Abstract:
The present invention relates generally to novel genetic sequences that encode fatty acid epoxygenase enzymes, in particular fatty acid Δ12-epoxygenase enzymes from plants that are mixed function monooxygenase enzymes. More particularly, the present invention exemplifies cDNA sequences from Crepis spp. and  Vernonia galamensis  that encode fatty acid Δ12-epoxygenases. The genetic sequences of the present invention provide the means by which fatty acid metabolism may be altered or manipulated in organisms, such as, for example, yeasts, moulds, bacteria, insects, birds, mammals and plants, and more particularly in plants. The invention also extends to genetically modified oil-accumulating organisms transformed with the subject genetic sequences and to the oils derived therefrom. The oils thus produced provide the means for the cost-effective raw materials for use in the efficient production of coatings, resins, glues, plastics, surfactants and lubricants.

Description:
RELATED APPLICATION DATA  
       [0001]    This application is a continuation-in-part application of U.S. Ser. No. 09/059,769 filed on Apr. 14, 1998, which claims benefit of priority under Title 35, U.S.C. §119 from Australian Patent Application No. PO6223 filed on Apr. 15, 1997 and Australian Patent Application No. PO6226 filed on Apr. 15, 1997, and which also claims benefit of priority under Title 35, U.S.C. §119(e) from U.S. Ser. No. 60/043,706 filed on Apr. 16, 1997 and from U.S. Ser. No. 60/050,403 filed on Jun. 20,1997. 
     
    
     
       FIELD OF THE INVENTION  
         [0002]    The present invention relates generally to novel genetic sequences that encode fatty acid epoxygenase enzymes. In particular, the present invention relates to genetic sequences that encode fatty acid Δ12-epoxygenase enzymes as defined herein. More particularly, the present invention provides cDNA and genomic gene sequences that encode plant fatty acid epoxygenases, in particular from  Crepis palaestina  or  Vernonia galamensis . The genetic sequences of the present invention provide the means by which fatty acid metabolism may be altered or manipulated in organisms such as yeasts, moulds, bacteria, insects, birds, mammals and plants, in particular to convert unsaturated fatty acids to epoxy fatty acids therein. The invention extends to genetically modified oil-accumulating organisms transformed with the subject genetic sequences and to the oils derived therefrom. The oils thus produced provide the means for the cost-effective raw materials for use in the efficient production of coatings, resins, glues, plastics, surfactants and lubricants, amongst others.  
         GENERAL  
         [0003]    Those skilled in the art will be aware that the present invention is subject to variations and modifications other than those specifically described herein. It is to be understood that the invention includes all such variations and modifications. The invention also includes all such steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any two or more of said steps or features.  
           [0004]    Throughout this specification, unless the context requires otherwise, the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated integer or group of integers but not the exclusion of any other integer or group of integers.  
           [0005]    Bibliographic details of the publications referred to by author in this specification are collected at the end of the description.  
           [0006]    This specification contains nucleotide sequence information prepared using the program Patentin Version 3.1 presented herein after the claims. Each nucleotide sequence is identified in the sequence listing by the numeric indicator &lt;210&gt; followed by the sequence identifier [e.g. &lt;210&gt;1, &lt;210&gt;2, etc]. The length, type of sequence [DNA, protein (PRT), etc] and source organism for each nucleotide sequence are indicated by information provided in the numeric indicator fields &lt; 211 &gt;,&lt; 212 &gt; and &lt;213&gt;, respectively. Nucleotide sequences referred to in the specification are defined by the term “SEQ ID NO:”, followed by the sequence identifier [e.g. SEQ ID NO:1 refers to the sequence in the sequence listing designated as &lt;400&gt;1].  
         BACKGROUND TO THE INVENTION  
         [0007]    There is considerable interest world-wide in producing chemical feedstock, such as fatty acids, for industrial use from renewable plant sources rather than from non-renewable petrochemicals. This concept has broad appeal to manufacturers and consumers on the basis of resource conservation and provides a significant opportunity to develop new industrial crops for agriculture.  
           [0008]    There is a diverse array of unusual fatty acids in nature and these have been well characterized (Badam &amp; Patil, 1981; Smith, 1970). Many of these unusual fatty acids have industrial potential and this has led to interest in domesticating such species to enable agricultural production of particular fatty acids.  
           [0009]    One class of fatty acids of particular interest are the epoxy-fatty acids, consisting of an acyl chain in which two adjacent carbon bonds are linked by an epoxy bridge. Due to their high reactivity, they have considerable application in the production of coatings, resins, glues, plastics, surfactants and lubricants. These fatty acids are currently produced by chemical epoxidation of vegetable oils, mainly soybean oil and linseed oil, however this process produces mixtures of multiple and isomeric forms and involves significant processing costs.  
           [0010]    Attempts are being made by others to develop some wild plants that contain epoxy fatty acids (e.g.  Euphorbia lagascae , or  Vernonia galamensis ) into commercial sources of these oils. However, problems with agronomic suitability and low yield potential severely limit the commercial utility of traditional plant breeding and cultivation approaches.  
           [0011]    The rapidly increasing sophistication of recombinant DNA technology is greatly facilitating the efficiency of commercially-important industrial processes, by the expression of genes isolated from a first organism or species in a second organism or species to confer novel phenotypes thereon. More particularly, conventional industrial processes can be made more efficient or cost-effective, resulting in greater yields per unit cost by the application of recombinant DNA techniques.  
           [0012]    Moreover, the appropriate choice of host organism for the expression of a genetic sequence of interest provides for the production of compounds that are not normally produced or synthesized by the host, at a high yield and purity.  
           [0013]    However, despite the general effectiveness of recombinant DNA technology, the isolation of genetic sequences which encode important enzymes in fatty acid metabolism, in particular the genes which encode the fatty acid Δ12-epoxygenase enzymes responsible for producing 12,13-epoxy-9-octadecenoic acid (vernolic acid) and 12,13-epoxy-9,15-octadecadienoic acid, amongst others, remains a major obstacle to the development of genetically-engineered organisms which produce these fatty acids.  
           [0014]    Until the present invention, there were only limited biochemical data indicating the nature of fatty acid epoxygenase enzymes, in particular Δ12-epoxygenases. However, in  Euphorbia lagascae , the formation of 12,13-epoxy-9-octadecenoic acid (vernolic acid) from linoleic acid appears to be catalyzed by a cytochrome-P450-dependent Δ12 epoxygenase enzyme (Bafor et al., 1993; Blee et al., 1994). Additionally, developing seed of linseed plants have the capability to convert added vernolic acid to 12,13-epoxy-9,15-octadecadienoic acid by an endogenous Δ15 desaturase (Engeseth and Stymne, 1996). Epoxy-fatty acids can also be produced by a peroxide-dependent peroxygenase in plant tissues (Blee and Schuber, 1990).  
           [0015]    In work leading up to the present invention, the inventors sought to isolate genetic sequences which encode genes which are important for the production of epoxy-fatty acids, such as 12,13-epoxy-9-octadecenoic acid (vernolic acid) or 12,13-epoxy-9,15-octadecadienoic acid and to transfer these genetic sequences into highly productive commercial oilseed plants and/or other oil accumulating organisms.  
         SUMMARY OF THE INVENTION  
         [0016]    One aspect of the invention provides an isolated nucleic acid which encodes or is complementary to an isolated nucleic acid which encodes a fatty acid epoxygenase.  
           [0017]    A second aspect of the invention provides an isolated nucleic acid which hybridizes under at least low stringency conditions to at least 20 contiguous nucleotides of SEQ ID NOS:1 or 3 or 5 or 19 or 19, or a complementary sequence thereto.  
           [0018]    A further aspect of the invention provides isolated nucleic acid comprising a sequence of nucleotides selected from the group consisting of:  
           [0019]    (i) a nucleotide sequence that is at least 65% identical to a sequence selected from the group consisting of: SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 19;  
           [0020]    (ii) a nucleotide sequence that encodes an amino acid sequence that is at least about 50% identical to a sequence selected from the group consisting of: SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, and SEQ ID NO: 20; and  
           [0021]    (iii) a nucleotide sequence that is complementary to (i) or (ii).  
           [0022]    A further aspect of the invention provides a gene construct that comprises the isolated nucleic acid supra, in either the sense or antisense orientation, in operable connection with a promoter sequence.  
           [0023]    A further aspect of the invention provides a method of altering the level of epoxy fatty acids in a cell, tissue, organ or organism, said method comprising expressing a sense, antisense, ribozyme or co-suppression molecule comprising the isolated nucleic acid supra in said cell, tissue, organ or organism for a time and under conditions sufficient for the level of epoxy fatty acids therein to be increased or reduced.  
           [0024]    A further aspect of the invention provides a method of producing a recombinant enzymatically active epoxygenase polypeptide in a cell, said method comprising expressing the isolated nucleic acid supra in said cell for a time and under conditions sufficient for the epoxygenase encoded therefor to be produced.  
           [0025]    A further aspect of the invention provides a method of producing a recombinant enzymatically active epoxygenase polypeptide in a cell, said method comprising the steps of:  
           [0026]    (i) producing a gene construct which comprises the isolated nucleic acid supra placed operably under the control of a promoter capable of conferring expression on said genetic sequence in said cell, and optionally an expression enhancer element;  
           [0027]    (ii) transforming said gene construct into said cell; and  
           [0028]    (iii) selecting transformants which express a functional epoxygenase encoded by the genetic sequence at a high level.  
           [0029]    A still further aspect of the invention provides a method of producing a recombinant and enzymatically active epoxygenase polypeptide in a transgenic plant comprising the steps of:  
           [0030]    (i) producing a gene construct which comprises the isolated nucleic acid supra placed operably under the control of a seed-specific promoter and optionally an expression enhancer element, wherein said genetic sequences is also placed upstream of a transcription terminator sequence;  
           [0031]    (ii) transforming said gene construct into a cell or tissue of said plant; and  
           [0032]    (iii) selecting transformants which express a functional epoxygenase encoded by the genetic sequence at a high level in seeds.  
           [0033]    A further aspect of the invention provides a recombinant epoxygenase polypeptide or functional enzyme molecule.  
           [0034]    A further aspect of the invention provides a recombinant epoxygenase which comprises a sequence of amino acids set forth in any one of SEQ ID NOS: 2 or 4 or 6 or 20 or 20 or a homologue, analogue or derivative thereof which is at least about 50% identical thereto. More preferably, the percentage identity to any one of SEQ ID NOS: 2 or 4 or 6 or 20 or 20 is at least about 65%.  
           [0035]    A still further aspect of the invention provides a method of producing an epoxy fatty acid in a cell, tissue, organ or organism, said method comprising incubating a cell, tissue, organ or organism which expresses an enzymatically active recombinant epoxygenase with a fatty acid substrate and preferably, an unsaturated fatty acid substrate, for a time and under conditions sufficient for at least one carbon bond, preferably a carbon double bond, of said substrate to be converted to an epoxy group.  
           [0036]    A further aspect of the invention provides an immunologically interactive molecule which binds to the recombinant epoxygenase polypeptide described herein or a homologue, analogue or derivative thereof. 
       
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS  
       [0037]    [0037]FIG. 1 is a linear representation of an expression plasmid comprising an epoxygenase structural gene, placed operably under the control of the truncated napin promoter (FP1; right-hand hatched box) and placed upstream of the NOS terminator sequence (right-hand stippled box). The epoxygenase genetic sequence is indicated by the right-hand open rectangular box. The construct also comprises the NOS promoter (left-hand hatched box) driving expression of the NPTII 71 gene (left-hand open box) and placed upstream of the NOS terminator (left-hand stippled box). The left and right border sequences of the  Agrobacterium tumefaciens  Ti plasmid are also indicated.  
         [0038]    [0038]FIG. 2 is a schematic representation showing the alignment of the amino acid sequences of the epoxygenase polypeptide of  Crepis palaestina  (Cpa12; SEQ ID NO: 2), a further epoxygenase derived from Crepis sp. other than  C. palaestina  which produces high levels of vernolic acid (CrepX; SEQ ID NO: 4), a partial amino acid sequence of an epoxygenase polypeptide derived from  Vernonia galamensis  (Vgal1; SEQ ID NO: 6), a full-length amino acid sequence of an epoxygenase polypeptide derived from  Vernonia galamensis  (SEQ ID NO: 20), the amino acid sequence of the Δ12 acetylenase of  Crepis alpina  (Crep1; SEQ ID NO: 8), the Δ12 desaturase of  A. thaliana  (L26296; SEQ ID NO: 9),  Brassica juncea  (X91139; SEQ ID NO: 10),  Glycine max  (L43921; SEQ ID NO: 11),  Solanum commersonii  (X92847; SEQ ID NO: 12) and  Glycine max  (L43920; SEQ ID NO: 13), and the Δ12 hydroxylase of  Ricinus communis  (U22378; SEQ ID NO: 14). Underlined are three histidine-rich motifs that are conserved in non-heme containing mixed-function monooxygenases.  
         [0039]    [0039]FIG. 3 is a copy of a photographic representation of a northern blot hybridization showing seed-specific expression of the  Crepis palaestina  epoxygenase gene exemplified by SEQ ID NO: 1. Northern blot analysis of total RNA from leaves (lane 1) and developing seeds (lane 2) of  Crepis palaestina.  15 μg of total RNA was run on a Northern gel and blotted onto Hybond N +  membrane from Amersham according to the manufacturer&#39;s instructions. The blot was hybridized at 60° C. with a probe made from the 3′ untranslated region of SEQ ID NO: 1. The blot was washed twice in 2×SSC (NaCl-Sodium Citrate buffer) at room temperature for 10 minutes, then in 0.1×SSC at 60° C. for 20 min.  
         [0040]    [0040]FIG. 4 is a schematic representation of a binary plasmid vector containing an expression cassette comprising the truncated napin seed-specific promoter (Napin) and nopaline synthase terminator (NT), with a BamHI cloning site there between, in addition to the kanamycin-resistance gene NPII operably connected to the nopaline synthase promoter (NP) and nopaline synthase terminator (NT) sequences. The expression cassette is flanked by T-DNA left border (LB) and right-border (RB) sequences.  
         [0041]    [0041]FIG. 5 is a schematic representation of a binary plasmid vector containing an expression cassette which comprises SEQ ID NO: 1 placed operably under the control of a truncated napin seed-specific promoter (Napin) and upstream of the nopaline synthase terminator (NT), in addition to the kanamycin-resistance gene NPTII operably connected to the nopaline synthase promoter (NP) and nopaline synthase terminator (NT) sequences. The expression cassette is flanked by T-DNA left border (LB) and right-border (RB) sequences. To produce this construct, SEQ ID NO: 1 is inserted into the BamHI site of the binary vector set forth in FIG. 4.  
         [0042]    [0042]FIG. 6 is a graphical representation of gas-chromatography traces of fatty acid methyl esters prepared from oil seeds of untransformed  Arabidopsis thaliana  plants [panel (a)], or  A. thaliana  plants (transgenic line Cpal-17) which have been transformed with SEQ ID NO: 1 using the gene construct set forth in FIG. 5 [panels (b) and (c)]. In panels (a) and (b), fatty acid methyl esters were separated using packed column separation. In panel (c), the fatty acid methyl esters were separated using capillary column separation. The elution positions of vernolic acid are indicated.  
         [0043]    [0043]FIG. 7 is a graphical representation showing the joint distribution of epoxy fatty acids in selfed seed on T 1  plants of Cpal2-transformed  Arabidopsis thaliana  plants as determined using gas chromatography. Levels of both vernolic acid (x-axis) and 12,13-epoxy-9,15-octadecadienoic acid (y-axis) were determined and plotted relative to each other. Data show a positive correlation between the levels of these fatty acids in transgenic plants.  
         [0044]    [0044]FIG. 8 is a graphical representation showing the incorporation of  14 C-label into the chloroform phase obtained from lipid extraction of linseed cotyledons during labeled-substrate feeding. Symbols used; ♦, [ 14 C] oleic acid feeding; ▪, [ 14 C] vernolic acid feeding.  
         [0045]    [0045]FIG. 9 is a graphical representation showing the incorporation of  14 C-label into the phosphatidyl choline of linseed cotyledons during labeled-substrate feeding. Symbols used; ♦, [ 14 C] oleic acid feeding; ▪, [ 14 C] vernolic acid feeding.  
         [0046]    [0046]FIG. 10 is a graphical representation showing the incorporation of  14 C-label into the triacylglycerols of linseed cotyledons during labeled-substrate feeding. Symbols used♦, [ 14 C]oleic acid feeding; ▪, [ 14 C] vernolic acid feeding. 
     
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS  
       [0047]    One aspect of the present invention provides an isolated nucleic acid which encodes or is complementary to an isolated nucleic acid which encodes a fatty acid epoxygenase.  
         [0048]    Wherein the isolated nucleic acid of the invention encodes an enzyme which is involved in the direct epoxidation of arachidonic acid, it is particularly preferred that the subject nucleic acid is derived from a non-mammalian source.  
         [0049]    As used herein, the term “derived from” shall be taken to indicate that a particular integer or group of integers has originated from the species specified, but has not necessarily been obtained directly from the specified source.  
         [0050]    The term “non-mammalian source” refers to any organism other than a mammal or a tissue or cell derived from same. In the present context, the term “derived from a non-mammalian source” shall be taken to indicate that a particular integer or group of integers has been derived from bacteria, yeasts, birds, amphibians, reptiles, insects, plants, fungi, moulds and algae or other non-mammal.  
         [0051]    In a preferred embodiment of the present invention, the source organism is any such organism possessing the genetic capacity to synthesize epoxy fatty acids. More preferably, the source organism is a plant such as, but not limited to Chrysanthemum spp., Crepis spp., Euphorbia spp. and Vernonia spp., amongst others.  
         [0052]    Even more preferably, the source organism is selected from the group consisting of:  Crepis biennis, Crepis aurea, Crepis conyzaefolia, Crepis intermedia, Crepis occidentalis, Crepis palaestina, Crepis vesicaria, Crepis xacintha, Euphorbia lagascae  and  Vernonia galamensis . Additional species are not excluded.  
         [0053]    In a particularly preferred embodiment of the present invention, the source organism is a Crepis sp. comprising high levels of vernolic acid such as  Crepis palaestina , amongst others or alternatively,  Vernonia galamensis.    
         [0054]    Wherein the isolated nucleic acid of the invention encodes a Δ6-epoxygenase or Δ9-epoxygenase enzyme or Δ12-epoxygenase or Δ15-epoxygenase enzyme, or at least encodes an enzyme which is not involved in the direct epoxidation of arachidonic acid, the subject nucleic acid may be derived from any source producing said enzyme, including, but not limited to, yeasts, moulds, bacteria, insects, birds, mammals and plants.  
         [0055]    The nucleic acid of the invention according to any of the foregoing embodiments may be DNA, such as a gene, cDNA molecule, RNA molecule or a synthetic oligonucleotide molecule, whether single-stranded or double-stranded and irrespective of any secondary structure characteristics unless specifically stated.  
         [0056]    Reference herein to a “gene” is to be taken in its broadest context and includes:  
         [0057]    (i) a classical genomic gene consisting of transcriptional and/or translational regulatory sequences and/or a coding region and/or non-translated sequences (i.e. introns, 5′- and 3′-untranslated sequences);or  
         [0058]    (ii) mRNA or cDNA corresponding to the coding regions (i.e. exons) and 5′- and 3′- untranslated sequences of the gene.  
         [0059]    The term “gene” is also used to describe synthetic or fusion molecules encoding all or part of a functional product. Preferred epoxygenase genes of the present invention may be derived from a natural epoxygenase gene by standard recombinant techniques. Generally, an epoxygenase gene may be subjected to mutagenesis to produce single or multiple nucleotide substitutions, deletions and/or additions.  
         [0060]    Insertions are those variants in which one or more nucleotides are introduced into a predetermined site in the nucleotide sequence, although random insertion is also possible with suitable screening of the resulting product. Nucleotide insertions include 5′ and 3′ terminal fusions as well as intra-sequence insertions of single or multiple nucleotides.  
         [0061]    Deletions are variants characterized by the removal of one or more nucleotides from the sequence.  
         [0062]    Substitutions are those variants in which at least one nucleotide in the sequence has been removed and a different nucleotide inserted in its place. Such a substitution may be “silent” in that the substitution does not change the amino acid defined by the codon. Alternatively, a conservative substitution may alter one amino acid for another similar acting amino acid, or an amino acid of like charge, polarity, or hydrophobicity.  
         [0063]    In the context of the present invention, the term “fatty acid epoxygenase” shall be taken to refer to any enzyme or functional equivalent or enzymatically-active derivative thereof that catalyzes the biosynthesis of an epoxy fatty acid, by converting a carbon bond of a fatty acid to an epoxy group and preferably, by converting a carbon double bond of an unsaturated fatty acid to an epoxy group. Although not limiting the invention, a fatty acid epoxygenase may catalyze the biosynthesis of an epoxy fatty acid selected from the group consisting of: (i) 12,13-epoxy-9-octadecenoic acid (vernolic acid); (ii) 12,13-epoxy-9,15-octadecadienoic acid; (iii) 15,16-epoxy-9,12-octadecadienoic acid; (iv) 9,10-epoxy-12-octadecenoic acid; and (v) 9,10-epoxy-octadecanoic acid.  
         [0064]    The term “epoxy”, or “epoxy group” or “epoxy residue” will be known by those skilled in the art to refer to a three member ring comprising two carbon atoms and an oxygen atom linked by single bonds as follows:  
                         
 
         [0065]    Accordingly, the term “epoxide” refers to a compound that comprise at least one epoxy group as herein before defined.  
         [0066]    Those skilled in the art are aware that fatty acid nomenclature is based upon the length of the carbon chain and the position of unsaturated carbon atoms within that carbon chain. Thus, fatty acids are designated using the shorthand notation: 
         (Carbon) total (carbon double bonds) total   carbon double bond(Δ) position   
         [0067]    wherein the double bonds are cis unless otherwise indicated. For example, palmitic acid (n-hexadecanoic acid) is a saturated 16-carbon fatty acid (i.e. 16:0), oleic acid (octadecenoic acid) is an unsaturated 18-carbon fatty acid with one double bond between C-9 and C-10 (i.e. 18:1 Δ9 ), and linoleic acid (octadecadienoic acid) is an unsaturated 18-carbon fatty acid with two double bonds between C-9 and C-10 and between C-12 and C-13 (i.e. 18:2 Δ9,12 ).  
         [0068]    However, in the present context an epoxygenase enzyme may catalyze the conversion of any carbon bond to an epoxy group or alternatively, the conversion of any double in an unsaturated fatty acid substrate to an epoxy group. In this regard, it is well-known by those skilled in the art that most mono-unsaturated fatty acids of higher organisms are 18-carbon unsaturated fatty acids (i.e. 18:1 Δ9 ), while most polyunsaturated fatty acids derived from higher organisms are 18-carbon fatty acids with at least one of the double bonds therein located between C-9 and C-10. Additionally, bacteria also possess C16-mono-unsaturated fatty acids. Moreover, the epoxygenase of the present invention may act on more than a single fatty acid substrate molecule and, as a consequence, the present invention is not to be limited by the nature of the substrate molecule upon which the subject epoxygenase enzyme acts.  
         [0069]    Preferably, the substrate molecule for the epoxygenase of the present invention is an unsaturated fatty acid comprising at least one double bond.  
         [0070]    Furthermore, epoxygenase enzymes may act upon any number of carbon atoms in any one substrate molecule. For example, they may be characterized as Δ6-epoxygenase, Δ9-epoxygenase, Δ12-epoxygenase or Δ15-epoxygenase enzymes amongst others. Accordingly, the present invention is not limited by the position of the carbon atom in the substrate upon which an epoxygenase enzyme may act.  
         [0071]    The term “Δ6-epoxygenase” as used herein shall be taken to refer to an epoxygenase enzyme which catalyzes the conversion of the Δ6 carbon bond of a fatty acid substrate to a Δ6 epoxy group and preferably, catalyzes the conversion of the Δ6 double bond of at least one unsaturated fatty acid to a Δ6 epoxy group.  
         [0072]    The term “Δ9-epoxygenase” as used herein shall be taken to refer to an epoxygenase enzyme which catalyzes the conversion of the Δ9 carbon bond of a fatty acid substrate to a Δ9 epoxy group and preferably, catalyzes the conversion of the Δ9 double bond of at least one unsaturated fatty acid to a Δ9 epoxy group.  
         [0073]    As used herein, the term “Δ12-epoxygenase” shall be taken to refer to an epoxygenase enzyme which catalyzes the conversion of the Δ12 carbon bond of a fatty acid substrate to a Δ12 epoxy group and preferably, catalyzes the conversion of the Δ12 double bond of at least one unsaturated fatty acid to a Δ12 epoxy group.  
         [0074]    As used herein, the term “Δ15-epoxygenase” shall be taken to refer to an epoxygenase enzyme which catalyzes the conversion of the Δ15 carbon bond of a fatty acid substrate to a Δ15 epoxy group and preferably, catalyzes the conversion of the Δ15 double bond of at least one unsaturated fatty acid to a Δ15 epoxy group.  
         [0075]    The present invention clearly extends to genetic sequences which encode all of the epoxygenase enzymes listed supra, amongst others.  
         [0076]    In one preferred embodiment of the invention, the isolated nucleic acid encodes a fatty acid epoxygenase enzyme which converts at least one carbon bond in palmitoleic acid (16:1 Δ9 ), oleic acid (18:1 Δ9 ), linoleic acid (18:2 Δ9,12 ), linolenic acid (18:3 Δ9,12,15 ), or arachidoniic acid (20:4 Δ5,8,11,14 ) to an epoxy bond. Preferably, the carbon bond is a carbon double bond.  
         [0077]    More preferably, the isolated nucleic acid of the invention encodes a fatty acid epoxygenase enzyme that at least converts one or both double bonds in linoleic acid to an epoxy group. According to this embodiment, an epoxygenase which converts both the Δ9 and the Δ12 double bonds of linoleic acid to an epoxy group may catalyze such conversions independently of each other such that said epoxygenase is a Δ9-epoxygenase and/or a Δ12-epoxygenase enzyme as herein before defined.  
         [0078]    In an alternative preferred embodiment, the fatty acid epoxygenase of the present invention is a Δ12-epoxygenase, a Δ15- epoxygenase or a Δ9-epoxygenase as herein before defined.  
         [0079]    More preferably, the fatty acid epoxygenase of the invention is a Δ12- epoxygenase as herein before defined.  
         [0080]    In a particularly preferred embodiment of the invention, there is provided an isolated nucleic acid which encodes linoleate Δ12-epoxygenase, the enzyme which at least converts the Δ12 double bond of linoleic acid to a Δ12-epoxy group, thereby producing 12,13-epoxy-9-octadecenoic acid (vernolic acid).  
         [0081]    Although not limiting the present invention, the preferred source of the Δ12-epoxygenase of the invention is a plant, in particular  Crepis palaestina  or a further Crepis sp. which is distinct from  C. palaestina  but contains high levels of vernolic acid, or  Vernonia galamensis.    
         [0082]    According to this embodiment, a Δ12-epoxygenase may catalyze the conversion of palmitoleic acid to 9,10-epoxy-palmitic acid and/or the conversion of oleic acid to 9,10-epoxy-stearic acid and/or the conversion of linoleic acid to any one or more of 9,10-epoxy-12-octadecenoic acid or 12,13-epoxy-9-octadecenoic acid or 9,10,12,13-diepoxy-stearic acid and/or the conversion of linolenic acid to any one or more of 9,10-epoxy-12,15-octadecadienoic acid or 12,13-epoxy-9,15-octadecadienoic acid or 15,16-epoxy-octadecadienoic acid or 9,10,12,13-diepoxy-15-octadecenoic acid or 9,10,15,16-diepoxy-12-octadecenoic acid or 12,13,15,16-diepoxy-9-octadecenoic acid or 9,10,12,13,15,16-triepoxy-stearic acid and/or the conversion of arachidonic acid to any one or more of 5,6-epoxy-8,11,14-tetracosatrienoic acid or 8,9-epoxy-5,11,14-tetracosatrienoic acid or 11,12-epoxy-5,8,14-tetracosatrienoic acid or 14,15-epoxy-5,8,11 -tetracosatrienoic acid or 5,6,8,9-diepoxy-11,14-tetracosadienoic acid or 5,6,11,12-diepoxy-8,14-tetracosadienoic acid or 5,6,14,15-diepoxy-8,11-tetracosadienoic acid or 8,9,11,12-diepoxy-5,14-tetracosadienoic acid or 8,9,14,15-diepoxy-5,11-tetracosadienoic acid or 11,12,14,15-diepoxy-5,8-tetracosadienoic acid or 5,6,8,9,11,12-triepoxy-14-tetracosenoic acid or 5,6,8,9,14,15-triepoxy-11-tetracosenoic acid or 5,6,11,12,14,15-triepoxy-8-tetracosenoic acid or 8,9,11,12,14,15-triepoxy-5-tetracosenoic acid, amongst others.  
         [0083]    Those skilled in the art may be aware that not all substrates listed supra may be derivable from a natural source, but notwithstanding this, may be produced by chemical synthetic means. The conversion of both natural and synthetic unsaturated fatty acids to epoxy fatty acids is clearly within the scope of the present invention.  
         [0084]    The present invention is particularly directed to those epoxygenase enzymes that are mixed-function monooxygenase enzymes, and nucleic acids encoding said enzymes, and uses of said enzymes and nucleic acids. Accordingly, it is particularly preferred that the nucleic acid of the invention encode a fatty acid epoxygenase which is a mixed-function monooxygenase enzyme.  
         [0085]    In the context of the present invention, the term “mixed-function monooxygenase enzyme” shall be taken to refer to any epoxygenase polypeptide that comprises an amino acid sequence comprising three histidine-rich regions as follows: 
         [0086]    [0086]                               (i)   His-(Xaa) 3-4 -His (SEQ ID NO: 21 and SEQ ID NO: 22);                   (ii)   His-(Xaa) 2-3 -His-His (SEQ ID NO: 23 and SEQ ID NO: 24);               (iii)   His-(Xaa) 2-3 -His-His (SEQ ID NO: 23 and SEQ ID NO: 24),            
         [0087]    (i)  
         [0088]    (ii)  
         [0089]    (iii)  
         [0090]    and  
         [0091]    wherein His designates histidine, Xaa designates any naturally-occurring amino acid residue as set forth in Table 1 herein, the integer (Xaa) 3-4  refers to a sequence of amino acids comprising three or four repeats of Xaa, and the integer (Xaa) 2-3  refers to a sequence of amino acids comprising two or three repeats of Xaa.  
         [0092]    In the exemplification of the invention described herein, the inventors provide isolated cDNAs that comprise nucleotide sequences encoding the Δ12-epoxygenase polypeptides of  Crepis palaestina  and  Vernonia galamensis . Each exemplified full-length amino acid sequence encoded by said cDNAs which includes the three characteristic amino acid sequence motifs of a mixed-function monooxygenase enzyme as herein before defined. Close sequence identity between the amino acid sequences of the Δ12-epoxygenase enzymes from  C. palaestina  (SEQ ID NO: 2), an unidentified Crepis sp (SEQ ID NO: 4), and  Vernonia galamensis  (SEQ ID NO: 20), suggests functional similarity between these polypeptides. In contrast, the amino acid sequences of these epoxygenases have lower identity to the amino acid sequences of a fatty acid desaturase or a fatty acid hydroxylase.  
         [0093]    It is even more preferred that the epoxygenase of the present invention at least comprises a sequence of amino acids which comprises three histidine-rich regions as follows:  
                               (i)   His-Glu-Cys-Gly-His-His   (SEQ ID NO: 15);               (ii)   His-Arg-Asn-His-His   (SEQ ID NO: 16);                       and               (iii)   His-Val-Met-His-His   (SEQ ID NO: 17) or                   His-Val-Leu-His-His   (SEQ ID NO: 18),          
 
         [0094]    (i)  
         [0095]    (ii)  
         [0096]    (iii)  
         [0097]    wherein His designates histidine, Glu designates glutamate, Cys designates cysteine, Gly designates glycine, Arg designates arginine, Asn designates asparagine, Val designates valine, Met designates methionine and Leu designates leucine.  
         [0098]    The present invention clearly extends to epoxygenase genes derived from other species, including the epoxygenase genes derived from Chrysanthemum spp. and  Euphorbia lagascae , amongst others.  
         [0099]    In a preferred embodiment, whilst not limiting the present invention, the epoxygenase genes of other species which are encompassed by the present invention encode mixed-function monooxygenase enzymes. The present invention further extends to the isolated or recombinant polypeptides encoded by such genes and uses of said genes and polypeptides.  
         [0100]    The invention described according to this embodiment does not encompass nucleic acids which encode enzyme activities other than epoxygenase activities as defined herein, in particular the Δ12-desaturase enzymes derived from  Arabidopsis thaliana, Brassica juncea, Brassica napus  or  Glycine max , amongst others, which are known to contain similar histidine-rich motifs.  
         [0101]    In the present context, “homologues” of an amino acid sequence refer to those amino acid sequences or peptide sequences which are derived from polypeptides, enzymes or proteins of the present invention or alternatively, correspond substantially to the amino acid sequences listed supra, notwithstanding any naturally-occurring amino acid substitutions, additions or deletions thereto.  
         [0102]    For example, amino acids may be replaced by other amino acids having similar properties, for example hydrophobicity, hydrophilicity, hydrophobic moment, antigenicity, propensity to form or break α-helical structures or β-sheet structures, and so on. Alternatively, or in addition, the amino acids of a homologous amino acid sequence may be replaced by other amino acids having similar properties, for example hydrophobicity, hydrophilicity, hydrophobic moment, charge or antigenicity, and so on.  
         [0103]    Naturally-occurring amino acid residues contemplated herein are described in Table 1.  
         [0104]    A homologue of an amino acid sequence may be a synthetic peptide produced by any method known to those skilled in the art, such as by using Fmoc chemistry.  
         [0105]    Alternatively, a homologue of an amino acid sequence may be derived from a natural source, such as the same or another species as the polypeptides, enzymes or proteins of the present invention. Preferred sources of homologues of the amino acid sequences listed supra include any of the sources contemplated herein.  
         [0106]    “Analogues” of an amino acid sequence encompass those amino acid sequences which are substantially identical to the amino acid sequences listed supra notwithstanding the occurrence of any non-naturally occurring amino acid analogues therein.  
         [0107]    Preferred non-naturally occurring amino acids contemplated herein are listed below in Table 2.  
         [0108]    The term “derivative” in relation to an amino acid sequence shall be taken to refer hereinafter to mutants, parts, fragments or polypeptide fusions of the amino acid sequences listed supra. Derivatives include modified amino acid sequences or peptides in which ligands are attached to one or more of the amino acid residues contained therein, such as carbohydrates, enzymes, proteins, polypeptides or reporter molecules such as radionuclides or fluorescent compounds. Glycosylated, fluorescent, acylated or alkylated forms of the subject peptides are also contemplated by the present invention. Additionally, derivatives may comprise fragments or parts of an amino acid sequence disclosed herein and are within the scope of the invention, as are homopolymers or heteropolymers comprising two or more copies of the subject sequences.  
         [0109]    Procedures for derivatizing peptides are well-known in the art.  
         [0110]    Substitutions encompass amino acid alterations in which an amino acid is replaced with a different naturally-occurring or a non-conventional amino acid residue. Such substitutions may be classified as “conservative”, in which case an amino acid residue is replaced with another naturally-occurring amino acid of similar character, for example Gly⇄Ala, Val⇄Ile⇄Leu, Asp⇄Glu, Lys⇄Arg, Asn⇄Gln or Phe⇄Trp⇄Tyr.  
         [0111]    Substitutions encompassed by the present invention may also be “non-conservative”, in which an amino acid residue which is present in a repressor polypeptide is substituted with an amino acid having different properties, such as a naturally-occurring amino acid from a different group (e.g. substituted a charged or hydrophobic amino acid with alanine), or alternatively, in which a naturally-occurring amino acid is substituted with a non-conventional amino acid.  
         [0112]    Amino acid substitutions are typically of single residues, but may be of multiple residues, either clustered or dispersed.  
         [0113]    Amino acid deletions will usually be of the order of about 1-10 amino acid residues, while insertions may be of any length. Deletions and insertions may be made to the N-terminus, the C-terminus or be internal deletions or insertions. Generally, insertions within the amino acid sequence will be smaller than amino-or carboxyl-terminal fusions and of the order of 1-4 amino acid residues.  
         [0114]    The present invention clearly extends to the subject isolated nucleic acid when integrated into the genome of a cell as an addition to the endogenous cellular complement of epoxygenase genes. Alternatively, wherein the host cell does not normally encode enzymes required for epoxy fatty acid biosynthesis, the present invention extends to the subject isolated nucleic acid when integrated into the genome of said cell as an addition to the endogenous cellular genome.  
                               TABLE 1                                       Three-letter   One-letter           Amino Acid   Abbreviation   Symbol                           Alanine   Ala   A           Arginine   Arg   R           Asparagine   Asn   N           Aspartic acid   Asp   D           Cysteine   Cys   C           Glutamine   Gln   Q           Glutamic acid   Glu   E           Glycine   Gly   G           Histidine   His   H           Isoleucine   Ile   I           Leucine   Leu   L           Lysine   Lys   K           Methionine   Met   M           Phenylalanine   Phe   F           Proline   Pro   P           Serine   Ser   S           Threonine   Thr   T           Tryptophan   Trp   W           Tyrosine   Tyr   Y           Valine   Val   V           Any amino acid as above   Xaa   X                      
 
         [0115]    [0115]                           TABLE 2                                   Non-conventional               amino acid   Code                           α-aminobutyric acid   Abu           α-amino-α-methylbutyrate   Mgabu           aminocyclopropane-   Cpro           carboxylate               aminoisobutyric acid   Aib           aminonorbornyl-   Norb           carboxylate               cyclohexylalanine   Chexa           cyclopentylalanine   Cpen           D-alanine   Dal           D-arginine   Darg           D-aspartic acid   Dasp           D-cysteine   Dcys           D-glutamine   Dgln           D-glutamic acid   Dglu           D-histidine   Dhis           D-isoleucine   Dile           D-leucine   Dleu           D-lysine   Dlys           D-methionine   Dmet           D-ornithine   Dorn           D-phenylalanine   Dphe           D-proline   Dpro           D-serine   Dser           D-threonine   Dthr           D-tryptophan   Dtrp           D-tyrosine   Dtyr           D-valine   Dval           D-α-methylalanine   Dmala           D-α-methylarginine   Dmarg           D-α-methylasparagine   Dmasn           D-α-methylaspartate   Dmasp           D-α-methylcysteine   Dmcys           D-α-methylglutamine   Dmgln           D-α-methylhistidine   Dmhis           D-α-methylisoleucine   Dmile           D-α-methylleucine   Dmleu           D-α-methyllysine   Dmlys           D-α-methylmethionine   Dmmet           D-α-methylornithine   Dmorn           D-α-methylphenylalanine   Dmphe           D-α-methylproline   Dmpro           D-α-methylserine   Dmser           D-α-methylthreonine   Dmthr           D-α-methyltryptophan   Dmtrp           D-α-methyltyrosine   Dmty           D-α-methylvaline   Dmval           D-N-methylalanine   Dnmala           D-N-methylarginine   Dnmarg           D-N-methylasparagine   Dnmasn           D-N-methylaspartate   Dnmasp           D-N-methylcysteine   Dnmcys           D-N-methylglutamine   Dnmgln           D-N-methylglutamate   Dnmglu           D-N-methylhistidine   Dnmhis           D-N-methylisoleucine   Dnmile           D-N-methylleucine   Dnmleu           D-N-methyllysine   Dnmlys           N-methylcyclohexylalanine   Nmchexa           D-N-methylornithine   Dnmorn           L-N-methylalanine   Nmala           L-N-methylarginine   Nmarg           L-N-methylasparagine   Nmasn           L-N-methylaspartic acid   Nmasp           L-N-methylcysteine   Nmcys           L-N-methylglutamine   Nmgln           L-N-methylglutamic acid   Nmglu           L-N-methylhistidine   Nmhis           L-N-methylisolleucine   Nmile           L-N-methylleucine   Nmleu           L-N-methyllysine   Nmlys           L-N-methylmethionine   Nmmet           L-N-methylnorleucine   Nmnle           L-N-methylnorvaline   Nmnva           L-N-methylornithine   Nmorn           L-N-methylphenylalanine   Nmphe           L-N-methylproline   Nmpro           L-N-methylserine   Nmser           L-N-methylthreonine   Nmthr           L-N-methyltryptophan   Nmtrp           L-N-methyltyrosine   Nmtyr           L-N-methylvaline   Nmval           L-N-methylethylglycine   Nmetg           L-N-methyl-t-butylglycine   Nmtbug           L-norleucine   Nle           L-norvaline   Nva           α-methyl-aminoisobutyrate   Maib           α-methyl-γ-aminobutyrate   Mgabu           α-methylcyclohexylalanine   Mchexa           α-methylcylcopentylalanine   Mcpen           α-methyl-α-napthylalanine   Manap           α-methylpenicillamine   Mpen           N-(4-aminobutyl)glycine   Nglu           N-(2-aminoethyl)glycine   Naeg           N-(3-aminopropyl)glycine   Norn           N-amino-α-methylbutyrate   Nmaabu           α-napthylalanine   Anap           N-benzylglycine   Nphe           N-(2-carbamylethyl)glycine   Ngln           N-(carbamylmethyl)glycine   Nasn           N-(2-carboxyethyl)glycine   Nglu           N-(carboxymethyl)glycine   Nasp           N-cyclobutylglycine   Ncbut           N-cycloheptylglycine   Nchep           N-cyclohexylglycine   Nchex           N-cyclodecylglycine   Ncdec           N-cylcododecylglycine   Ncdod           N-cyclooctylglycine   Ncoct           N-cyclopropylglycine   Ncpro           N-cycloundecylglycine   Ncund           N-(2,2-diphenylethyl)   Nbhm           glycine           N-(3,3-diphenylpropyl)   Nbhe           glycine           N-(3-guanidinopropyl)   Narg           glycine           N-(1-hydroxyethyl)glycine   Nthr           N-(hydroxyethyl))glycine   Nser           N-(imidazolylethyl))   Nhis           glycine           N-(3-indolylyethyl)   Nhtrp           glycine           N-methyl-γ-aminobutyrate   Nmgabu           D-N-methylmethionine   Dnmmet           N-methylcyclopentylalanine   Nmcpen                        
         [0116]    [0116] 
     
       
       
         1 
         
           
             24  
           
           
             1  
             1358  
             DNA  
             Crepis palaestina  
             
               CDS  
               (30)..(1151)  
                 
             
           
            1 

gagaagttga ccataaatca tttatcaac atg ggt gcc ggc ggt cgt ggt cgg       53 
                                Met Gly Ala Gly Gly Arg Gly Arg 
                                1               5 

aca tcg gaa aaa tcg gtc atg gaa cgt gtc tca gtt gat cca gta acc      101 
Thr Ser Glu Lys Ser Val Met Glu Arg Val Ser Val Asp Pro Val Thr 
    10                  15                  20 

ttc tca ctg agt gaa ttg aag caa gca atc cct ccc cat tgc ttc cag      149 
Phe Ser Leu Ser Glu Leu Lys Gln Ala Ile Pro Pro His Cys Phe Gln 
25                  30                  35                  40 

aga tct gta atc cgc tca tct tac tat gtt gtt caa gat ctc att att      197 
Arg Ser Val Ile Arg Ser Ser Tyr Tyr Val Val Gln Asp Leu Ile Ile 
                45                  50                  55 

gcc tac atc ttc tac ttc ctt gcc aac aca tat atc cct act ctt cct      245 
Ala Tyr Ile Phe Tyr Phe Leu Ala Asn Thr Tyr Ile Pro Thr Leu Pro 
            60                  65                  70 

act agt cta gcc tac tta gct tgg ccc gtt tac tgg ttc tgt caa gct      293 
Thr Ser Leu Ala Tyr Leu Ala Trp Pro Val Tyr Trp Phe Cys Gln Ala 
        75                  80                  85 

agc gtc ctc act ggc tta tgg atc ctc ggc cac gaa tgt ggt cac cat      341 
Ser Val Leu Thr Gly Leu Trp Ile Leu Gly His Glu Cys Gly His His 
    90                  95                  100 

gcc ttt agc aac tac aca tgg ttt gac gac act gtg ggc ttc atc ctc      389 
Ala Phe Ser Asn Tyr Thr Trp Phe Asp Asp Thr Val Gly Phe Ile Leu 
105                 110                 115                 120 

cac tca ttt ctc ctc acc ccg tat ttc tct tgg aaa ttc agt cac cgg      437 
His Ser Phe Leu Leu Thr Pro Tyr Phe Ser Trp Lys Phe Ser His Arg 
                125                 130                 135 

aat cac cat tcc aac aca agt tcg att gat aac gat gaa gtt tac att      485 
Asn His His Ser Asn Thr Ser Ser Ile Asp Asn Asp Glu Val Tyr Ile 
            140                 145                 150 

ccg aaa agc aag tcc aaa ctc gcg cgt atc tat aaa ctt ctt aac aac      533 
Pro Lys Ser Lys Ser Lys Leu Ala Arg Ile Tyr Lys Leu Leu Asn Asn 
        155                 160                 165 

cca cct ggt cgg ctg ttg gtt ttg att atc atg ttc acc cta gga ttt      581 
Pro Pro Gly Arg Leu Leu Val Leu Ile Ile Met Phe Thr Leu Gly Phe 
    170                 175                 180 

cct tta tac ctc ttg aca aat att tcc ggc aag aaa tac gac agg ttt      629 
Pro Leu Tyr Leu Leu Thr Asn Ile Ser Gly Lys Lys Tyr Asp Arg Phe 
185                 190                 195                 200 

gcc aac cac ttc gac ccc atg agt cca att ttc aaa gaa cgt gag cgg      677 
Ala Asn His Phe Asp Pro Met Ser Pro Ile Phe Lys Glu Arg Glu Arg 
                205                 210                 215 

ttt cag gtc ttc ctt tcg gat ctt ggt ctt ctt gcc gtg ttt tat gga      725 
Phe Gln Val Phe Leu Ser Asp Leu Gly Leu Leu Ala Val Phe Tyr Gly 
            220                 225                 230 

att aaa gtt gct gta gca aat aaa gga gct gct tgg gta gcg tgc atg      773 
Ile Lys Val Ala Val Ala Asn Lys Gly Ala Ala Trp Val Ala Cys Met 
        235                 240                 245 

tat gga gtt ccg gta tta ggc gta ttt acc ttt ttc gat gtg atc acc      821 
Tyr Gly Val Pro Val Leu Gly Val Phe Thr Phe Phe Asp Val Ile Thr 
    250                 255                 260 

ttc ttg cac cac acc cat cag tcg tcg cct cat tat gat tca act gaa      869 
Phe Leu His His Thr His Gln Ser Ser Pro His Tyr Asp Ser Thr Glu 
265                 270                 275                 280 

tgg aac tgg atc aga ggg gcc ttg tca gca atc gat agg gac ttt gga      917 
Trp Asn Trp Ile Arg Gly Ala Leu Ser Ala Ile Asp Arg Asp Phe Gly 
                285                 290                 295 

ttc ctg aat agt gtt ttc cat gat gtt aca cac act cat gtc atg cat      965 
Phe Leu Asn Ser Val Phe His Asp Val Thr His Thr His Val Met His 
            300                 305                 310 

cat ttg ttt tca tac att cca cac tat cat gca aag gag gca agg gat     1013 
His Leu Phe Ser Tyr Ile Pro His Tyr His Ala Lys Glu Ala Arg Asp 
        315                 320                 325 

gca atc aag cca atc ttg ggc gac ttt tat atg atc gac agg act cca     1061 
Ala Ile Lys Pro Ile Leu Gly Asp Phe Tyr Met Ile Asp Arg Thr Pro 
    330                 335                 340 

att tta aaa gca atg tgg aga gag ggc agg gag tgc atg tac atc gag     1109 
Ile Leu Lys Ala Met Trp Arg Glu Gly Arg Glu Cys Met Tyr Ile Glu 
345                 350                 355                 360 

cct gat agc aag ctc aaa ggt gtt tat tgg tat cat aaa ttg             1151 
Pro Asp Ser Lys Leu Lys Gly Val Tyr Trp Tyr His Lys Leu 
                365                 370 

tgatcatatg caaaatgcac atgcattttc aaaccctcta gttacgtttg ttctatgtat   1211 

aataaaccgc cggtcctttg gttgactatg cctaagccag gcgaaacagt taaataatat   1271 

cggtatgatg tgtaatgaaa gtatgtggtt gtctggtttt gttgctatga aagaaagtat   1331 

gtggttgtcg gtcaaaaaaa aaaaaaa                                       1358 

 
           
             2  
             374  
             PRT  
             Crepis palaestina  
           
            2 

Met Gly Ala Gly Gly Arg Gly Arg Thr Ser Glu Lys Ser Val Met Glu 
1               5                   10                  15 

Arg Val Ser Val Asp Pro Val Thr Phe Ser Leu Ser Glu Leu Lys Gln 
            20                  25                  30 

Ala Ile Pro Pro His Cys Phe Gln Arg Ser Val Ile Arg Ser Ser Tyr 
        35                  40                  45 

Tyr Val Val Gln Asp Leu Ile Ile Ala Tyr Ile Phe Tyr Phe Leu Ala 
    50                  55                  60 

Asn Thr Tyr Ile Pro Thr Leu Pro Thr Ser Leu Ala Tyr Leu Ala Trp 
65                  70                  75                  80 

Pro Val Tyr Trp Phe Cys Gln Ala Ser Val Leu Thr Gly Leu Trp Ile 
                85                  90                  95 

Leu Gly His Glu Cys Gly His His Ala Phe Ser Asn Tyr Thr Trp Phe 
            100                 105                 110 

Asp Asp Thr Val Gly Phe Ile Leu His Ser Phe Leu Leu Thr Pro Tyr 
        115                 120                 125 

Phe Ser Trp Lys Phe Ser His Arg Asn His His Ser Asn Thr Ser Ser 
    130                 135                 140 

Ile Asp Asn Asp Glu Val Tyr Ile Pro Lys Ser Lys Ser Lys Leu Ala 
145                 150                 155                 160 

Arg Ile Tyr Lys Leu Leu Asn Asn Pro Pro Gly Arg Leu Leu Val Leu 
                165                 170                 175 

Ile Ile Met Phe Thr Leu Gly Phe Pro Leu Tyr Leu Leu Thr Asn Ile 
            180                 185                 190 

Ser Gly Lys Lys Tyr Asp Arg Phe Ala Asn His Phe Asp Pro Met Ser 
        195                 200                 205 

Pro Ile Phe Lys Glu Arg Glu Arg Phe Gln Val Phe Leu Ser Asp Leu 
    210                 215                 220 

Gly Leu Leu Ala Val Phe Tyr Gly Ile Lys Val Ala Val Ala Asn Lys 
225                 230                 235                 240 

Gly Ala Ala Trp Val Ala Cys Met Tyr Gly Val Pro Val Leu Gly Val 
                245                 250                 255 

Phe Thr Phe Phe Asp Val Ile Thr Phe Leu His His Thr His Gln Ser 
            260                 265                 270 

Ser Pro His Tyr Asp Ser Thr Glu Trp Asn Trp Ile Arg Gly Ala Leu 
        275                 280                 285 

Ser Ala Ile Asp Arg Asp Phe Gly Phe Leu Asn Ser Val Phe His Asp 
    290                 295                 300 

Val Thr His Thr His Val Met His His Leu Phe Ser Tyr Ile Pro His 
305                 310                 315                 320 

Tyr His Ala Lys Glu Ala Arg Asp Ala Ile Lys Pro Ile Leu Gly Asp 
                325                 330                 335 

Phe Tyr Met Ile Asp Arg Thr Pro Ile Leu Lys Ala Met Trp Arg Glu 
            340                 345                 350 

Gly Arg Glu Cys Met Tyr Ile Glu Pro Asp Ser Lys Leu Lys Gly Val 
        355                 360                 365 

Tyr Trp Tyr His Lys Leu 
    370 

 
           
             3  
             1309  
             DNA  
             Crepis sp.  
             
               misc_feature  
               (937)..(937)  
               N is any nucleotide residue  
             
           
            3 

tgttgaccat aaatcatcta tcaac atg ggt gcc ggc ggc cgt ggt cgg tcg       52 
                            Met Gly Ala Gly Gly Arg Gly Arg Ser 
                            1               5 

gaa aag tcg gtc atg gaa cgt gtc tca gtt gat cca gta acc ttc tca      100 
Glu Lys Ser Val Met Glu Arg Val Ser Val Asp Pro Val Thr Phe Ser 
10                  15                  20                  25 

ctg agt gat ttg aag caa gca atc cct cca cat tgc ttc cag cga tct      148 
Leu Ser Asp Leu Lys Gln Ala Ile Pro Pro His Cys Phe Gln Arg Ser 
                30                  35                  40 

gtc atc cgt tca tct tat tac gtt gtt cag gat ctc ata att gcc tac      196 
Val Ile Arg Ser Ser Tyr Tyr Val Val Gln Asp Leu Ile Ile Ala Tyr 
            45                  50                  55 

atc ttc tac ttc ctt gcc aac aca tat atc cct aat ctc cct cat cct      244 
Ile Phe Tyr Phe Leu Ala Asn Thr Tyr Ile Pro Asn Leu Pro His Pro 
        60                  65                  70 

cta gcc tac tta gct tgg ccg ctt tac tgg ttc tgt caa gct agc gtc      292 
Leu Ala Tyr Leu Ala Trp Pro Leu Tyr Trp Phe Cys Gln Ala Ser Val 
    75                  80                  85 

ctc act ggg tta tgg atc ctc ggc cat gaa tgt ggt cac cat gcc tat      340 
Leu Thr Gly Leu Trp Ile Leu Gly His Glu Cys Gly His His Ala Tyr 
90                  95                  100                 105 

agc aac tac aca tgg gtt gac gac act gtg ggc ttc atc atc cat tca      388 
Ser Asn Tyr Thr Trp Val Asp Asp Thr Val Gly Phe Ile Ile His Ser 
                110                 115                 120 

ttt ctc ctc acc ccg tat ttc tct tgg aaa tac agt cac cgg aat cac      436 
Phe Leu Leu Thr Pro Tyr Phe Ser Trp Lys Tyr Ser His Arg Asn His 
            125                 130                 135 

cat tcc aac aca agt tcg att gat aac gat gaa gtt tac att ccg aaa      484 
His Ser Asn Thr Ser Ser Ile Asp Asn Asp Glu Val Tyr Ile Pro Lys 
        140                 145                 150 

agc aag tcc aaa ctc aag cgt atc tat aaa ctt ctt aac aac cca cct      532 
Ser Lys Ser Lys Leu Lys Arg Ile Tyr Lys Leu Leu Asn Asn Pro Pro 
    155                 160                 165 

ggt cga ctg ttg gtt ttg gtt atc atg ttc acc cta gga ttt cct tta      580 
Gly Arg Leu Leu Val Leu Val Ile Met Phe Thr Leu Gly Phe Pro Leu 
170                 175                 180                 185 

tac ctc ttg aca aat att tcc ggc aag aaa tac gat agg ttt gcc aac      628 
Tyr Leu Leu Thr Asn Ile Ser Gly Lys Lys Tyr Asp Arg Phe Ala Asn 
                190                 195                 200 

cac ttc gac ccc atg agt cca att ttc aaa gaa cgt gag cgg ttt cag      676 
His Phe Asp Pro Met Ser Pro Ile Phe Lys Glu Arg Glu Arg Phe Gln 
            205                 210                 215 

gtc ttc ctt tcg gat ctt ggt ctt ctt gct gtg ttt tat gga att aaa      724 
Val Phe Leu Ser Asp Leu Gly Leu Leu Ala Val Phe Tyr Gly Ile Lys 
        220                 225                 230 

gtt gct gta gca aat aaa gga gct gct tgg gtg gcg tgc atg tat gga      772 
Val Ala Val Ala Asn Lys Gly Ala Ala Trp Val Ala Cys Met Tyr Gly 
    235                 240                 245 

gtt ccg gtg cta ggc gta ttt acc ttt ttc gat gtg atc acg ttc tta      820 
Val Pro Val Leu Gly Val Phe Thr Phe Phe Asp Val Ile Thr Phe Leu 
250                 255                 260                 265 

cac cac acc cat cag tcg tcg cct cat tat gat tca act gaa tgg aac      868 
His His Thr His Gln Ser Ser Pro His Tyr Asp Ser Thr Glu Trp Asn 
                270                 275                 280 

tgg atc aga ggg gct ttg tca gca atc gat agn gac ttt ggg ttc ctg      916 
Trp Ile Arg Gly Ala Leu Ser Ala Ile Asp Xaa Asp Phe Gly Phe Leu 
            285                 290                 295 

aat agt gtt ttc cat gat gtn aca cac act cac gtc atg cat cat ttg      964 
Asn Ser Val Phe His Asp Val Thr His Thr His Val Met His His Leu 
        300                 305                 310 

ttt tca tac att cca cac tat cat gca aag gaa gca agg gat gca atc     1012 
Phe Ser Tyr Ile Pro His Tyr His Ala Lys Glu Ala Arg Asp Ala Ile 
    315                 320                 325 

aaa ccg atc ttg ggc gac ttt tat atg atc gat agg act cca att tta     1060 
Lys Pro Ile Leu Gly Asp Phe Tyr Met Ile Asp Arg Thr Pro Ile Leu 
330                 335                 340                 345 

aaa gca atg tgg aga gag ggc agg gaa tgc atg tac atc gag cct gat     1108 
Lys Ala Met Trp Arg Glu Gly Arg Glu Cys Met Tyr Ile Glu Pro Asp 
                350                 355                 360 

agc aag ctc aaa ggt gtt tat tgg tat cat aaa ttg tga tcatatgcaa      1157 
Ser Lys Leu Lys Gly Val Tyr Trp Tyr His Lys Leu 
            365                 370 

aatgcacatg cattttcaaa ccctctagtt acctttgttc tatgtataat aagaccgccg   1217 

gtcctatggt tttctatgcc taagccaggc gaaatagtta aataatatcg gtatgatgta   1277 

atgaaagtat gtggttgtct aaaaaaaaaa aa                                 1309 

 
           
             4  
             373  
             PRT  
             Crepis sp.  
             
               misc_feature  
               (292)..(292)  
               The ′Xaa′ at location 292 stands for Arg, 
      or Ser.  
             
           
            4 

Met Gly Ala Gly Gly Arg Gly Arg Ser Glu Lys Ser Val Met Glu Arg 
1               5                   10                  15 

Val Ser Val Asp Pro Val Thr Phe Ser Leu Ser Asp Leu Lys Gln Ala 
            20                  25                  30 

Ile Pro Pro His Cys Phe Gln Arg Ser Val Ile Arg Ser Ser Tyr Tyr 
        35                  40                  45 

Val Val Gln Asp Leu Ile Ile Ala Tyr Ile Phe Tyr Phe Leu Ala Asn 
    50                  55                  60 

Thr Tyr Ile Pro Asn Leu Pro His Pro Leu Ala Tyr Leu Ala Trp Pro 
65                  70                  75                  80 

Leu Tyr Trp Phe Cys Gln Ala Ser Val Leu Thr Gly Leu Trp Ile Leu 
                85                  90                  95 

Gly His Glu Cys Gly His His Ala Tyr Ser Asn Tyr Thr Trp Val Asp 
            100                 105                 110 

Asp Thr Val Gly Phe Ile Ile His Ser Phe Leu Leu Thr Pro Tyr Phe 
        115                 120                 125 

Ser Trp Lys Tyr Ser His Arg Asn His His Ser Asn Thr Ser Ser Ile 
    130                 135                 140 

Asp Asn Asp Glu Val Tyr Ile Pro Lys Ser Lys Ser Lys Leu Lys Arg 
145                 150                 155                 160 

Ile Tyr Lys Leu Leu Asn Asn Pro Pro Gly Arg Leu Leu Val Leu Val 
                165                 170                 175 

Ile Met Phe Thr Leu Gly Phe Pro Leu Tyr Leu Leu Thr Asn Ile Ser 
            180                 185                 190 

Gly Lys Lys Tyr Asp Arg Phe Ala Asn His Phe Asp Pro Met Ser Pro 
        195                 200                 205 

Ile Phe Lys Glu Arg Glu Arg Phe Gln Val Phe Leu Ser Asp Leu Gly 
    210                 215                 220 

Leu Leu Ala Val Phe Tyr Gly Ile Lys Val Ala Val Ala Asn Lys Gly 
225                 230                 235                 240 

Ala Ala Trp Val Ala Cys Met Tyr Gly Val Pro Val Leu Gly Val Phe 
                245                 250                 255 

Thr Phe Phe Asp Val Ile Thr Phe Leu His His Thr His Gln Ser Ser 
            260                 265                 270 

Pro His Tyr Asp Ser Thr Glu Trp Asn Trp Ile Arg Gly Ala Leu Ser 
        275                 280                 285 

Ala Ile Asp Xaa Asp Phe Gly Phe Leu Asn Ser Val Phe His Asp Val 
    290                 295                 300 

Thr His Thr His Val Met His His Leu Phe Ser Tyr Ile Pro His Tyr 
305                 310                 315                 320 

His Ala Lys Glu Ala Arg Asp Ala Ile Lys Pro Ile Leu Gly Asp Phe 
                325                 330                 335 

Tyr Met Ile Asp Arg Thr Pro Ile Leu Lys Ala Met Trp Arg Glu Gly 
            340                 345                 350 

Arg Glu Cys Met Tyr Ile Glu Pro Asp Ser Lys Leu Lys Gly Val Tyr 
        355                 360                 365 

Trp Tyr His Lys Leu 
    370 

 
           
             5  
             550  
             DNA  
             Vernonia galamensis  
             
               CDS  
               (1)..(549)  
                 
             
           
            5 

cat cac gcc ttc agt gac tat caa tgg ata gac gac act gtg ggc ttc       48 
His His Ala Phe Ser Asp Tyr Gln Trp Ile Asp Asp Thr Val Gly Phe 
1               5                   10                  15 

atc ctt cac ttt gca ctc ttc acc cct tat ttc tct tgg aaa tac agt       96 
Ile Leu His Phe Ala Leu Phe Thr Pro Tyr Phe Ser Trp Lys Tyr Ser 
            20                  25                  30 

cac cgt aat cac cat gcc aac aca aac tct ctt gta acc gat gaa gta      144 
His Arg Asn His His Ala Asn Thr Asn Ser Leu Val Thr Asp Glu Val 
        35                  40                  45 

tac atc cct aaa gtt aaa tcc aag gtc aag att tat tcc aaa atc ctt      192 
Tyr Ile Pro Lys Val Lys Ser Lys Val Lys Ile Tyr Ser Lys Ile Leu 
    50                  55                  60 

aac aac cct cct ggt cgc gtt ttc acc ttg gct ttc aga ttg atc gtg      240 
Asn Asn Pro Pro Gly Arg Val Phe Thr Leu Ala Phe Arg Leu Ile Val 
65                  70                  75                  80 

ggt ttt cct tta tac ctt ttc acc aat gtt tca ggc aag aaa tac gaa      288 
Gly Phe Pro Leu Tyr Leu Phe Thr Asn Val Ser Gly Lys Lys Tyr Glu 
                85                  90                  95 

cgt ttt gcc aac cat ttt gat ccc atg agt ccc att ttc acc gag cgt      336 
Arg Phe Ala Asn His Phe Asp Pro Met Ser Pro Ile Phe Thr Glu Arg 
            100                 105                 110 

gag cat gta caa gtc ttg ctt tct gat ttt ggt ctc ata gca gtt gct      384 
Glu His Val Gln Val Leu Leu Ser Asp Phe Gly Leu Ile Ala Val Ala 
        115                 120                 125 

tac gtg gtt cgt caa gct gta ctg gct aaa gga ggt gct tgg gtg atg      432 
Tyr Val Val Arg Gln Ala Val Leu Ala Lys Gly Gly Ala Trp Val Met 
    130                 135                 140 

tgc att tac gga gtt cct gtg ctg gcc gta aac gca ttc ttt gtt tta      480 
Cys Ile Tyr Gly Val Pro Val Leu Ala Val Asn Ala Phe Phe Val Leu 
145                 150                 155                 160 

atc act tat ctt cac cac acg cat ctc tca ctg ccc cac tat gat agc      528 
Ile Thr Tyr Leu His His Thr His Leu Ser Leu Pro His Tyr Asp Ser 
                165                 170                 175 

tca gaa tgg gac tgg cta cga g                                        550 
Ser Glu Trp Asp Trp Leu Arg 
            180 

 
           
             6  
             183  
             PRT  
             Vernonia galamensis  
           
            6 

His His Ala Phe Ser Asp Tyr Gln Trp Ile Asp Asp Thr Val Gly Phe 
1               5                   10                  15 

Ile Leu His Phe Ala Leu Phe Thr Pro Tyr Phe Ser Trp Lys Tyr Ser 
            20                  25                  30 

His Arg Asn His His Ala Asn Thr Asn Ser Leu Val Thr Asp Glu Val 
        35                  40                  45 

Tyr Ile Pro Lys Val Lys Ser Lys Val Lys Ile Tyr Ser Lys Ile Leu 
    50                  55                  60 

Asn Asn Pro Pro Gly Arg Val Phe Thr Leu Ala Phe Arg Leu Ile Val 
65                  70                  75                  80 

Gly Phe Pro Leu Tyr Leu Phe Thr Asn Val Ser Gly Lys Lys Tyr Glu 
                85                  90                  95 

Arg Phe Ala Asn His Phe Asp Pro Met Ser Pro Ile Phe Thr Glu Arg 
            100                 105                 110 

Glu His Val Gln Val Leu Leu Ser Asp Phe Gly Leu Ile Ala Val Ala 
        115                 120                 125 

Tyr Val Val Arg Gln Ala Val Leu Ala Lys Gly Gly Ala Trp Val Met 
    130                 135                 140 

Cys Ile Tyr Gly Val Pro Val Leu Ala Val Asn Ala Phe Phe Val Leu 
145                 150                 155                 160 

Ile Thr Tyr Leu His His Thr His Leu Ser Leu Pro His Tyr Asp Ser 
                165                 170                 175 

Ser Glu Trp Asp Trp Leu Arg 
                  180 

 
           
             7  
             177  
             DNA  
             Crepis alpina  
             
               CDS  
               (1)..(177)  
                 
             
           
            7 

gaa tgc ggt cac cat gcc ttc agc gac tac cag tgg gtt gac gac aat       48 
Glu Cys Gly His His Ala Phe Ser Asp Tyr Gln Trp Val Asp Asp Asn 
1               5                   10                  15 

gtg ggc ttc atc ctc cac tcg ttt ctc atg acc ccg tat ttc tcc tgg       96 
Val Gly Phe Ile Leu His Ser Phe Leu Met Thr Pro Tyr Phe Ser Trp 
            20                  25                  30 

aaa tac agc cac cgg aac cac cat gcc aac aca aat tcg ctt gac aac      144 
Lys Tyr Ser His Arg Asn His His Ala Asn Thr Asn Ser Leu Asp Asn 
        35                  40                  45 

gat gaa gtt tac atc ccc aaa agc aag gcc aaa                          177 
Asp Glu Val Tyr Ile Pro Lys Ser Lys Ala Lys 
    50                  55 

 
           
             8  
             59  
             PRT  
             Crepis alpina  
           
            8 

Glu Cys Gly His His Ala Phe Ser Asp Tyr Gln Trp Val Asp Asp Asn 
1               5                   10                  15 

Val Gly Phe Ile Leu His Ser Phe Leu Met Thr Pro Tyr Phe Ser Trp 
            20                  25                  30 

Lys Tyr Ser His Arg Asn His His Ala Asn Thr Asn Ser Leu Asp Asn 
        35                  40                  45 

Asp Glu Val Tyr Ile Pro Lys Ser Lys Ala Lys 
    50                  55 

 
           
             9  
             383  
             PRT  
             Arabidopsis thaliana  
           
            9 

Met Gly Ala Gly Gly Arg Met Pro Val Pro Thr Ser Ser Lys Lys Ser 
1               5                   10                  15 

Glu Thr Asp Thr Thr Lys Arg Val Pro Cys Glu Lys Pro Pro Phe Ser 
            20                  25                  30 

Val Gly Asp Leu Lys Lys Ala Ile Pro Pro His Cys Phe Lys Arg Ser 
        35                  40                  45 

Ile Pro Arg Ser Phe Ser Tyr Leu Ile Ser Asp Ile Ile Ile Ala Ser 
    50                  55                  60 

Cys Phe Tyr Tyr Val Ala Thr Asn Tyr Phe Ser Leu Leu Pro Gln Pro 
65                  70                  75                  80 

Leu Ser Tyr Leu Ala Trp Pro Leu Tyr Trp Ala Cys Gln Gly Cys Val 
                85                  90                  95 

Leu Thr Gly Ile Trp Val Ile Ala His Glu Cys Gly His His Ala Phe 
            100                 105                 110 

Ser Asp Tyr Gln Trp Leu Asp Asp Thr Val Gly Leu Ile Phe His Ser 
        115                 120                 125 

Phe Leu Leu Val Pro Tyr Phe Ser Trp Lys Tyr Ser His Arg Arg His 
    130                 135                 140 

His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 
145                 150                 155                 160 

Gln Lys Ser Ala Ile Lys Trp Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 
                165                 170                 175 

Gly Arg Ile Met Met Leu Thr Val Gln Phe Val Leu Gly Trp Pro Leu 
            180                 185                 190 

Tyr Leu Ala Phe Asn Val Ser Gly Arg Pro Tyr Asp Gly Phe Ala Cys 
        195                 200                 205 

His Phe Phe Pro Asn Ala Pro Ile Tyr Asn Asp Arg Glu Arg Leu Gln 
    210                 215                 220 

Ile Tyr Leu Ser Asp Ala Gly Ile Leu Ala Val Cys Phe Gly Leu Tyr 
225                 230                 235                 240 

Arg Tyr Ala Ala Ala Gln Gly Met Ala Ser Met Ile Cys Leu Tyr Gly 
                245                 250                 255 

Val Pro Leu Leu Ile Val Asn Ala Phe Leu Val Leu Ile Thr Tyr Leu 
            260                 265                 270 

Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Trp Asp 
        275                 280                 285 

Trp Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly Ile Leu 
    290                 295                 300 

Asn Lys Val Phe His Asn Ile Thr Asp Thr His Val Ala His His Leu 
305                 310                 315                 320 

Phe Ser Thr Met Pro His Tyr Asn Ala Met Glu Ala Thr Lys Ala Ile 
                325                 330                 335 

Lys Pro Ile Leu Gly Asp Tyr Tyr Gln Phe Asp Gly Thr Pro Trp Tyr 
            340                 345                 350 

Val Ala Met Tyr Arg Glu Ala Lys Glu Cys Ile Tyr Val Glu Pro Asp 
        355                 360                 365 

Arg Glu Gly Asp Lys Lys Gly Val Tyr Trp Tyr Asn Asn Lys Leu 
    370                 375                 380 

 
           
             10  
             384  
             PRT  
             Brassica juncea  
           
            10 

Met Gly Ala Gly Gly Arg Met Gln Val Ser Pro Ser Pro Lys Lys Ser 
1               5                   10                  15 

Glu Thr Asp Thr Leu Lys Arg Val Pro Cys Glu Thr Pro Pro Phe Thr 
            20                  25                  30 

Val Gly Glu Leu Lys Lys Ala Ile Pro Pro His Cys Phe Lys Arg Ser 
        35                  40                  45 

Ile Pro Arg Ser Phe Ser Tyr Leu Ile Trp Asp Ile Ile Val Ala Ser 
    50                  55                  60 

Cys Phe Tyr Tyr Val Ala Thr Thr Tyr Phe Pro Leu Leu Pro His Pro 
65                  70                  75                  80 

Leu Ser Tyr Val Ala Trp Pro Leu Tyr Trp Ala Cys Gln Gly Val Val 
                85                  90                  95 

Leu Thr Gly Val Trp Val Ile Ala His Glu Cys Gly His His Ala Phe 
            100                 105                 110 

Ser Asp Tyr Gln Trp Leu Asp Asp Thr Val Gly Leu Ile Phe His Ser 
        115                 120                 125 

Phe Leu Leu Val Pro Tyr Phe Ser Trp Lys Tyr Ser His Arg Arg His 
    130                 135                 140 

His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 
145                 150                 155                 160 

Lys Lys Ser Asp Ile Lys Trp Tyr Gly Lys Tyr Leu Asn Asn Pro Leu 
                165                 170                 175 

Gly Arg Thr Val Met Leu Thr Val Gln Phe Thr Leu Gly Trp Pro Leu 
            180                 185                 190 

Tyr Trp Ala Phe Asn Val Ser Gly Arg Pro Tyr Pro Glu Gly Phe Ala 
        195                 200                 205 

Cys His Phe His Pro Asn Ala Pro Ile Tyr Asn Asp Arg Glu Arg Leu 
    210                 215                 220 

Gln Ile Tyr Val Ser Asp Ala Gly Ile Leu Ala Val Cys Tyr Gly Leu 
225                 230                 235                 240 

Tyr Arg Tyr Ala Ala Ala Gln Gly Val Ala Ser Met Val Cys Leu Tyr 
                245                 250                 255 

Gly Val Pro Leu Leu Ile Val Asn Ala Phe Leu Val Leu Ile Thr Tyr 
            260                 265                 270 

Leu Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Ser Glu Trp 
        275                 280                 285 

Asp Trp Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly Ile 
    290                 295                 300 

Leu Asn Lys Val Phe His Asn Ile Thr Asp Thr His Val Ala His His 
305                 310                 315                 320 

Leu Phe Ser Thr Met Pro His Tyr His Ala Met Glu Val Thr Lys Ala 
                325                 330                 335 

Ile Lys Pro Ile Leu Gly Asp Tyr Tyr Gln Phe Asp Gly Thr Pro Trp 
            340                 345                 350 

Val Lys Ala Met Trp Arg Glu Ala Lys Glu Cys Ile Tyr Val Glu Pro 
        355                 360                 365 

Asp Arg Gln Gly Glu Lys Lys Gly Val Phe Trp Tyr Asn Asn Lys Leu 
    370                 375                 380 

 
           
             11  
             367  
             PRT  
             Glycine max  
           
            11 

Met Gly Ala Gly Gly Arg Thr Asp Val Pro Pro Ala Asn Arg Lys Ser 
1               5                   10                  15 

Glu Val Asp Pro Leu Lys Arg Val Pro Phe Glu Lys Pro Gln Phe Ser 
            20                  25                  30 

Leu Ser Gln Ile Lys Lys Ala Ile Pro Pro His Cys Phe Gln Arg Ser 
        35                  40                  45 

Val Leu Arg Ser Phe Ser Tyr Val Val Tyr Asp Leu Thr Ile Ala Phe 
    50                  55                  60 

Cys Leu Tyr Tyr Val Ala Thr His Tyr Phe His Leu Leu Pro Gly Pro 
65                  70                  75                  80 

Leu Ser Phe Arg Gly Met Ala Ile Tyr Trp Ala Val Gln Gly Cys Ile 
                85                  90                  95 

Leu Thr Gly Val Trp Val Ile Ala His Glu Cys Gly His His Ala Phe 
            100                 105                 110 

Ser Asp Tyr Gln Leu Leu Asp Asp Ile Val Gly Leu Ile Leu His Ser 
        115                 120                 125 

Ala Leu Leu Val Pro Tyr Phe Ser Trp Lys Tyr Ser His Arg Arg His 
    130                 135                 140 

His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 
145                 150                 155                 160 

Gly Arg Val Leu Thr Leu Ala Val Thr Leu Thr Leu Gly Trp Pro Leu 
                165                 170                 175 

Tyr Leu Ala Leu Asn Val Ser Gly Arg Pro Tyr Asp Arg Phe Ala Cys 
            180                 185                 190 

His Tyr Asp Pro Tyr Gly Pro Ile Tyr Ser Asp Arg Glu Arg Leu Gln 
        195                 200                 205 

Ile Tyr Ile Ser Asp Ala Gly Val Leu Ala Val Val Tyr Gly Leu Phe 
    210                 215                 220 

Arg Leu Ala Met Ala Lys Gly Leu Ala Trp Val Val Cys Val Tyr Gly 
225                 230                 235                 240 

Val Pro Leu Leu Val Val Asn Gly Phe Leu Val Leu Ile Thr Phe Leu 
                245                 250                 255 

Gln His Thr His Pro Ala Leu Pro His Tyr Thr Ser Ser Glu Trp Asp 
            260                 265                 270 

Trp Leu Arg Gly Ala Leu Ala Thr Val Asp Arg Asp Tyr Gly Ile Leu 
        275                 280                 285 

Asn Lys Val Phe His Asn Ile Thr Asp Thr His Val Ala His His Leu 
    290                 295                 300 

Phe Ser Thr Met Pro His Tyr His Ala Met Glu Ala Thr Lys Ala Ile 
305                 310                 315                 320 

Lys Pro Ile Leu Gly Glu Tyr Tyr Arg Phe Asp Glu Thr Pro Phe Val 
                325                 330                 335 

Lys Ala Met Trp Arg Glu Ala Arg Glu Cys Ile Tyr Val Glu Pro Asp 
            340                 345                 350 

Gln Ser Thr Glu Ser Lys Gly Val Phe Trp Tyr Asn Asn Lys Leu 
        355                 360                 365 

 
           
             12  
             383  
             PRT  
             Solanum commersonii  
           
            12 

Met Gly Ala Gly Gly Arg Met Ser Ala Pro Asn Gly Glu Thr Glu Val 
1               5                   10                  15 

Lys Arg Asn Pro Leu Gln Lys Val Pro Thr Ser Lys Pro Pro Phe Thr 
            20                  25                  30 

Val Gly Asp Ile Lys Lys Ala Ile Pro Pro His Cys Phe Gln Arg Ser 
        35                  40                  45 

Leu Ile Arg Ser Phe Ser Tyr Val Val Tyr Asp Leu Ile Leu Val Ser 
    50                  55                  60 

Ile Met Tyr Tyr Val Ala Asn Thr Tyr Phe His Leu Leu Pro Ser Pro 
65                  70                  75                  80 

Tyr Cys Tyr Ile Ala Trp Pro Ile Tyr Trp Ile Cys Gln Gly Cys Val 
                85                  90                  95 

Cys Thr Gly Ile Trp Val Asn Ala His Glu Cys Gly His His Ala Phe 
            100                 105                 110 

Ser Asp Tyr Gln Trp Val Asp Asp Thr Val Gly Leu Ile Leu His Ser 
        115                 120                 125 

Ala Leu Leu Val Pro Tyr Phe Ser Trp Lys Tyr Ser His Arg Arg His 
    130                 135                 140 

His Ser Asn Thr Gly Ser Leu Glu Arg Asp Glu Val Phe Val Pro Lys 
145                 150                 155                 160 

Pro Lys Ser Gln Leu Gly Trp Tyr Ser Lys Tyr Leu Asn Asn Pro Pro 
                165                 170                 175 

Gly Arg Val Leu Ser Leu Thr Ile Thr Leu Thr Leu Gly Trp Pro Leu 
            180                 185                 190 

Tyr Leu Ala Phe Asn Val Ser Gly Arg Pro Tyr Asp Arg Phe Ala Cys 
        195                 200                 205 

His Tyr Asp Pro Tyr Gly Pro Ile Tyr Asn Asn Arg Glu Arg Leu Gln 
    210                 215                 220 

Ile Phe Ile Ser Asp Ala Gly Val Leu Gly Val Cys Tyr Leu Leu Tyr 
225                 230                 235                 240 

Arg Ile Ala Leu Val Lys Gly Leu Ala Trp Leu Val Cys Val Tyr Gly 
                245                 250                 255 

Val Pro Leu Leu Val Val Asn Gly Phe Leu Val Leu Ile Thr Tyr Leu 
            260                 265                 270 

Gln His Thr His Pro Ser Leu Pro His Tyr Asp Ser Thr Glu Trp Asp 
        275                 280                 285 

Trp Leu Arg Gly Ala Leu Ala Thr Cys Asp Arg Asp Tyr Gly Val Leu 
    290                 295                 300 

Asn Lys Val Phe His Asn Ile Thr Asp Thr His Val Val His His Leu 
305                 310                 315                 320 

Phe Ser Thr Met Pro His Tyr Asn Ala Met Glu Ala Thr Lys Ala Val 
                325                 330                 335 

Lys Pro Leu Leu Gly Asp Tyr Tyr Gln Phe Asp Gly Thr Pro Ile Tyr 
            340                 345                 350 

Lys Glu Met Trp Arg Glu Ala Lys Glu Cys Leu Tyr Val Glu Lys Asp 
        355                 360                 365 

Glu Ser Ser Gln Gly Lys Gly Val Phe Trp Tyr Lys Asn Lys Leu 
    370                 375                 380 

 
           
             13  
             387  
             PRT  
             Glycine max  
           
            13 

Met Gly Leu Ala Lys Glu Thr Thr Met Gly Gly Arg Gly Arg Val Ala 
1               5                   10                  15 

Lys Val Glu Val Gln Gly Lys Lys Pro Leu Ser Arg Val Pro Asn Thr 
            20                  25                  30 

Lys Pro Pro Phe Thr Val Gly Gln Leu Lys Lys Ala Ile Pro Pro His 
        35                  40                  45 

Cys Phe Gln Arg Ser Leu Leu Thr Ser Phe Ser Tyr Val Val Tyr Asp 
    50                  55                  60 

Leu Ser Phe Ala Phe Ile Phe Tyr Ile Ala Thr Thr Tyr Phe His Leu 
65                  70                  75                  80 

Leu Pro Gln Pro Phe Ser Leu Ile Ala Trp Pro Ile Tyr Trp Val Leu 
                85                  90                  95 

Gln Gly Cys Leu Leu Thr Gly Val Trp Val Ile Ala His Glu Cys Gly 
            100                 105                 110 

His His Ala Phe Ser Lys Tyr Gln Trp Val Asp Asp Val Val Gly Leu 
        115                 120                 125 

Thr Leu His Ser Thr Leu Leu Val Pro Tyr Phe Ser Trp Lys Ile Ser 
    130                 135                 140 

His Arg Arg His His Ser Asn Thr Gly Ser Leu Asp Arg Asp Glu Val 
145                 150                 155                 160 

Phe Val Pro Lys Pro Lys Ser Lys Val Ala Trp Phe Ser Lys Tyr Leu 
                165                 170                 175 

Asn Asn Pro Leu Gly Arg Ala Val Ser Leu Leu Val Thr Leu Thr Ile 
            180                 185                 190 

Gly Trp Pro Met Tyr Leu Ala Phe Asn Val Ser Gly Arg Pro Tyr Asp 
        195                 200                 205 

Ser Phe Ala Ser His Tyr His Pro Tyr Ala Pro Ile Tyr Ser Asn Arg 
    210                 215                 220 

Glu Arg Leu Leu Ile Tyr Val Ser Asp Val Ala Leu Phe Ser Val Thr 
225                 230                 235                 240 

Tyr Ser Leu Tyr Arg Val Ala Thr Leu Lys Gly Leu Val Trp Leu Leu 
                245                 250                 255 

Cys Val Tyr Gly Val Pro Leu Leu Ile Val Asn Gly Phe Leu Val Thr 
            260                 265                 270 

Ile Thr Tyr Leu Gln His Thr His Phe Ala Leu Pro His Tyr Asp Ser 
        275                 280                 285 

Ser Glu Trp Asp Trp Leu Lys Gly Ala Leu Ala Thr Met Asp Arg Asp 
    290                 295                 300 

Tyr Gly Ile Leu Asn Lys Val Phe His His Ile Thr Asp Thr His Val 
305                 310                 315                 320 

Ala His His Leu Phe Ser Thr Met Pro His Tyr His Ala Met Glu Ala 
                325                 330                 335 

Thr Asn Ala Ile Lys Pro Ile Leu Gly Glu Tyr Tyr Gln Phe Asp Asp 
            340                 345                 350 

Thr Pro Phe Tyr Lys Ala Leu Trp Arg Glu Ala Arg Glu Cys Leu Tyr 
        355                 360                 365 

Val Glu Pro Asp Glu Gly Thr Ser Glu Lys Gly Val Tyr Trp Tyr Arg 
    370                 375                 380 

Asn Lys Tyr 
385 

 
           
             14  
             387  
             PRT  
             Ricinus communis  
           
            14 

Met Gly Gly Gly Gly Arg Met Ser Thr Val Ile Thr Ser Asn Asn Ser 
1               5                   10                  15 

Glu Lys Lys Gly Gly Ser Ser His Leu Lys Arg Ala Pro His Thr Lys 
            20                  25                  30 

Pro Pro Phe Thr Leu Gly Asp Leu Lys Arg Ala Ile Pro Pro His Cys 
        35                  40                  45 

Phe Glu Arg Ser Phe Val Arg Ser Phe Ser Tyr Val Ala Tyr Asp Val 
    50                  55                  60 

Cys Leu Ser Phe Leu Phe Tyr Ser Ile Ala Thr Asn Phe Phe Pro Tyr 
65                  70                  75                  80 

Ile Ser Ser Pro Leu Ser Tyr Val Ala Trp Leu Val Tyr Trp Leu Phe 
                85                  90                  95 

Gln Gly Cys Ile Leu Thr Gly Leu Trp Val Ile Gly His Glu Cys Gly 
            100                 105                 110 

His His Ala Phe Ser Glu Tyr Gln Leu Ala Asp Asp Ile Val Gly Leu 
        115                 120                 125 

Ile Val His Ser Ala Leu Leu Val Pro Tyr Phe Ser Trp Lys Tyr Ser 
    130                 135                 140 

His Arg Arg His His Ser Asn Ile Gly Ser Leu Glu Arg Asp Glu Val 
145                 150                 155                 160 

Phe Val Pro Lys Ser Lys Ser Lys Ile Ser Trp Tyr Ser Lys Tyr Ser 
                165                 170                 175 

Asn Asn Pro Pro Gly Arg Val Leu Thr Leu Ala Ala Thr Leu Leu Leu 
            180                 185                 190 

Gly Trp Pro Leu Tyr Leu Ala Phe Asn Val Ser Gly Arg Pro Tyr Asp 
        195                 200                 205 

Arg Phe Ala Cys His Tyr Asp Pro Tyr Gly Pro Ile Phe Ser Glu Arg 
    210                 215                 220 

Glu Arg Leu Gln Ile Tyr Ile Ala Asp Leu Gly Ile Phe Ala Thr Thr 
225                 230                 235                 240 

Phe Val Leu Tyr Gln Ala Thr Met Ala Lys Gly Leu Ala Trp Val Met 
                245                 250                 255 

Arg Ile Tyr Gly Val Pro Leu Leu Ile Val Asn Cys Phe Leu Val Met 
            260                 265                 270 

Ile Thr Tyr Leu Gln His Thr His Pro Ala Ile Pro Arg Tyr Gly Ser 
        275                 280                 285 

Ser Glu Trp Asp Trp Leu Arg Gly Ala Met Val Thr Val Asp Arg Asp 
    290                 295                 300 

Tyr Gly Val Leu Asn Lys Val Phe His Asn Ile Ala Asp Thr His Val 
305                 310                 315                 320 

Ala His His Leu Phe Ala Thr Val Pro His Tyr His Ala Met Glu Ala 
                325                 330                 335 

Thr Lys Ala Ile Lys Pro Ile Met Gly Glu Tyr Tyr Arg Tyr Asp Gly 
            340                 345                 350 

Thr Pro Phe Tyr Lys Ala Leu Trp Arg Glu Ala Lys Glu Cys Leu Phe 
        355                 360                 365 

Val Glu Pro Asp Glu Gly Ala Pro Thr Gln Gly Val Phe Trp Tyr Arg 
    370                 375                 380 

Asn Lys Tyr 
385 

 
           
             15  
             6  
             PRT  
             mixed function monooxygenase peptide motif  
           
            15 

His Glu Cys Gly His His 
1               5 

 
           
             16  
             5  
             PRT  
             mixed function monooxygenase peptide motif  
           
            16 

His Arg Asn His His 
1               5 

 
           
             17  
             5  
             PRT  
             mixed function monooxygenase peptide motif  
           
            17 

His Val Met His His 
1               5 

 
           
             18  
             5  
             PRT  
             mixed function monooxygenase peptide motif  
           
            18 

His Val Leu His His 
1               5 

 
           
             19  
             1199  
             DNA  
             Vernonia galamensis  
             
               CDS  
               (44)..(1195)  
                 
             
           
            19 

tattacacat ttacactgat ctgttaatca aatttcaaac aaa atg gga gct ggt       55 
                                                Met Gly Ala Gly 
                                                1 

ggc cga atg aat acc acc gat gat gat cag aag aat ctc ttc caa cgc      103 
Gly Arg Met Asn Thr Thr Asp Asp Asp Gln Lys Asn Leu Phe Gln Arg 
5                   10                  15                  20 

gta cca gcc tcc aaa cca cca ttc tcc ttg gct gat ctt aag aaa gcc      151 
Val Pro Ala Ser Lys Pro Pro Phe Ser Leu Ala Asp Leu Lys Lys Ala 
                25                  30                  35 

ata cca ccc cac tgt ttc caa aga tcc ctc ctc cgt tca tct tac tat      199 
Ile Pro Pro His Cys Phe Gln Arg Ser Leu Leu Arg Ser Ser Tyr Tyr 
            40                  45                  50 

gtg gtt cat gat ctc gtc gta gcc tac gtc ttt tac tat ctc gcc aac      247 
Val Val His Asp Leu Val Val Ala Tyr Val Phe Tyr Tyr Leu Ala Asn 
        55                  60                  65 

aca tac atc cct ctt ctt ccc tcc cct ctt gcc tac tta tta gct tgg      295 
Thr Tyr Ile Pro Leu Leu Pro Ser Pro Leu Ala Tyr Leu Leu Ala Trp 
    70                  75                  80 

ccc ctt tac tgg ttc tgt cag ggt agc atc ctc acc ggt gtc tgg gtc      343 
Pro Leu Tyr Trp Phe Cys Gln Gly Ser Ile Leu Thr Gly Val Trp Val 
85                  90                  95                  100 

atc ggt cat gaa tgt ggc cac cat gcc ttc agt gac tat caa tgg ata      391 
Ile Gly His Glu Cys Gly His His Ala Phe Ser Asp Tyr Gln Trp Ile 
                105                 110                 115 

gac gac act gtg ggc ttc atc ctt cac tct gca ctc ttc acc cct tat      439 
Asp Asp Thr Val Gly Phe Ile Leu His Ser Ala Leu Phe Thr Pro Tyr 
            120                 125                 130 

ttc tct tgg aaa tac agt cac cgt aat cac cat gcc aac aca aac tct      487 
Phe Ser Trp Lys Tyr Ser His Arg Asn His His Ala Asn Thr Asn Ser 
        135                 140                 145 

ctt gat aac gat gaa gta tac atc cct aaa gtt aaa tcc aag gtc aag      535 
Leu Asp Asn Asp Glu Val Tyr Ile Pro Lys Val Lys Ser Lys Val Lys 
    150                 155                 160 

att tat tcc aaa atc ctt aac aac cct cct ggt cgc gtt ttc acc ttg      583 
Ile Tyr Ser Lys Ile Leu Asn Asn Pro Pro Gly Arg Val Phe Thr Leu 
165                 170                 175                 180 

gct ttc aga ttg atc gtg ggt ttt cct tta tac ctt ttc acc aat gtt      631 
Ala Phe Arg Leu Ile Val Gly Phe Pro Leu Tyr Leu Phe Thr Asn Val 
                185                 190                 195 

tca ggc aag aaa tac gaa cgt ttt gcc aac cat ttt gat ccc atg agt      679 
Ser Gly Lys Lys Tyr Glu Arg Phe Ala Asn His Phe Asp Pro Met Ser 
            200                 205                 210 

ccc att ttc acc gag cgt gag cat gta caa gtc ttg ctt tct gat ttt      727 
Pro Ile Phe Thr Glu Arg Glu His Val Gln Val Leu Leu Ser Asp Phe 
        215                 220                 225 

ggt ctc ata gca gtt gct tac gtg gtt cgt caa gct gta ctg gct aaa      775 
Gly Leu Ile Ala Val Ala Tyr Val Val Arg Gln Ala Val Leu Ala Lys 
    230                 235                 240 

gga ggt gct tgg gtg atg tgc att tac gga gtt cct gtg ctg gcc gta      823 
Gly Gly Ala Trp Val Met Cys Ile Tyr Gly Val Pro Val Leu Ala Val 
245                 250                 255                 260 

aac gca ttc ttt gtt tta atc act tat ctt cac cac acg cat ctc tca      871 
Asn Ala Phe Phe Val Leu Ile Thr Tyr Leu His His Thr His Leu Ser 
                265                 270                 275 

ctg cct cac tat gat tcg act gaa tgg gac tgg atc aag gga gct ttg      919 
Leu Pro His Tyr Asp Ser Thr Glu Trp Asp Trp Ile Lys Gly Ala Leu 
            280                 285                 290 

tgc acc atc gac aga gat ttc gga ttc ttg aat agg gtt ttc cac gac      967 
Cys Thr Ile Asp Arg Asp Phe Gly Phe Leu Asn Arg Val Phe His Asp 
        295                 300                 305 

gtg aca cac acc cat gtg ttg cat cat ttg ata tcg tac att cct cat     1015 
Val Thr His Thr His Val Leu His His Leu Ile Ser Tyr Ile Pro His 
    310                 315                 320 

tat cat gca aag gag gca aga gac gcc atc aaa ccg gtg ttg ggc gaa     1063 
Tyr His Ala Lys Glu Ala Arg Asp Ala Ile Lys Pro Val Leu Gly Glu 
325                 330                 335                 340 

tac tat aag atc gac agg aca ccg atc gtg aag gca atg tgg agg gaa     1111 
Tyr Tyr Lys Ile Asp Arg Thr Pro Ile Val Lys Ala Met Trp Arg Glu 
                345                 350                 355 

gca aag aat gca tat aca ttg agg ctg atg aag ata gcg agc acc aag     1159 
Ala Lys Asn Ala Tyr Thr Leu Arg Leu Met Lys Ile Ala Ser Thr Lys 
            360                 365                 370 

gca cat act ggt acc aca agt tgt aaa gcc aga tcc taag                1199 
Ala His Thr Gly Thr Thr Ser Cys Lys Ala Arg Ser 
        375                 380 

 
           
             20  
             384  
             PRT  
             Vernonia galamensis  
           
            20 

Met Gly Ala Gly Gly Arg Met Asn Thr Thr Asp Asp Asp Gln Lys Asn 
1               5                   10                  15 

Leu Phe Gln Arg Val Pro Ala Ser Lys Pro Pro Phe Ser Leu Ala Asp 
            20                  25                  30 

Leu Lys Lys Ala Ile Pro Pro His Cys Phe Gln Arg Ser Leu Leu Arg 
        35                  40                  45 

Ser Ser Tyr Tyr Val Val His Asp Leu Val Val Ala Tyr Val Phe Tyr 
    50                  55                  60 

Tyr Leu Ala Asn Thr Tyr Ile Pro Leu Leu Pro Ser Pro Leu Ala Tyr 
65                  70                  75                  80 

Leu Leu Ala Trp Pro Leu Tyr Trp Phe Cys Gln Gly Ser Ile Leu Thr 
                85                  90                  95 

Gly Val Trp Val Ile Gly His Glu Cys Gly His His Ala Phe Ser Asp 
            100                 105                 110 

Tyr Gln Trp Ile Asp Asp Thr Val Gly Phe Ile Leu His Ser Ala Leu 
        115                 120                 125 

Phe Thr Pro Tyr Phe Ser Trp Lys Tyr Ser His Arg Asn His His Ala 
    130                 135                 140 

Asn Thr Asn Ser Leu Asp Asn Asp Glu Val Tyr Ile Pro Lys Val Lys 
145                 150                 155                 160 

Ser Lys Val Lys Ile Tyr Ser Lys Ile Leu Asn Asn Pro Pro Gly Arg 
                165                 170                 175 

Val Phe Thr Leu Ala Phe Arg Leu Ile Val Gly Phe Pro Leu Tyr Leu 
            180                 185                 190 

Phe Thr Asn Val Ser Gly Lys Lys Tyr Glu Arg Phe Ala Asn His Phe 
        195                 200                 205 

Asp Pro Met Ser Pro Ile Phe Thr Glu Arg Glu His Val Gln Val Leu 
    210                 215                 220 

Leu Ser Asp Phe Gly Leu Ile Ala Val Ala Tyr Val Val Arg Gln Ala 
225                 230                 235                 240 

Val Leu Ala Lys Gly Gly Ala Trp Val Met Cys Ile Tyr Gly Val Pro 
                245                 250                 255 

Val Leu Ala Val Asn Ala Phe Phe Val Leu Ile Thr Tyr Leu His His 
            260                 265                 270 

Thr His Leu Ser Leu Pro His Tyr Asp Ser Thr Glu Trp Asp Trp Ile 
        275                 280                 285 

Lys Gly Ala Leu Cys Thr Ile Asp Arg Asp Phe Gly Phe Leu Asn Arg 
    290                 295                 300 

Val Phe His Asp Val Thr His Thr His Val Leu His His Leu Ile Ser 
305                 310                 315                 320 

Tyr Ile Pro His Tyr His Ala Lys Glu Ala Arg Asp Ala Ile Lys Pro 
                325                 330                 335 

Val Leu Gly Glu Tyr Tyr Lys Ile Asp Arg Thr Pro Ile Val Lys Ala 
            340                 345                 350 

Met Trp Arg Glu Ala Lys Asn Ala Tyr Thr Leu Arg Leu Met Lys Ile 
        355                 360                 365 

Ala Ser Thr Lys Ala His Thr Gly Thr Thr Ser Cys Lys Ala Arg Ser 
    370                 375                 380 

 
           
             21  
             5  
             PRT  
             mixed function monooxygenase consensus motif  
             
               MISC_FEATURE  
               (2)..(4)  
               Xaa at position 2 is any amino acid; Xaa at 
      position 3 is any amino acid; Xaa at position 4 is any amino acid;  
             
           
            21 

His Xaa Xaa Xaa His 
1               5 

 
           
             22  
             6  
             PRT  
             mixed function monooxygenase consensus motif  
             
               MISC_FEATURE  
               (2)..(5)  
               Xaa at position 2 is any amino acid; Xaa at 
      position 3 is any amino acid; Xaa at position 4 is any amino 
      acid; Xaa at position 5 is any amino acid;  
             
           
            22 

His Xaa Xaa Xaa Xaa His 
1               5 

 
           
             23  
             5  
             PRT  
             mixed function monooxygenase consensus motif  
             
               MISC_FEATURE  
               (2)..(3)  
               Xaa at position 2 is any amino acid; Xaa at 
      position 3 is any amino acid;  
             
           
            23 

His Xaa Xaa His His 
1               5 

 
           
             24  
             6  
             PRT  
             mixed function monooxygenase consensus motif  
             
               MISC_FEATURE  
               (2)..(4)  
               Xaa at position 2 is any amino acid; Xaa at 
      position 3 is any amino acid; Xaa at position 4 is any amino acid;  
             
           
            24 

His Xaa Xaa Xaa His His 
1               5