US2003213010A1PendingUtilityA1

Transgenic high tryptophan plants

52
Priority: May 3, 2002Filed: May 5, 2003Published: Nov 13, 2003
Est. expiryMay 3, 2022(expired)· nominal 20-yr term from priority
Inventors:Lisa Marie WeaverTim N. OulmassovGabriela VaduvaJihong LiangMarguerite VaragonaTyamagondlu V. Venkatesh
C12N 9/88C12N 15/8254
52
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Claims

Abstract

The present invention provides a method for altering the tryptophan content of a plant by introducing and expressing an isolated DNA segment encoding an anthranilate synthase in the cells of the plant. Transgenic plants transformed with an isolated DNA segment encoding an anthranilate synthase, as well as human or animal food, seeds and progeny derived from these plants, are also provided.

Claims

exact text as granted — not AI-modified
What is claimed:  
     
         1 . An isolated DNA molecule encoding an anthranilate synthase, wherein the DNA molecule encodes a protein substantially homologous to a protein selected from the group consisting of SEQ ID NOs: 66, 108, 109, 110, 111, 133, and 137.  
     
     
         2 . The isolated DNA molecule of  claim 1 , wherein the DNA molecule is substantially homologous to a DNA molecule selected from a group consisting of SEQ ID NOs: 67, 68, 104, 105, 106, 107, 134, 135, and 136.  
     
     
         3 . A DNA construct comprising an expression cassette, wherein the expression cassette in operable linkage comprises (i) a heterologous promoter; (ii) a DNA molecule encoding a monomeric anthranilate synthase protein, wherein the monomeric anthranilate synthase comprises a single polypeptide comprising an anthranilate synthase α-domain and an anthranilate synthase β-domain; and (iii) a transcriptional terminator.  
     
     
         4 . The DNA construct of  claim 3 , wherein the monomeric anthranilate synthase protein is substantially homologous to a protein selected from the group consisting of SEQ ID NOs: 4, 7, 43, 57, 77, 78, 79, 80, 81, 82, 130, 131, and 132.  
     
     
         5 . The DNA construct of  claim 3 , wherein the protein is SEQ ID NO: 4.  
     
     
         6 . The DNA construct of  claim 3 , wherein the DNA molecule is substantially homologous to a DNA molecule selected from the group consisting of SEQ ID NOs: 1, 75, 76, 83, 121, 122, 123, 124, 125, 126, 127, 128, and 129.  
     
     
         7 . The DNA construct of  claim 3 , wherein the expression cassette further encodes a plastid transit peptide.  
     
     
         8 . The DNA construct of  claim 7 , wherein the plastid transit peptide is selected from the group consisting of SEQ ID NOs: 72, 74, 114, and 115.  
     
     
         9 . A DNA construct comprising a first expression cassette, wherein the first expression cassette in operable linkage comprises (i) a heterologous promoter; (ii) a DNA molecule encoding an anthranilate synthase α-domain protein; and (iii) a transcriptional terminator.  
     
     
         10 . The DNA construct of  claim 9 , further comprising a second expression cassette in operable linkage comprising (i) a heterologous promoter; (ii) a DNA molecule encoding an anthranilate synthase β-domain protein; and (iii) a transcriptional terminator.  
     
     
         11 . The DNA construct of  claim 10 , wherein the α-domain or β-domain protein is substantially homologous to a protein selected from the group consisting of SEQ ID NOs: 5, 6, 8, 44, 45, 66, 99, 100, 101, 102, 103, 108, 109, 110, 111, 117, 118, 133, and 137.  
     
     
         12 . The DNA construct of  claim 10 , wherein the DNA molecule encoding an anthranilate synthase α-domain or β-domain protein is substantially homologous to a DNA molecule selected from the group consisting of SEQ ID NOs: 2, 3, 67, 94, 95, 96, 97, 98, 104, 105, 106, 112, 116, 119, 120, 134, 135, and 136.  
     
     
         13 . The DNA construct of  claim 10 , wherein the DNA molecule encoding an anthranilate synthase α-domain or β-domain protein is selected from the group consisting of SEQ ID NOs: 2, 3, 67, 94, 95, 96, 97, 98, 104, 105, 106, 112, 116, 119, 120, 134, 135, and 136.  
     
     
         14 . The DNA construct of  claim 10 , wherein the α-domain protein is SEQ ID NO: 66 and the β-domain protein is SEQ ID NO: 118.  
     
     
         15 . A method for altering the tryptophan content in a plant comprising: 
 a. introducing into regenerable cells of a plant a transgene comprising an isolated DNA molecule encoding an anthranilate synthase wherein the isolated DNA is operably linked to a promoter functional in a plant cell, to yield transformed plant cells; and    b. regenerating a plant from the transformed plant cells wherein the cells of the plant express the anthranilate synthase encoded by the isolated DNA in an amount effective to increase the tryptophan content in the plant relative to the tryptophan content in a second plant of the same or similar genetic background that does not include the transgene.    
     
     
         16 . The method of  claim 15 , wherein the anthranilate synthase is substantially homologous to a monomeric anthranilate synthase selected from the group consisting of  Agrobacterium tumefaciens , Anabaena M22983 , Azospirillum brasilense, Bradyrhizobium japonicum, Brucella melitensis, Mesorhizobium loti,  Nostoc sp. PCC7120 , Rhizobium meliloti, Rhodospirillum rubrum, Rhodopseudomonas palustris,  and  Thermobifida fusca.    
     
     
         17 . The method of  claim 15 , wherein the anthranilate synthase is selected from the group consisting of SEQ ID NOs: 4, 7, 43, 57, 77, 78, 79, 80, 81, 82, 130, 131, and 132.  
     
     
         18 . The method of  claim 15 , wherein the anthranilate synthase is encoded by a DNA molecule substantially homologous to a DNA molecule selected from the group consisting of SEQ ID NOs: 1, 75, 76, 83, 121, 122, 123, 124, 125, 126, 127, 128, and 129.  
     
     
         19 . The method of  claim 15 , wherein the anthranilate synthase comprises a chimeric monomeric anthranilate synthase comprising a fusion of an anthranilate synthase α-domain of a first species and an anthranilate synthase β-domain from a second species, wherein the first species and the second species are selected from the group consisting of  Agrobacterium tumefaciens,  Anabaena M22983,  Arabidopsis thaliana, Azospirillum brasilense, Bradyrhizobium japonicum, Brucella melitensis, Escherichia coli, Euglena gracilis, Mesorhizobium loti,  Nostoc sp. PCC7120 , Rhizobium meliloti, Rhodospirillum rubrum, Ruta graveolens, Rhodopseudomonas palustris, Salmonella typhimurium, Serratia marcescens, Sulfolobus solfataricus, Thermobifida fusca,  sorghum, soybean, rice, cotton, wheat, tobacco, and  Zea mays.    
     
     
         20 . The method of  claim 19 , wherein the α-domain or the β-domain is at least a portion of SEQ ID NOs: 4, 5, 6, 7, 8, 43, 44, 45, 57, 66, 77, 78, 79, 80, 81, 82, 99, 100, 101, 102, 103, 108, 109, 110, 111, 117, 118, 130, 131, 132, 133, and 137.  
     
     
         21 . The method of  claim 15 , wherein the DNA molecule further encodes a plastid transit peptide.  
     
     
         22 . The method of  claim 21 , wherein the plastid transit peptide is selected from the group consisting of SEQ ID NOs: 72, 74, 114, and 115.  
     
     
         23 . The method of  claim 15 , wherein the plant is a dicot.  
     
     
         24 . The method of  claim 33 , wherein the dicot is selected from the group consisting of soybean, cotton, and canola.  
     
     
         25 . The method of  claim 20 , wherein the plant is a monocot.  
     
     
         26 . The method of  claim 25 , wherein the monocot is selected from the group consisting of maize, rice, wheat, barley, and sorghum.  
     
     
         27 . A method for making an animal feed or a human food comprising: 
 a. introducing into regenerable cells of a plant a transgene comprising an isolated DNA molecule encoding an anthranilate synthase wherein the isolated DNA molecule is operably linked to a promoter functional in a plant cell, to yield transformed plant cells; and    b. regenerating a plant from the transformed plant cells wherein the cells of the plant express the anthranilate synthase encoded by the isolated DNA molecule in an amount effective to increase the tryptophan content in the plant relative to the tryptophan content in a second plant of the same or similar genetic background that does not include the transgene.    
     
     
         28 . The method of  claim 27 , wherein the anthranilate synthase is selected from the group consisting of SEQ ID NOs: 4, 7, 43, 57, 77, 78, 79, 80, 81, 82, 130, 131, and 132.  
     
     
         29 . The method of  claim 27 , wherein the anthranilate synthase comprises a chimeric monomeric anthranilate synthase comprising a fusion of an anthranilate synthase α-domain of a first species and an anthranilate synthase β-domain from a second species, wherein the first species and the second species are selected from the group consisting of  Agrobacterium tumefaciens,  Anabaena M22983,  Arabidopsis thaliana, Azospirillum brasilense, Bradyrhizobium japonicum, Brucella melitensis, Escherichia coli, Euglena gracilis, Mesorhizobium loti,  Nostoc sp. PCC7120 , Rhizobium meliloti, Rhodospirillum rubrum, Ruta graveolens, Rhodopseudomonas palustris, Salmonella typhimurium, Serratia marcescens, Sulfolobus solfataricus, Thermobifida fusca,  sorghum, soybean, rice, cotton, wheat, tobacco, and  Zea mays.    
     
     
         30 . An animal feed or human food made by the method of  claim 27 .  
     
     
         31 . An animal feed or human food comprising the DNA construct of  claim 10 .  
     
     
         32 . A transgenic plant made by the method of  claim 27 .  
     
     
         33 . A transgenic plant comprising the DNA construct of  claim 10.

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