Ala Vai Ser Tyr Gin Vai Glu Gin Leu Cys Gin Lys His Asn 370 375 380 Leu Pro Tyr Arg Asn Pro Leu Pro His Glu Gly Leu Vai He Leu Leu 385 390 395 400 Arg Tyr Leu Ala Vai Phe Ala Arg Met Ala Glu Lys Gin Pro Ala Gly 405 410 415 Lys Wing Leu <210> 7 <211> 422 <212> PRT <213> Euglena gracilis <400> 7 Met 1 Lys Ser Lys Arg 5 Gin Ala Leu Ser Pro 10 Leu Gin Leu Met Glu 15 Gin Thr Tyr Asp Vai 20 Ser Ala Trp Vai Asn 25 Phe His Pro Gly Gly 30 Ala Glu Helie Glu 35 Asn Tyr Gin Gly Arg 40 Asp Ala Thr Asp Ala 45 Phe Met Vai Met His 50 Phe Gin Glu Ala Phe 55 Asp Lys Leu Lys Arg 60 Met Pro Lys He Asn 65 Pro Ser Phe Glu Leu 70 Pro Pro Gin Ala Ala 75 Vai Asn Glu Ala Gin 80 Glu Asp Phe Arg Lys Leu Arg Glu Glu Leu Ile Ala Thr Gly Met Phe 85 90 95 Asp Ala Ser Pro Leu Trp Tyr Ser Tyr Lys Ile Ser Thr Thr Leu Gly 100 105 110 Leu Gly Vai Leu Gly Tyr Phe Leu Met Vai Gin Tyr Gin Met Tyr Phe 115 120 125 lie Gly Ala Vai Leu Gly Met His Tyr Gin Gin Met Gly Trp Leu 130 135 140 Ser His Asp Ile Cys His Gin Thr Phe Lys Asn Arg Asn Trp Asn 145 150 155 160 Asn Leu Vai Gly Leu Vai Phe Gly Asn Gly Leu Gin Gly Phe Ser Vai 165 170 175 Thr Cys Trp Lys Asp Arg His Asn Ala His His Ser Ala Thr Asn Vai 180 185 190 Gin Gly His Asp Pro Asp Ile Asp Asn Leu Pro Leu Ala Trp Ser 195 200 205 Glu Asp Asp Vai Thr Arg Ala Ser Pro Ile Ser Arg Lys Leu Ile Gin 210 215 220 Phe Gin Gin Tyr Phe Leu Vai lie Cys Ile Leu Arg Phe Ile 225 230 235 240 Trp Cys Phe Gin Cys Vai Leu Thr Vai Arg Ser Leu Lys Asp Arg Asp 245 250 255 Asn Gin Phe Tyr Arg Ser Gin Tyr Lys Lys Glu Ala Ile Gly Leu Ala 260 265 270 read his Trp Thr Leu Lys Ala Leu Phe His Leu Phe Phe Met Pro Ser 275 280 285 Ile Leu Thr Ser Leu Leu Vai Phe Phe Vai Ser Glu Leu Vai Gly Gly 290 295 300 Phe Gly Ile Ala Ile Vai Vai Phe Met Asn His Tyr Pro Leu Glu Lys 305 310 315 320 Ile Gly Asp Pro Vai Trp Asp Gly His Gly Phe Ser Vai Gly Gin Ile 325 330 335 His Glu Thr Met Asn Ile Arg Arg Gly Ile Ile Thr Asp Trp Phe Phe 340 345 350 Gly Gly Leu Asn Tyr Gin Ile Glu His His Leu Trp Pro Thr Leu Pro 355 360 365 Arg His Asn Leu Thr Ala Vai Ser Tyr Gin Vai Glu Gin Leu Cys Gin 370 375 380 Lys His Asn Leu Pro Tyr Arg Asn Pro Leu Pro His Glu Gly Leu Vai 385 390 395 400 He Leu Leu Arg Tyr Leu Ala Vai Phe Ala Arg Met Ala Glu Lys Gin 405 410 415 Pro Ala Gly Lys Ala Leu <210> <211> <212> <213> 8 19 DNA Artificial Sequence <220> <223> Primer Eg5-1 <400>8 gaaatgaagt caaagcgcc 19 <210> <211> <212> <213> 9 19 DNA Artificial Sequence 220> <223> Primer Eg3-3 <400> 9 ccttatagag cctccccg 19 <210> <211> <212> <213> 10 22 DNA Artificial Sequence <220 <223> Primer T7 <400> 10 gtaatacgac tcactatagg gc 22 <210> <211> <212> <213> 11 19 DNA Artificial Sequence <220> <223> Primer M13-28Rev <400> 11 ggaaacagct atgaccatg <210> 12 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Primer Eg3-2 <400> 12 ttggcaatgg tctgcaagg 19 <210> 13 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Primer Eg5-2 <400> 13 aatgttcatg gtctcatgg <210> 14 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> <400> 14 ggatctcctg caggatctgg ccggccggat ctcgtac 37 <210> 15 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Primer RPB2forward <400> 15 gcggccgcat ggagtcgatt gcgc 24 <210> 16 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Primer RPB2reverse <400> 16 gcggccgctt actgcaactt cctt <210> 17 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> <400> 17 aagcttgcat gcctgcaggt cgactcgacg tacg <210> 18 <211> 280 <212> DNA <213> soy albumin transcriptional terminator <400> 18 tctagaggat ccaaggccgc gaagttaaaa gcaatgttgt cacttgtcgt 60 gatgtgatag tttatgctag ctagctataa cataagctgt ctctgagtgt 120 aataaagatc atcactggtg aatggtgatc gtgtacgtac cctacttagt 180 agcacttaga gtgtgctttg tgcatggcct tgccctctgtt ttgagacttt 240 tcgagtttaa atctttgcct ttgcgtacgt gggcggatcc actaacacat gttgtatatt aggcaatgga tgtaatgttt <210> 19 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> Primer oSalb-12 <400> 19 tttggatcct ctagacgtac gcaaaggcaa ag <210> 20 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> Primer oSalb-13 <400> 20 aaaggatcca aggccgcgaa gttaaaagca atgttg <210> 21 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Primer GSP1 <400> 21 gccccccatc ctttgaaagc ctgt <210> 22 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> Primer GSP2 <400> 22 cgcggatccg agagcctcag catcttgagc agaa <210> 23 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Primer GSP3 <400> 23 ggtccaatat ggacgatga gttgata <210> 24 <211> 35 <212> DNA <213> Artificial Sequence <220> <223> Primer GSP4 <400> 24 cgcggatccg ctggaactag aagagagacc taaga <210> 25 <211> 1408 <212> DNA <213> Glycine max <220> <221> several characteristics <223> BD30 promoter <400> 25 aactaaaaaa agctctcaaa ttacattttg agttgtttca ggttccattg aaaactccaa ctaaaataac aaatagcaca tgeaggtgea aacaacacgt 120 aaggtgatgt gcctctagca gtetagetta tgaggetege tgcttatcaa ccttattgct tactctgatg cgattcatca ttccccaaga cgtgtacgca gattaaacaa tggacaaaac ttcaatcgat tatagaataa 240 taattttaac agtgccgact tttttctgta aacaaaaggc cagaatcata tcgcacatca 300 tcttgaatgc agtgtcgagt ttggaccatt tgagtacaaa gccaatattg aatgattttt 360 cgattttaca tgtgtgaatc agacaaaagt gcatgcaatc acttgcaagt aaattaagga 420 tactaatcta ttcctttcat tttatatgct ccacttttat ataaaaaaat atacattatt 480 atatatgcat tattaattat tgcagtatta tgctattggt tttatggccc tgctaaataa 540 cctaaatgag tctaactatt gcatatgaat caaatgaagg aagaatcatg atctaaacct 600 gagtacccaa tgcaataaaa tgcgtcctat tacctaaact tcaaacacac attgccatcg 660 gacgtataaa ttaatgcata taggttattt tgagaaaaga aaacatcaaa agctctaaaa 720 cttcttttaa ctttgaaata agctgataaa aatacgcttt aaatcaactg tgtgctgtat 780 ataagctgca atttcacatt ttaccaaacc gaaacaagaa tggtaacagt gaggcaaaaa 840 tttgaaaaat gtcctacttc acattcacat caaattaatt acaactaaat aaataaacat 900 cgtgattcaa gcagtaatga aagtcgaaat cagatagaat atacacgttt aacatcaatt 960 gaattttttt ttaaatggat atatacaagt ttactatttt atatataatg aaaattcatt 1020 ttgtgttagc acaaaact ta cagaaagaga taaattttaa ataaagagaa ttatatccaa 1080 ttttataatc caaaataatc aaattaaaga atattggcta gatagaccgg ctttttcact 1140 gcccctgctg gataatgaaa attcatatca aaacaataca gaagttctag tttaataata 1200 aaaaagttgg caaactgtca ttccctgttg gtttttaagc caaatcacaa ttcaattacg 1260 tatcagaaat taatttaaac caaatatata gctacgaggg aacttcttca gtcattacta 1320 gctagctcac taatcactat atatacgaca tgctacaagt gaagtgacca tatcttaatt 1380 tcaaatcata aaattcttcc accaagtt 1408 <210> 26 <211> 690 <212> DNA <213> Glycine i max <220> <221> Miscellaneous characteristics <223> Glycinin Gyl Promoter <400> 26 tagcctaagt acgtactcaa aatgccaaca aataaaaaaa aagttgcttt aataatgcca 60 aaacaaatta ataaaacact tacaacaccg gatttttttt aattaaaatg tgccatttag 120 gataaatagt taatattttt aataattatt taaaaagccg tatctactaa aatgattttt atttggttga aaatattaat atgtttaaat caacacaatc tatcaaaatt aaactaaaaa 240 aaaaataagt gtacgtggtt aacattagta cagtaatata agaggaaaat gagaaattaa 300 gaaattgaaa gcgagtctaa tttttaaatt atgaacctgc atatataaaa ggaaagaaag 360 aatccaggaa gaaaagaaat gaaaccatgc atggtcccct cgtcatcacg agtttctgcc 420 atttgcaata gaaacactga aacacctttc tctttgtcac ttaattgaga tgccgaagcc 480 acctcacacc atgaacttca tgaggtgtag cacccaaggc ttccatagcc atgcatactg 540 aagaatgtct caagctcagc accctacttc tgtgacgttg tccctcattc accttcctct 600 cttccctata aataaccacg cctcaggttc tccgcttcac aactcaaaca ttctccctcca 660 ttggtcctta aacactcatc agtcatcacc <210> 27 <211> 36 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 27 cgcggatcct agcctaagta cgtactcaaa atgcca <210> 28 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 28 gaattcgcgg ccgcggtgat gactgatgag tgtttaagga c <210> 29 <211> 2012 <212> DNA <213> Glycine max <220> <221> various features <223> annexin promoter <400> 29 atcttaggcc cttgattata tggtgtttag atggattcac atgcaagttt ttatttcaat 60 cccttttcct ttgaataact gaccaagaac aacaagaaaa aaaaaaaaag aaaaggatca 120 ttttgaaagg atatttttcg ctcctattca aatactgtat ttttaccaaa aaactgtat ctctcaagct tttataaatg gtacaaatta acatctaagt gactattttg gaaatattgt ttattatatt atgtaaagta ctgatatgaa ttttctttca atttataaaa ttatgattga tttcctaaaa catttttctc tactctcttt attggggcat aaagcattgg ttgattcagt atcttaaata ttttgtttag taaacatata taaggataaa aatattttag ctatatgtat tcttgccatt ctcttcacac tttcttttta gtgccaaaag gaaaaagcat ttttcctaca 240 aatcactcta 300 agcttgcaat 360 tattaatcta 420 atttccccgt 480 tacatgaaaa 540 atatagttaa 600 atttatgcat 660 taaaaaaaga 720 ttttcttttc 780 ttatttatca 840 aatcatatca 900 aaatgtgtta 960 tctctcattt 1020 taattatttt 1080 acactccaaa 1140 atgaaactga 1200 tggcatctga 1260 ctattaatta 1320 atatgttatt 1380 ttatattgat 1440 cataatttta 1500 gagaaaaaaa 1560 tcataaattc 1620 ctacgtcatc 1680 ccatgaatca 1740 tctctttccg 1800 atttattaac 1860 atttagaacat 1920 cttcgactct gtgtgaacaa tgcctttcaa aattttttcg caacactggc aattaaaaat ctattctatt ttgctatata tttttaaagc aatttaataa ttatatgttt attgacagtg caaacatgta tctttatttt tctatccctc ctactctatc cccctcctca catgtgtaaa aaaatatttt gaataaatct aatatgatat aaaataaatt atgaccagca aaaaacagat gcaaacagaa agcaaacaag tttgaaatag aatagagtgc cgataatcct ttgtttttcg gcataaggta atgtttttca aaggctagtg gctttgactc ttatattctt acttttttct cattatattt aaatttattc ttttctattc atattttact aagagaggaa aaaacctaaa tttatctctt attttttctc gcctttatca caagtctcct atatctattg aataaaggat atgtatattc atcaatatta aaatatgata ccataaataa atttgaaatt tgtttttcat catttttact caacactttc atgcttattg tgttctaaca catgatttcc ttgcaaagct aattatctgc aatagtcatg cccgtacatc attgtccctt agttctattt cttaaatatt tatatcattt gaaataaatt tttcacattt agtaataagt tctactctct tttatcatat tcttatttcc tcctccacta agtgatttcc agtctttcca tttttcttca aaatatatga ttttttatat ttaaatattt tattttacaa aatcgttaag agggacgaaa tctcaacaac tagtattttt acatttgtaa gtgaaggcgt ttcatatgcc tgtcatcaaa accccctagc cagtgcatta cgtccctcat ctaaattttc tcaaattatt tctaaattat tatatatact ttgtaaattt ttgttaaaca gatgagaaaa atttcatcta catttcacat actcatatat aatcatttaa aactcagcat caatacataa taaaattaaa taatgatttt aggttgtaca ttatatttac ccttagtaat ggaaggaatc aaccacctca tgaagccagc ttttcacatg tgtactacta taactccaaa gatccaattt cactatataa attgtgacga aagcaaaatg aattcacata gctgagagag 1980 aaaggaaagg ttaactaaga agcaatactt 2012 ca <210> 30 <211> 37 <212> DNA <213> Seqüência Artificial <220> <223> Primer <400> 30 cgcggatcca tcttaggccc ttgattatat ggtgttt 37 <210> 31 < 211> 43 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 31 gaattcgcgg ccgctgaagt attgcttctt agttaacctt tcc 43 <210> 32 <211> 41 <212> DNA Artificial Sequence <220> <223> Primer <400> 32 cgcggatcca actaaaaaaa gctctcaaat tacattttga g 41 <210> 33 <211> 44 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 33 gaattcgcgg ccgcaacttg gtggaagaat tttatgattt gaaa <210> 34 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> Primer oKTis <400> 34 atctagacgt acgtcctcga agagaaggg 29 <210> <211> <212> <213> 35 22 DNA Artificial Sequence <220> <223> Primer 0KTÍ6 <400> 35 ttctagacgt accgatataa tg <210> <211> <212> <213> 36 17 DNA Artificial Sequence <220> <223> Primer OSBD30-1 <400> 36 tgcggccgca tgagccg 17 <210> <211> <212> <213> 37 32 DNA Artificial Sequence <220> <223> Primer OSBD30-2 <400> 37 acgtacggta ccatctgcta atattttaaa tc 32 <210> <211> <212> <213> 38 32 DNA Artificial Sequence <220> <223> Primer OCGR5-1 <400> 38 ttgcggccgc aaaccatggc tgctgctccc ag 32 <210> <211> <212> <213> 39 24 DNA Artificial Sequence <223> Primer oCGR5-2 <400> 39 aagcggccgc ttactgcgcc ttac 24 <210> 40 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> Primer oSGly-1 <400> 40 ttcctgcagg ctagcctaag tacgtactc <210> 41 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer oSGly-2 <400> 41 aagcggccgc ggtgatgact g <210> 42 <211> 36 <212> DNA <213> Artificial Sequence <220> <223 > Primer LegPro5' <400> 42 tttctagacg tacgtccctt cttatctttg atctcc <210> 43 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> Primer LegPro3 1 <400> 43 gcggccgcag ttggatagaa tatatgtttg tgac <210> 44 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> Primer LegTermS' <400> 44 ctatccaact gcggccgcat ttcgcaccaa atcaatgaaa g <210> <211> <212> <213> 45 38 DNA Artificial Sequence <220> <223> Primer LegTerm3' <400> 45 aatctagacg tacgtgaagg ttaaacatgg tgaatatg 38 <210> <211> <212> <213> 46 24 DNA Artificial Sequence <220> <223> Primer CGR4forward <400> 46 gcggccgcat gggaacggac caag 24 <210> <211> <212> <213> 47 24 DNA Artificial Sequence <220> <223> Primer CGR4reverse <400> 47 gcggccgcct actcttcctt ggga 24 <210> <211> <212> <213> 48 1270 DNA Artificial Sequence <220> <223> D8S-1: Codon-optimized synthetic gene for expression in Yarrowia lipolytica <400> 48 ccatggagtc caagcgacag gctctgtctc ccctccagct gatggaacag acctacgacg 60 tcgtgaactt ccaccctggt ggagctgaaa tcattgagaa ctaccaggga cgagatgcta ctgacgcctt catggttatg cactttcagg aagccttcga caagctcaag cgaatgccca agatcaaccc ctcctttgag ctgcctcccc aggctgccgt caacgaagct caggaggatt tccgaaagct ccgagaagag ctgatcgcca ctggcatgtt tgacgcctct cccctctggt actcgtacaa gatctccacc accctgggtc ttggcgtgct tggatacttc ctgatggtcc 360 agtaccagat gtacttcatt ggtgctgtgc tgctcggtat gcactaccag caaatgggat ggctgtctca tgacatctgc caccaccaga ccttcaagaa ccgaaactgg aataacctcg 480 tgggtctggt ctttggcaac ggactccagg gcttctccgt gacctgttgg aaggacagac acaacgccca tcattctgct accaacgttc agggtcacga tcccgacatt gataacctgc ctcccctcgc ctggtccgag gacgatgtca ctcgagcttc tcccatctcc cgaaagctca ttcagttcca acagtactat ttcctggtca tctgtattct cctgcgattc atctggtgtt tccagtgcgt gctgaccgtt cgatccctca aggaccgaga caaccagttc taccgatctc agtacaagaa agaggccatt ggactcgctc tgcactggac tctcaaggct ctgttccacc tcttctttat gccctccatc ctgacctcgc tcctggtgtt ctttgtttcc gagctcgtcg 900 gtggcttcgg aattgccatc gtggtcttca tgaaccacta ccctctggag aagatcggtg 960 atcccgtctg ggacggacat ggcttctctg tgggtcagat ccatgagacc atgaacattc 1020 gacgaggcat cattactgac tggttctttg gaggcctgaa ctaccagatc gagcaccatc 1080 tctggcccac cctgcctcga cacaacctca ctgccgtttc ctaccaggtg gaacagctgt 1140 gccagaagca caacctcccc taccgaaacc ctctgcccca tgaaggtctc gtcatcctgc 1200 tccgatacct ggccgtgttc gctcgaatgg cccgagaagca gcccgctggc aaggctctct 1260 aagcggccgc <210> 49 <211> 104 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-1A <400> 49 atggagtcca agcgacaggc tctgtctccc ctccagctga tggaacagac ctacgacgtc 60 gtgaacttcc accctggtgg agctgaaatc attgagaact acca <210> 50 <211> 104 <212> DNA <213> Artificial Sequence <223> Primer D8-1B <400> 50 tccctggtag ttctcaatga tttcagctcc accagggtgg aagttcacga cgtcgtaggt 60 ctgttccatc agctggaggg gagacagagc ctgtcgcttg gact 104 <210> 51 <211> 102 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-2A <400> 51 gggacgagat gctactgacg ccttcatggt tatgcacttt caggaagcct tcgacaagct caagcgaatg cccaagatca accccctcctt tgagctgcct cc <210> 52 <211> 102 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-2B <400> 52 ctggggaggc agctcaaagg aggggttgat cttgggcatt cgcttgagct tgtcgaaggc ttcctgaaag tgcataacca tgaaggcgtc agtagcatct eg <210> 53 <211> 101 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-3A <400> 53 ccaggctgcc gteaaegaag ctcaggagga tttccgaaag ctccgagaag agetgatege cactggcatg tttgacgcct ctcccctctg gtactcgtac a <210> 54 <211> 101 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-3B <400> 54 atcttgtacg agtaccagag ggggagaggcg tcaaacatgc cagtggcgat cagctcttct 60 cggagctttc ggaaatcctc ctgagcttcg ttgacggcag c 101 <210> 55 <211> 101 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-4A <400> 55 ccaccaccct gggtcttggc gtgcttggat 60 tcattggtgc tgtgctgctc ggtatgcact 101 acttcctgat accagcaaat ggtccagtac g cagatgtact <210> 56 <211> 101 <212> DNA <213> Seqüência Artificial <220> <223> Primer D8-4B <400> 56 atcccatttg ctggtagtgc ataccgagca 60 gcacagcacc aatgaagtac atctggtact ggaccatcag gaagtatcca agcacgccaa 101 gacccagggt g <210> 57 <211> 104 <212> DNA <213> Seqüência Artificial <220> <223> Primer D8-5A <400> 57 ggatggctgt ctcatgacat ctgccaccac 60 cagaccttca agaaccgaaa ctggaataac ctcgtgggtc tggtctttgg caacggactc cagggcttct ccgt <210> 58 <211> 104 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-5B <400> 58 ggtcacggag aagccctgga gtccgttgcc aaagaccaga cccacgaggt tattccagtt 60 tcggttcttg aaggtctggt ggtggcagat gtcatgagac agcc <210> <211> <212> <213> 59 101 DNA Artificial Sequence <220> <223> Primer D8-6A <400> 59 gacctgttgg aaggacagac acaacgccca tcattctgct accaacgttc agggtcacga 60 tcccgacatt gataacctgc ctcccctcgc ctggtccgag g <210> <211> <212> <213> 60 101 DNA Artificial Sequence <220 <223> Primer D8-6B <400> 60 tcgtcctcgg accaggcgag gggaggcagg ttatcaatgt cgggatcgtg accctgaacg 60 ttggtagcag aatgatgggc gttgtgtctg tccttccaac a <210> <211> <212> <213> 61 95 DNA Artificial Sequence <220> <223> Primer D8-7A <400> 61 tcactcgagc ttctcccatc tcccgaaagc tcattcagtt ccaacagtac tatttcctgg 60 tcatctgtat tctcctgcga ttcatctggt gtttc <210> <211> <212> <213> 62 95 DNA Artificial Sequence <220> <223> Primer D8-7B <400> 62 actggaaaca ccagatgaat cgcaggagaa tacagatgac caggaaatag tactgttgga 60 actgaatgag ctttcgggag atgggagaag ctcga <210> <211> <212> <213> 63 89 DNA Artificial Sequence <220> <223> Primer D8-8A <400> 63 cagtgcgtgc tgaccgttcg atccctcaag gaccgagaca accagttcta cccgatctcag tacaagaaag aggccattgg actcgctct <210> <211> <212> <213> 64 89 DNA Artificial Sequence <220> <223> Primer D8-8B <400> 64 gtgcagagcg agtccaatgg cctctttctt gtactgagat cggtagaact ggttgtctcg gtccttgagg gatcgaacgg tcagcacgc <210> <211> <212> <213> 65 85 DNA Artificial Sequence <220> <223> Primer D8-9A <400> 65 gcactggact ctcaaggctc tgttccacct cttctttatg ccctccatcc tgacctcgct 60 cctggtgttc tttgtttccg agctc <210> <211> <212> <213> 66 85 DNA Artificial Sequence <220> <223> Primer D8-9B <400> 66 cgacgagctc ggaaacaaag aacaccagga gcgaggtcag gatggagggc ataaagaaga 60 ggtggaacag agccttgaga gtcca <210> 67 <211> 91 <212> DNA <213> Artificial Sequence <220> <223> <400> Primer D8-10A gtcggtggct tcggaattgc catcgtggtc ttcatgaacc actaccctct ggagaagatc 60 ggtgatcccg tctgggacgg acatggcttc t <210> <211> <212> <213> 68 91 DNA Artificial Sequence <220> <223> Primer D8-10B <400> 68 acagagaagc catgtccgtc ccagacggga tcaccgatct tctccagagg gtagtggttc 60 atgaagacca cgatggcaat tccgaagcca c <210> <211> <212> <213> 69 92 DNA Artificial Sequence <220> <223> Primer D8-11A <400> 69 ctgtgggtca gatecatgag accatgaaca ttegaegagg catcattact gactggttct 60 ttggaggcct gaactaccag atcgagcacc at <210> <211> <212> <213> 70 92 DNA Artificial Sequence <220> <223> Primer D8-11B <400> 70 agagatggtg ctcgatctgg tagttcaggc ctccaaagaa ccagtcagta atgatgcctc 60 gtcgaatgtt catggtctca tggatctgac cc <210> 71 <211> 93 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-12A <400> 71 ctctggccca ctctgcctcg acacaacctc actgccgttt cctaccaggt ggaacagctg 60 tgccagaagc acaacctccc ctaccgaaac cct <210> <211> <212> <213> 72 93 DNA Artificial Sequence <220> <223> Primer D8-12B <400> 72 gcagagggtt tcggtagggg aggttgtgct tctggcacag ctgttccacc tggtaggaaa 60 cggcagtgag gttgtgtcga ggcagagtgg gcc <210> <211> <212> <213> 73 90 DNA Artificial Sequence <220> <223> Primer D8-13A <400> 73 ctgccccatg aaggtctcgt catcctgctc cgatacctgg ccgtgttcgc tcgaatggcc gagaagcagc ccgctggcaa ggctctctaa <210> <211> <212> <213> 74 90 DNA Artificial Sequence <220> <223> Primer D8-13B <400> 74 ccgcttagag agccttgcca gcgggctgct tctcggccat tcgagcgaac acggccaggt 60 atcggagcag gatgacgaga ccttcatggg <210> <211> <212> <213> 75 38 DNA Artificial Sequence <223> Primer D8-1F <400> 75 tttccatgga gtccaagcga caggctctgt ctccccctc 38 <210> 76 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-3R <400> 76 tttagatctt gtacgagtac cagaggggag aggcgtc <210> 77 <211> 41 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-4F <400> 77 acaagatctc caccaccctg ggtcttggcg tgcttggata c 41 <210> 78 <211> 43 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-6R <400> 78 tttctcgagt gacatcgtcc tcggaccagg cgaggggagg cag 43 <210> 79 <211> 32 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-7F <400> 79 tcactcgagc ttctcccatc tcccgaaagc tc 32 <210> 80 <211> 29 <212> DNA <213> Artificial Sequence <223> Primer D8-9R <400> 80 cgacgagctc ggaaacaaag aacaccagg 29 <210> 81 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-10F <400> 81 tttgagctcg tcggtggctt cggaattgcc atcgtggtc 39 <210> 1 82 <210> > 36 <212> DNA <213> Artificial Sequence <220> <223> Primer D8-13R <400> 82 tttgcggccg cttagagagc cttgccagcg ggctgc 36 <210> 83 <211> 309 <212> DNA <213> Artificial Sequence <220> <223> fragmento Ncol / Bglll 309 bp de pT8(l- <400> 83 catggagtcc aagcgacagg ctctgtctcc cctccagctg 60 cgtgaacttc caccctggtg gagctgaaat cattgagaac 120 tgacgccttc atggttatgc actttcagga agccttcgac , 180 gatcaacccc tcctttgagc tgcctcccca ggctgccgtc ; 240 ccgaaagctc cgagaagagc tgatcgccac tggcatgttt < 300 ctcgtacaa 309 <210 > 84 <211> 321 <212> DNA <213> Artificial Sequence atggaacaga taccagggac aagctcaagc aacgaagctc gacgcctctc 3) cctacgacgt gagatgctac gaatgcccaa aggaggattt ccctctggta <223> Bge II / Xhol fragment 321 bp from pT8(4-6) <400> 84 gatctccacc accctgggtc ttggcgtgct tggatacttc ctgatggtcc agtaccagat 60 gtacttcatt ggtgctgtgc tgctcggtat gcactaccag caaatgggat ggctgtctca 120 tgacatctgc caccaccaga ccttcaagaa ccgaaactgg aataacctcg tgggtctggt 180 ctttggcaac ggactccagg gcttctccgt gacctgttgg aaggacagac acaacgccca 240 tcattctgct accaacgttc agggtcacga tcccgacatt gataacctgc ctcccctcgc 300 ctggtccgag gacgatgtca c 321 <210> 85 <211> 264 <212> DNA <213> Artificial Sequence <220> <223> fragmento Xhol / SacI 264 bp de pT8 (7-9) <400> 85 tcgagcttct cccatctccc gaaagctcat tcagttccaa cagtactatt tcctggtcat 60 ctgtattctc ctgcgattca tctggtgttt ccagtgcgtg ctgaccgttc gatccctcaa 120 ggaccgagac aaccagttct accgatctca gtacaagaaa gaggccattg gactcgctct 180 gcactggact ctcaaggctc tgttccacct cttctttatg ccctccatcc tgacctcgct 240 cctggtgttc tttgtttccg agct 264 <210> 86 <211> 369 <212> DNA <213> Artificial Sequence <220> <223> fragmento Sacl / Notl 309bp de pT8(l-3) <400> 86 cgtcggtggc ttcggaattg ccatcgtggt cttcatgaac cactaccctc tggagaagat 60 cggtgatccc gtctgggacg gacatggctt ctctgtgggt cagatccatg agaccatgaa 120 cattcgacga ggcatcatta ctgactggtt ctttggaggc ctgaactacc agatcgagca 180 ccatctctgg cccaccctgc ctcgacacaa cctcactgcc gtttcctacc aggtggaaca 240 gctgtgccag aagcacaacc tcccctaccg aaacctctg ccccatgaag gtctcgtcat 300 cctgctccga tacctggccg tgttcgctcg aatggccgag aagcagcccg ctggcaaggc 360 tctctaagc 369 <210> 87 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> Primer ODMW390 <400>87 aagaatcatt caccatgaag tccaagccgac aggc <210> 88 <211> 34 <212> DNA <213> Artificial Sequence <220> <223> Primer ODMW391 <400>88 gcctgtcgct tggacttcat ggtgaatgat tctt 34 <210> 89 <211> 1852 <212> DNA <213> Artificial Sequence <220> <223> chimeric terminator gene D85-1:XPR <400>89 cgatcaggag agaccgggtt ggcggcgtat ttgtgtccca aaaaacagcc ccaattgccc caattgaccc caaattgacc cagtagcggg cccaaccccg gcgagagccc ccttcacccc 120 acatatcaaa cctcccccgg ttcccacact tgccgttaag ggcgtagggt actgcagtct ggaatctacg cttgttcaga ctttgtacta gtttctttgt ctggccatcc gggtaaccca tgccggacgc aaaatagact actgaaaatt tttttgcttt gtggttggga ctttagccaa gggtataaaa gaccaccgtc cccgaattac ctttcctctt cttttctctc tctccttgtc aactcacacc cgaaatcgtt aagcatttcc ttctgagtat aagaatcatt caccatggag tccaagcgac aggctctgtc tcccctccag ctgatggaac agacctacga cgtcgtgaac ttccaccctg gtggagctga aatcattgag aactaccagg gacgagatgc tactgacgcc ttcatggtta tgcactttca ggaagccttc gacaagctca agcgaatgcc caagatcaac cccctcctttg agctgcctcc ccaggctgcc gtcaacgaag ctcaggagga tttccgaaag ctccgagaag agctgatcgc cactggcatg tttgacgcct ctccccctctg gtactcgtac aagatctcca ccaccctggg tcttggcgtg cttggatact tcctgatggt ccagtaccag atgtacttca 840 ttggtgctgt gctgctcggt atgcactacc agcaaatggg atggctgtct catgacatct 900 gccaccacca gaccttcaag aaccgaaact ggaataacct cgtgggtctg gtctttggca 960 acggactcca gggcttctcc gtgacctgtt ggaaggacag acacaacgcc catcattctg 1020 ctaccaacgt tcagggtcac gatcccgaca ttgataacct gcctcccctc gcctggtccg 1080 aggacgatgt cactcgagct tctcccatct cccgaaagct cattcagttc caacagtact 1140 atttcctggt catctgtatt ctcctgcgat tcatctggtg tttccagtgc gtgctgaccg 1200 ttcgatccct caaggaccga gacaaccagt tctaccgatc tcagtacaag aaagaggcca 1260 ttggactcgc tctgcactgg actctcaagg ctctgttcca cctcttcttt atgccctcca 1320 tcctgacctc gctcctggtg ttctttgttt ccgagctcgt cggtggcttc ggaattgcca tcgtggtctt catgaaccac taccctctgg agaagatcgg tgatcccgtc 1380 tgggacggac 1440 atggcttctc tgtgggtcag atccatgaga ccatgaacat tcgacgaggc atcattactg 1500 actggttctt tggaggcctg aactaccaga tcgagcacca tctctggccc accctgcctc 1560 gacacaacct cactgccgtt tcctaccagg tggaacagct gtgccagaag cacaacctcc cctaccgaaa ccctctgccc catgaaggtc tcgtcatcct gctccgatac 1620 ctggccgtg t 1680 tcgctcgaat ggccgagaag cagcccgctg gcaaggctct ctaagcggcc gccaccgccg 1740 agattccggc ctcttcggcc gccaagcgac ccgggtggac gtctagaggt acctagcaat 1800 taacagatag tttgccggtg ataattctct taacctccca cactcctttg acataacgat ttatgtaacg aaactgaaat ttgaccagat attgtgtccg eg <210> 90 <211> 1898 <212> DNA <213> Artificial Sequence <220> <223> gene terminado: r quimérico D8S-2:XPR <400> 90 cgatcaggag agaccgggtt ggeggegtat ttgtgtccca aaaaacagcc ccaattgccc 60 caattgaccc caaattgacc cagtagcggg cccaaccccg gcgagagccc ccttcacccc 120 acatatcaaa cctcccccgg ttcccacact tgccgttaag ggcgtagggt aetgeagtet 180 ggaatctacg cttgttcaga ctttgtacta gtttctttgt ctggccatcc gggtaaccca 240 tgccggacgc aaaatagact aetgaaaatt tttttgcttt gtggttggga ctttagccaa 300 gggtataaaa gaccaccgtc cccgaattac ctttcctctt cttttctctc tctccttgtc aactcacacc cgaaatcgtt aagcatttcc 420 tccaagcgac 480 aggctctgtc tcccctccag ttccaccctg 540 gtggagctga aatcattgag ttcatggtta 600 tgcactttca ggaagccttc ccctcctttg 660 agctgcctcc ccaggctgcc ctccgagaag 720 agctgatcgc cactggcatg aagatctcca 780 ccaccctggg tcttggcgtg atgtacttca 840 ttggtgctgt gctgctcggt catgacatct 900 gccaccacca gaccttcaag gtctttggca 960 acggactcca gggcttctcc catcattctg 1020 ctaccaacgt tcagggtcac gcctggtccg 1080 aggacgatgt cactcgagct caacagtact 1140 atttcctggt catctgtatt gtgctgaccg 1200 ttcgatccct caaggaccga aaagaggcca 1260 ttggactcgc tctgcactgg atgccctcca 1320 tcctgacctc gctcctggtg ggaattgcca 1380 tcgtggtctt catgaaccac tgggacggac 1440 atggcttctc tgtgggtcag atcattactg 1500 actggttctt tggaggcctg accctgcctc 1560 gacacaacct cactgccgtt cacaacctcc 1620 cctaccgaaa ccctctgccc ctggccgtgt 1680 tcgctcgaat ggccgagaag gccaccgcgg 1740 cccgagattc cggcctcttc aggtacctag 1800 caattaacag atagtttgcc tttgacataa 1860 cgatttatgt aacgaaactg agctccagct 1898 tttgttccct ttagtgaggg ttctgagtat aagaatcatt caccatgaag ctgatggaac agacctacga cgtcgtgaac aactaccagg gacgagatgc tactgacgcc gacaagctca agcgaatgcc caagatcaac gtcaacgaag ctcaggagga tttccgaaag tttgacgcct ctcccctctg gtactcgtac cttggatact tcctgatggt ccagtaccag atgcactacc agcaaatggg atggctgtct aaccgaaact ggaataacct cgtgggtctg gtgacctgtt ggaaggacag acacaacgcc gatcccgaca ttgataacct gcctcccctc tctcccatct cccgaaagct cattcagttc ctcctgcgat tcatctggtg tttccagtgc gacaaccagt tctaccgatc tcagtacaag actctcaagg ctctgttcca cctcttcttt ttctttgttt ccgagctcgt cggtggcttc taccctctgg agaagatcgg tgatcccgtc atccatgaga ccatgaacat tcgacgaggc aactaccaga tcgagcacca tctctggccc tcctaccagg tggaacagct gtgccagaag catgaaggtc tcgtcatcct gctccgatac cagcccgctg gcaaggctct ctaagcggcc ggccgccaag cgacccgggt ggacgtctag ggtgataatt cctcttaacct cccacactcc aaatttgacc agatattgtg tccgcggtgg ttaattaa <210> 91 <211> 45 <212> DNA <213> Artificial Sequence <223> Primer ODMW392 <400> 91 gaacagacct aggacgtctc cgcttgggtg aacttccacc ctggt <210> 92 <211> 45 <212> DNA <213> Artificial Sequence <220> <223> Primer ODMW393 <400> 92 accagggtgg aagttcaccc aagcggagac gtcgtaggtc tgttc <210> 93 <211> 1269 <212> DMA <213> Artificial Sequence <220> <223> deltas 8 gene - codon-optimized synthetic desaturase for Yarrowia lipolytica in pDMW2651 <400> 93 atgaagtcca agcgacaggc tctgtctccc ctccagctga tggaacagac ctacgacgtc 60 tccgcttggg 120 tgaacttcca ccctggtgga gctgaaatca ttgagaacta ccagggacga gatgctactg 180 acgccttcat ggttatgcac tttcaggaag ccttcgacaa gctcaagcga atgcccaaga 240 tcaacccctc ctttgagctg cctccccagg ctgccgtcaa cgaagctcag gaggatttcc 300 gaaagctccg agaagagctg atcgccactg gcatgtttga cgcctctccc ctctggtact 360 cgtacaagat ctccaccacc ctgggtcttg gcgtgcttgg atacttcctg atggtccagt 420 accagatgta cttcattggt gctgtgctgc tcggtatgca ctaccagcaa atgggatggc 480 tgtctcatga catctgccac caccagacct tcaagaaccg aaactggaat aacctcgtgg 540 gtctggtctt tggcaacgga ctccagggct tctccgtgac ctgttggaag gacagacaca 600 acgcccatca ttctgctacc aacgttcagg gtcacgatcc cgacattgat aacctgcctc 660 ccctcgcctg gtccgaggac gatgtcactc gagcttctcc catctcccga aagctcattc 720 agttccaaca gtactatttc ctggtcatct gtattctcct gcgattcatc tggtgtttcc 780 agtgcgtgct gaccgttcga tccctcaagg accgagacaa ccagttctac cgatctcagt 840 acaagaaaga ggccattgga ctcgctctgc actggactct caaggctctg ttccacctct 900 tcttta tgcc ctccatcctg acctcgctcc tggtgttctt tgtttccgag ctcgtcggtg gcttcggaat tgccatcgtg gtcttcatga accactaccc tctggagaag 960 atcggtgatc ccgtctggga cggacatggc ttctctgtgg gtcagatcca tgagaccatg 1020 aacattcgac gaggcatcat tactgactgg ttctttggag gcctgaacta ccagatcgag 1080 caccatctct ggcccaccct gcctcgacac aacctcactg ccgtttccta ccaggtggaa 1140 cagctgtgcc agaagcacaa cctcccctac cgaaaccctc tgccccatga aggtctcgtc 1200 atcctgctcc gatacctggc cgtgttcgct cgaatggccg agaagcagcc cgctggcaag 1260 gctctctaa <210> 94 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> Primer ODMW404 <400> 94 cctggtacca tgaagtccaa gccacaggc <210> 95 <211> 1272 <212> DNA <213> Artificial Sequence <220> <223> chimeric gene <400> 95 catgaagtcc aagcgacagg ctctgtctcc cctccagctg atggaacaga cctacgacgt 60 ctccgcttgg gtgaacttcc accctggtgg agctgaaatc attgagaact accagggacg 120 agatgctact gacgccttca tggttatgca ctttcaggaa gccttcgaca agctcaagcg 180 aatgcccaag atcaacccct cctttgagct gcctccccag gctgccgtca acgaagctca 240 ggaggatttc cgaaagctcc gagaagagct gatcgccact ggcatgtttg acgcctctcc 300 cctctggtac tcgtacaaga tctccaccac cctgggtctt ggcgtgcttg gatacttcct 360 gatggtccag taccagatgt acttcattgg tgctgtgctg ctcggtatgc actaccagca 420 aatgggatgg ctgtctcatg acatctgcca ccaccagacc ttcaagaacc gaaactggaa 480 taacctcgtg ggtctggtct ttggcaacgg actccagggc ttctccgtga cctgttggaa 540 ggacagacac aacgcccatc attctgctac caacgttcag ggtcacgatc cccgacattga 600 taacctgcct cccctcgcct ggtccgagga cgatgtcact cgagcttctc ccatctcccg aaagctcatt 720 ctggtgtttc 780 cccgatctcag 840 gttccacctc 900 gctcgtcggt 960 gatcggtgat 1020 gaacattcga 1080 gcaccatctc 1140 acagctgtgc 1200 catcctgctc 1260 ggctctctctaa 1272 cagttccaac cagtgcgtgc tacaagaaag ttctttatgc ggcttcggaa cccgtctggg cgaggcatca tggcccaccc cagaagcaca cgatacctgg gc agtactattt tgaccgttcg aggccattgg cctccatcct ttgccatcgt aggcatgg ttactgactg tgcctcgaca accctccccta ccgtgttcgc cctggtcatc atccctcaag actcgctctg gacctcgctc ggtcttcatg cttctctgtg gttctttgga caacctcact ccgaaaccct tcgaatggcc tgtattctcc gaccgagaca cactggactc ctggtgttct aaccactacc ggtcagatcc ggcctgaact gccgtttcct ctgccccatg gagaagcagc tgcgattcat accagttcta tcaaggctct ttgtttccga ctctggagaa atgagaccat accagatcga accaggtgga aaggtctcgt ccgctggcaa <210> 96 <211> 80 <212> DNA <213> Artificial Sequence <220> <223> Primer YL521 <400> 96 tttccatggt gaagtccaag cgacaggctc tgcccctcac catcgacgga actacctacg 60 acgtctccgc ttgggtgaac <210> <211> <212> <213> 97 30 DNA Artificial Sequence <220> <223> Primer YL522 <400> 97 tggagatctt gtacgagtac cagaggggag <210> 98 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> Primer YL525 <400> 98 ccttcatggt tatgcactct caggaagcct tcgacaa 37 <210> <211> <212> <213> 99 37 DNA Artificial Sequence <220> <223> Primer YL526 <400> 99 ttgtcgaagg cttcctgaga gtgcataacc atgaagg 37 <210> <211> <212> <213> 100 38 DNA Artificial Sequence <220> <223> Primer YL527 <400> 100 ccaagatcaa ccctcctcc gagctgcctc cccaggct 38 <210> <211> <212> <213> 101 38 DNA Artificial Sequence <220> <223> Primer YL528 <400> 101 agcctgggga ggcagctcgg aggaggggtt gatcttgg 38 <210> <211> <212> <213> 102 37 DNA Artificial Sequence <220> <223> Primer YL529 <400> 102 gggcttctcc gtgacctggt ggaaggacag acacaac 37 <210> 103 <211> 37 <212> DNA <213> Artificial Sequence <220> <223> Primer YL530 <400> 103 gttgtgtctg tccttccacc aggtcacgga gaagccc <210> 104 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Primer YL531 <400> 104 acattgataa cctgcctctg ctcgcctggt cccgaggac <210> 105 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Primer YL532 <400> 105 gtcctcggac caggcgagca gaggcaggtt atcaatgt <210> 106 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Primer YL533 <400> 106 tcatctggtg tttccagtct gtgctgaccg ttcgatcc <210> 107 <211> 38 <212> DNA <213> Artificial Sequence <220> <223> Primer YL534 <400> 107 ggatcgaacg gtcagcacag actggaaaca ccagatga <210> 108 <211> 39 <212> DNA <213> Artificial Sequence <220> <223> Primer YL535 <400> 108 ctgcactgga ctctcaagac cctgttccac ctcttcttt 39 <210> <211> <212> <213> DNA Artificial Sequence <220> <223> Primer YL536 <400> 109 aaagaagagg tggaacaggg tcttgagagt ccagtgcag 39 <210> <211> <212> <213> 110 37 DNA Artificial Sequence <220> <223> Primer YL537 <400> 110 ctggagaaga tcggtgattc cgtctgggac ggacatg <210> <211> <212> <213> 111 37 DNA Artificial Sequence <220> <223> Primer YL538 <400> 111 catgtccgtc ccagacggaa tcaccgatct tctccag 37 <210> <211> <212> <213> 112 1272 DNA Artificial Sequence 220> <223> Synthetic delta-8 desaturase (codon-optimized for Yarrowia lipolytics) <400> 112 catggtgaag 60 tccaagcgac aggctctgcc cctcaccatc gacggaacta cctacgacgt ctccgcttgg 120 gtgaacttcc accctggtgg agctgaaatc attgagaact accagggacg agatgctact 180 gacgccttca tggttatgca ctctcaggaa gccttcgaca agctcaagcg aatgcccaag 240 atcaacccct cctccgagct gcctccccag gctgccgtca acgaagctca ggaggatttc 300 cgaaagctcc gagaagagct gatcgccact ggcatgtttg acgcctctcc cctctggtac 360 tcgtacaaga tctccaccac cctgggtctt ggcgtgcttg gatacttcct gatggtccag 420 taccagatgt acttcattgg tgctgtgctg ctcggtatgc actaccagca aatgggatgg 480 ctgtctcatg acatctgcca ccaccagacc ttcaagaacc gaaactggaa taacctcgtg 540 ggtctggtct ttggcaacgg actccagggc ttctccgtga cctggtggaa ggacagacac 600 aacgcccatc attctgctac caacgttcag ggtcacgatc ccgacattga taacctgcct 660 ctgctcgcct ggtccgagga cgatgtcact cgagcttctc ccatctcccg aaagctcatt 720 cagttccaac agtactattt cctggtcatc tgtattctcc tgcgattcat ctggtgtttc 780 cagtctgtgc tgaccgttcg atccctcaag gaccgagaca accagttcta ccgatctcag 840 tacaagaaag aggccattgg actcgctctg cactggactc tcaagaccct gttccacctc 900 ttcttt atgc cctccatcct gacctcgctc ctggtgttct ttgtttccga gctcgtcggt 960 ggcttcggaa ttgccatcgt ggtcttcatg aaccactacc ctctggagaa gatcggtgat 1020 tccgtctggg acggacatgg cttctctgtg ggtcagatcc atgagaccat gaacattcga 1080 cgaggcatca ttactgactg gttctttgga ggcctgaact accagatcga gcaccatctc 1140 tggcccaccc tgcctcgaca caacctcact gccgtttcct accaggtgga acagctgtgc 1200 cagaagcaca acctccccta ccgaaaccct ctgccccatg aaggtctcgt catcctgctc 1260 cgatacctgg ccgtgttcgc tcgaatggcc gagaagcagc ccgctggcaa ggctctctaa gc 1272 <210> 113 <211> 422 <212> PRT <213> Artificial Sequence <220> <223> Synthetic delta-8 desaturase (codon-optimized for Yarrowia lipolytica) <400> 113 Met Vai Lys Ser Lys Arg Gin Ala Leu Pro Leu Thr lie Asp Gly Thr 15 10 15 Thr Tyr Asp Vai 20 Ser Ala Trp Vai Asn Phe 25 His Pro Gly Gly 30 Ala Glu lie He Glu Asn Tyr Gin Gly Arg Asp Ala Thr Asp Ala Phe Met Vai 35 40 45 Met His Ser Gin Glu Ala Phe Asp Lys Leu Lys Arg Met Pro Lys He 50 55 60 Asn Pro Ser Ser Glu Leu Pro Gin Ala Vai Asn Glu Ala Gin 65 70 75 80 Glu Asp Phe Arg Lys Leu Arg Glu Glu Leu He Ala Thr Gly Met Phe 85 90 95 Asp Ala Ser Pro Leu Trp Tyr Ser Tyr Lys lie Ser Thr Thr Leu Gly 100 105 110 Leu Gly Vai Leu Gly Tyr Phe Leu Met Vai Gin Tyr Gin Met Tyr Phe 115 120 125 He Gly Ala Vai Leu Leu Gly Met His Tyr Gin Gin Met Gly Trp Leu 130 135 14 0 Ser His Asp He Cys His His Gin Thr Phe Lys Asn Arg Asn Trp Asn 145 150 155 160 Asn Leu Vai Gly Leu Vai Phe Gly Asn Gly Leu Gin Gly Phe Ser Vai 165 170 175 Thr Trp Trp Lys Asp Arg His Asn Ala His His Ser Ala Thr Asn Vai 180 185 190 Gin Gly His Asp Pro Asp lie Asp Asn Leu Pro Leu Leu Ala Trp Ser 195 200 205 Glu Asp Asp Vai Thr Arg Ala Ser Pro lie Ser Arg Lys Leu He Gin 210 215 220 Phe Gin Gin Tyr Tyr Phe L Eu Vai He Cys He Leu Leu Arg Phe He 225 230 235 240 Trp Cys Phe Gin Ser Vai Leu Thr Vai Arg Ser Leu Lys Asp Arg Asp 245 250 255 Asn Gin Phe Tyr Arg Ser Gin Tyr Lys Lys Glu Ala He Gly Leu Ala 260 265 270 Leu His Trp Thr Leu Lys Thr Leu Phe His Leu Phe Phe Met Pro Ser 275 280 285 He Leu Thr Ser Leu Vai Phe Phe Vai Ser Glu Leu Vai Gly Gly 290 295 300 Phe Gly lie Ala He Vai Vai Phe Met Asn His Tyr Pro Leu Glu Lys 305 310 315 320 lie Gly Asp Ser Vai Trp Asp Gly His Gly Phe Ser Vai Gly Gin lie 325 330 335 His Glu Thr Met Asn 340 He Arg Arg Gly 345 He He Thr Asp Trp 350 Phe Phe Gly Gly Leu Asn Tyr Gin He Glu His His Leu Trp Pro Thr Leu Pro 355 360 365 Arg His Asn Leu Thr Ala Vai Ser Tyr Gin Vai Glu Gin Leu Cys Gin 370 375 380 Lys His Asn Leu Pro Tyr Arg Asn Pro Leu Pro His Glu Gly Leu Vai 385 390 395 400 lie Leu Leu Arg Tyr Leu Ala Vai Phe Ala Arg Met Ala Glu Lys Gin 405 410 415 Pro Wing Gly Lys Wing Leu <210> 114 <211> 995 <212> DNA <213> Yarrowia lipolytica <400> 114 agtgtacgca gtactataga ggaacaattg ccccggagaa gacggccagg ccgcctagat 60 gacaaattca acaactcaca gctgactttc tgccattgcc actagggggg ggccttttta 120 tatggccaag ccaagctctc cacgtcggtt gggctgcacc caacaataaa tgggtagggt 180 tgcaccaaca aagggatggg atggggggta gaagatacga ggataacggg gctcaatggc 240 acaaataaga acgaatactg ccattaagac tcgtgatcca gcgactgaca ccattgcatc 300 atctaagggc ctcaaaacta cctcggaact gctgcgctga tctggacacc acagaggttc 360 cgagcacttt aggttgcacc aaatgtccca ccaggtgcag gcagaaaacg ctggaacagc 420 gtgtacagtt tgtcttaaca aaaagtgagg gcgctgaggt cgagcagggt ggtgtgactt 480 gttatagcct ttagagctgc gaaagcgcgt atggatttgg ctcatcaggc cagattgagg 540 gtctgtggac acatgtcatg ttagtgtact tcaatcgccc cctggatata gccccgacaa 600 taggccgtgg cctcattttt ttgccttccg cacatttcca ttgctcggta cccacacctt 660 gcttctcctg cacttgccaa ccttaatact ggtttacatt gaccaacatc ttacaagcgg 720 ggggcttgtc tagggtatat ataaacagtg gctctcccaa tcggttgcca gtctcttttt 780 tcctttcttt ccccacagat tcgaaatcta aactacacat cacacaatgc ctgttactga 840 cgtccttaag cgaaagtccg gtgtcatcgt cggcgacgat gtccgagccg tgagtatcca 900 cgacaagatc agtgtcgaga cgacgcgttt tgtgtaatga cacaatccga aagtcgctag 960 caacacacac tctctacaca aactaaccca gctct 995 <210> 115 <211> 8502 <212> DNA <213> Artificial Sequence <220> <223> Plasmid pY54PC <400> 115 ggccgccacc 60 gcggcccgag attccggcct cttcggccgc caagcgaccc gggtggacgt ctagaggtac 120 ctagcaatta acagatagtt tgccggtgat aattctctta acctcccaca ctcctttgac 180 ataacgattt atgtaacgaa actgaaattt gaccagatat tgtgtccgcg gtggagctcc 240 agcttttgtt ccctttagtg agggttaatt aatcgagctt ggcgtaatca tggtcatagc 300 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 360 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 420 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 480 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 540 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 600 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 660 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 720 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 780 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 840 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 900 gtaggt atct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 960 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 1020 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 1080 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 1140 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 1200 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 1260 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 1320 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 1380 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 1440 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 1500 cgggagggct 1560 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 1620 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 1680 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 1740 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 1800 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 1860 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 1920 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 1980 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 2040 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 2100 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 2160 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 2220 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 2280 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattat tgaa 2340 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 2400 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg 2460 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg 2520 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc 2580 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc 2640 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga 2700 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa 2760 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga 2820 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 2880 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 2940 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3000 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3060 attgtaatac gactcactat agggcgaatt gggtaccggg ccccccctcg aggtcgacgg 3120 tatcgataag cttgatatcg aattcatgtc acacaaaccg atcttcgcct caaggaaacc 3 180 taattctaca tccgagagac tgccgagatc cagtctacac tgattaattt tcgggccaat 3240 catgatgata 3300 aactgatgtt 3360 tctaccgcct 3420 tattattaga 3480 atggcagttc 3540 tcttaaatat 3600 aaaaaaatcc 3660 tattcacacg 3720 tctcttctag 3780 ttcatcccac 3840 acaattataa 3900 gtgcttctcg 3960 tatataatcc 4020 ttgcttaaat 4080 ttttgaagaa 4140 gcagaatcta 4200 gagatattgt 4260 ttgatgcatc 4320 ttaccgctat 4380 catagactta 4440 ttgggtgtaa 4500 ataatttgaa 4560 gagaggtctg 4620 cactgaatgt 4680 cgtctaacgg 4740 atcatttgta 4800 gtccacccct 4860 caccactaaa 4920 aatttaaaaa ctgacagtca ctcaatattt ccaaatgatg caacttactt gttcatttaa ggatagcata cttgtacaac ttactattga aaatacaggt atattccttg taagatatac tatttatttt ttttgtttat tcaatccccc gcaaaaaaaa gaatgcggta acatttttgc cacaacagtt gtatacctac tgaatctgca tattgggatc tcgaatcgga tgactgtcag agtgtacctc acttgatata acaattaccc ttccaaattg accccacaaa tataatatta ctaaatagac ctcgcattgt atattgtatg aaacacttcc agaataaatg gcattgccta atcgagaaat ggacgagaat actattctca caatgaatga gtatagtggc atccattaaa ggatacataa gtcaactgta aaaaaatcgt tgttcttcga acaagtacat ttttttgttt agtaagccgg tgcgagttac tcaacggatg aacccgagta cttcattatg atacagtgcc aaacaaatga aaggtattga aactcatata ccgaatatac aatcgtgtta tgtcccattg aaggggtcat ttctcaaaat gctttatgaa caatttatgt aatgatatct ataaatagtc gattattatt acaagtatgt gatttctctc caaagtagcg tattctaatg tacatgggct ctcgttcagt tgaaagaaaa tgcggtacat ttttacaagt tgttttgttt ttgtacttgt cggtgtgcgc tgttcggaaa gcctaaaatg tacaaggtgc taaaaatgaa caaccaatta tgtacaaact tcatgcctac atatatctta tatgtattat agactccatc ttaataataa aacttatttt tatttaggaa ttataaatgc attcgaaatc atcaactatc cacacactca ttgttcatac cattctatct aatcaaaaag ggtatatatt aggtattttg atggtaggaa atttccaggt acgtaaaagt cgtacaacta tttttttttc gttattggcg ttttagctta ctcaaccgat tatctcataa ccctcaacct aaaagccaag aaagaaatac aatcccacaa ccaagcacta agtaacaagc atatatacat tgccgcctcc acagactcca tattacttag acaatttata gtatgggaaa aacagcaacg aaagaacagc actgtctttc ttctagtcat tgcaaattca cttctctggt tatttcttgt atttaatttt attaccatac tagacgttcc tgcgctccct tgtactactg taatgattca ttcaattaat tgcatgctac ttcgacagta aattctcggt taccatacct gcactgagct agttctttgt tattcccaaa acctaccaaa taccaccaca ctcgttgggt gcagtcgcca gcttaaagat atctatccac atcagccaca actcccttcc 4980 tttaataaac 5040 cgactacacc cttggctatt gaggttatga gtgaatatac tgtagacaag acactttcaa 5100 gaagactgtt tccaaaacgt accactgtcc tccactacaa acacacccaa tctgcttctt 5160 ctagtcaagg ttgctacacc ggtaaattat aaatcatcat ttcattagca gggcagggcc 5220 ctttttatag agtcttatac actagcggac cctgccggta gaccaacccg caggcgcgtc 5280 agtttgctcc ttccatcaat gcgtcgtaga aacgacttac tccttcttga gcagctcctt 5340 gaccttgttg gcaacaagtc tccgacctcg gaggtggagg aagagcctcc gatatcggcg 5400 gtagtgatac cagcctcgac ggactccttg acggcagcct caacagcgtc accggcgggc 5460 ttcatgttaa gagagaactt gagcatcatg gcggcagaca gaatggtggc aatggggttg 5520 accttctgct tgccgagatc gggggcagat ccgtgacagg gctcgtacag accgaacgcc 5580 tcgttggtgt cgggcagaga agccagagag gcggagggca gcagacccag agaaccgggg 5640 atgacggagg cctcgtcgga gatgatatcg ccaaacatgt tggtggtgat gatgatacca 5700 ttcatcttgg agggctgctt gatgaggatc atggcggccg agtcgatcag ctggtggttg 5760 agctcgagct gggggaattc gtccttgagg actcgagtga cagtctttcg ccaaag tcga 5820 gaggaggcca gcacgttggc cttgtcaaga gaccacacgg gaagaggggg gttgtgctga 5880 agggccagga aggcggccat tcgggcaatt cgctcaacct caggaacgga gtaggtctcg 5940 gtgtcggaag cgacgccaga tccgtcatcc tcctttcgct ctccaaagta gatacctccg 6000 acgagctctc ggacaatgat gaagtcggtg ccctcaacgt ttcggatggg ggagagatcg 6060 gcgagcttgg gcgacagcag ctggcagggt cgcaggttgg cgtacaggtt caggtccttt 6120 cgcagcttga ggagaccctg ctcgggtcgc acgtcggttc gtccgtcggg agtggtccat 6180 acggtgttgg cagcgcctcc gacagcaccg agcataatag agtcagcctt tcggcagatg 6240 tcgagagtag cgtcggtgat gggctcgccc tccttctcaa tggcagctcc tccaatgagt 6300 cggtcctcaa acacaaactc ggtgccggag gcctcagcaa cagacttgag caccttgacg 6360 gcctcggcaa tcacctcggg gccacagaag tcgccgccga gaagaacaat cttcttggag 6420 tcagtcttgg tcttcttagt ttcgggttcc attgtggatg tgtgtggttg tatgtgtgat 6480 gtggtgtgtg gagtgaaaat ctgtggctgg caaacgctct tgtatatata cgcacttttg 6540 cccgtgctat gtggaagact aaacctccga agattgtgac tcaggtagtg cggtatcggc 6600 tagggaccca aaccttgtcg atgccgatag cgctatcgaa cgtaccccag ccggccggga 6 660 gtatgtcgga ggggacatac gagatcgtca agggtttgtg gccaactggt aaataaatga 6720 agagaccggg 6780 cccaaattga 6840 aacctccccc 6900 cgcttgttca 6960 gcaaaataga 7020 aagaccaccg 7080 cccgaaatcg 7140 cagtgtgagg 7200 gaaggatgcc 7260 gttcgtccct 7320 tgacgtcttt 7380 tggtgatatt 7440 caagctgcgt 7500 cttcaaggtc 7560 gggccagacc 7620 gcagtgcgga 7680 gggtgatctt 7740 gaaggacaag 7800 tgacacccac 7860 agatgaggag 7920 cttccccatt 7980 gcctaacggt 8040 gctgtcgctt 8100 tcccgtcaac 8160 gatcgtgttc 8220 tatggatttc 8280 caactggttc 8340 tcgccacaac 8400 tgactcaggc ttggcggcgt cccagtagcg ggttcccaca gactttgtac ctactgaaaa tccccgaatt ttaagcattt acgtttactc gaggcaccct gatcatcccg gacacttttc gacgagagcg accttgttcc tcgttcaacc tcgaccctcg tggttggctc ttcggcgcct cacaacactc cctctgttga ctgacccgca ctctcgtttg caggcccaca gcgatgcact atgctggtgt tcgctcaacc ttcacgaagc acgggtggat ttttcaaaga gacgacggaa atttgtgtcc ggcccaaccc cttgccgtta tagtttcttt tttttttgct acctttcctc ccttctgagt gggccgaggt tcttgatgat gtggaagtgt accccgaggc accgcgatat agtctcttgg tctgcatctg ccaacgtgct acgacttttt tcttgggagg accacgccgc cctggagtga tgtggtcgcg cccgtctctc agccctcggg ggacctggta actttttggt ac...

Claims

Claims 1. ISOLATED POLYNUCLEOTIDE, comprising: (a) a nucleotide sequence encoding a polypeptide that has delta-8 desaturase activity, wherein the polypeptide has an amino acid sequence consisting essentially of SEQ ID No. 2 or 113; or, (b) a complement of the nucleotide sequence, wherein the complement and the nucleotide sequence consist of the same number of nucleotides and are 100% complementary.

2. POLYNUCLEOTIDE, according to claim 1, characterized in that the nucleotide sequence comprises SEQ ID No.: 1 or 112.

3. POLYPEPTIDE, according to claim 1, characterized in that the amino acid sequence of the polypeptide comprises SEQ ID No.: 2 or 113.

4. RECOMBINANT CONSTRUCTION, comprising the polynucleotide as described in claim 1, operably linked to at least one regulatory sequence.

5. CELL, comprising the recombinant construct as described in claim 4.

6. CELL, according to claim 5, characterized in that the cell is selected from the group consisting of plants and yeast.

7. YARROWIA SP. TRANSFORMED, comprising the recombinant construct as described in claim 4.

8. YARROWIA SP. TRANSFORMED, according to claim 7, characterized in that it is selected from the group consisting of Yarrowia lipolytica ATCC 20362, Yarrowia lipolytica ATCC 8862, Yarrowia lipolytica ATCC 18944, Yarrowia lipolytica ATCC 76982 and Yarrowia lipolytica LGAM S(7)1.

9. METHOD FOR TRANSFORMING A CELL, comprising transforming a cell with the recombinant construct as described in claim 4 and selecting the cells transformed with the recombinant construct as described in claim 4.

10. METHOD FOR PRODUCING A TRANSFORMED PLANT, comprising transforming a plant cell with the polynucleotide as described in claim 1 and regenerating a plant from the transformed plant cell.

11. METHOD, according to claim 10, characterized in that the plant is a soybean plant.

12. METHOD FOR THE PRODUCTION OF YEAST, comprising transforming a yeast cell with the polynucleotide as described in claim 1 and developing yeast from the transformed yeast cell.

13. METHOD FOR THE PRODUCTION OF YEAST, comprising transforming a yeast cell with the polynucleotide as described in claim 1 and developing yeast from the transformed yeast cell, wherein the yeast is an oleaginous yeast selected from the group of: Yarrowia, Candida, Rhodotorula, Rhodosporidium, Cryptococcus, Trichosporon and Lipomyces.

14. SEED, comprising the recombinant construct as described in claim 4.

15. SEED, which is obtained from the plant produced by the method described in claim 10 or 11.

16. METHOD FOR THE PRODUCTION OF LONG-CHAIN ​​POLYUNSATURATED FATTY ACIDS IN A CELL, comprising: (a) the transformation of a cell with the recombinant construct as described in claim 4; (b) the selection of transformed cells that form long-chain polyunsaturated fatty acids.

17. OIL, which is obtained from the seed as described in claim 14.

18. OIL, which is obtained from the seed as described in claim 15.

19. OIL, which is obtained from yeast produced by the method described in claim 12 or 13.

20. METHOD FOR THE PRODUCTION OF LONG-CHAIN ​​POLYUNSATURATED FATTY ACIDS IN A PLANT CELL, comprising: (a) the transformation of a cell with the recombinant construct as described in claim 4; and (b) the selection of transformed cells that form long-chain polyunsaturated fatty acids.

21. METHOD FOR THE PRODUCTION OF AT LEAST ONE POLYUNSATURATED FATTY ACID in a soybean cell, comprising: (a) the transformation of a soybean cell with a first recombinant DNA construct comprising a single polynucleotide encoding a delta-8 desaturase polypeptide, operably linked to at least one regulatory sequence, and at least one additional recombinant DNA construct comprising a single polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of delta-4, delta-5, delta-6, delta-9, delta-12, delta-15 and delta-17 desaturase, delta-9 elongase, Cis elongase to C22 and C20 elongase to C 24 ; (b) the regeneration of a soybean plant from the transformed cell from step (a); and (c) the selection of seeds obtained from plants in step (b) that have an altered level of polyunsaturated fatty acids when compared to the level in seeds obtained from unprocessed soybean plants.

22. OILSEED PLANT, comprising the recombinant construct as described in claim 4.

23. OILSEED PLANT, comprising: (a) a first recombinant DNA construct comprising a single polynucleotide encoding a delta-8 desaturase polypeptide, operably linked to at least one regulatory sequence; and (b) at least one additional recombinant DNA construct comprising a single polynucleotide, operably linked to at least one regulatory sequence, encoding a polypeptide selected from the group consisting of delta-4, delta-5, delta-6, delta-9, delta-12, delta-15 and delta-17 desaturase, delta-9 elongase, elongase C 18 C22 and C2o elongase 24 .

24. PLANT, according to claim 22 or 23, characterized in that the oilseed plant is selected from the group consisting of soybean, Brassica species, sunflower, corn, cotton, flax and saffron.

25. PLANT, according to claim 22 or 23, characterized in that the oilseed plant is selected from the group consisting of soybean, Brassica species, sunflower, corn, cotton, flax and safflower, and in that the polyunsaturated fatty acid is selected from the group consisting of AA, EDA, EPA, ETA, EtrA, DGLA, DPA and DHA.

26. SEED, which is obtained from the plant as described in claim 22.

27. SEED, which is obtained from the plant as described in claim 23.

28. OIL, which is obtained from the seed as described in claim 26.

29. OIL, which is obtained from the seed as described in claim 27.

30. OIL, which is obtained by the method described in claim 20 or 21.

31. FOOD OR FEED, incorporating the oil as described in claim 28.

32. FOOD OR FEED, incorporating the oil as described in claim 29.

33. FOOD OR FEED, incorporating the oil as described in claim 30.

34. FOOD OR FEED, comprising the seeds as described in claim 31.

35. FOOD OR FEED, comprising the seeds as described in claim 32.

36. FOOD OR FEED, comprising the seeds as described in claim 33.

37. FOOD OR FEED, comprising an ingredient derived from the processing of seeds as described in claim 26.

38. FOOD OR FEED, comprising an ingredient derived from the processing of seeds as described in claim 27.