US2006057723A1PendingUtilityA1

Transgenic plants containing molecular decoys that alter protein content therein

67
Individually held — no corporate assignee on recordPriority: Aug 30, 2000Filed: Nov 10, 2004Published: Mar 16, 2006
Est. expiryAug 30, 2020(expired)· nominal 20-yr term from priority
Inventors:Mark ConklingYan Li
C12N 15/8223C12N 15/8222C12N 15/8243C12N 15/8257C07K 14/415C12N 15/8251C12N 15/8227C12N 9/1077C12N 15/8216C12N 15/11
67
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Claims

Abstract

The present application describes isolated nucleic acids that contain a Nic gene product responsive element, and the use thereof in methods of producing transgenic tobacco plants having reduced levels of nicotine and/or TSNA therein, as well as other plants or host cells that contain altered levels of a protein of interest therein due to inclusion of a cis-acting decoy element therein.

Claims

exact text as granted — not AI-modified
1 - 10 . (canceled)  
     
     
         11 . A method of making a transgenic tobacco plant having a reduced amount of nicotine, comprising: 
 introducing into at least one tobacco plant cell isolated nucleic acids that are Nic gene product responsive elements, wherein said Nic gene product responsive elements are selected from the group consisting of:    (a) an isolated nucleic acid sequence of SEQ ID NO: 1 or an active fragment thereof wherein said fragment comprises 20 to 455 consecutive nucleotides of SEQ ID NO: 1;    (b) a nucleic acid sequence which is at least 95% identical to the nucleic acid sequence of (a); and    (c) an isolated nucleic acid that hybridizes under stringent conditions to the complement of SEQ ID NO:1 and is responsive to a Nic gene product;    whereby said introduction produces at least one transformed tobacco plant cell, and    regenerating said transformed tobacco plant cell into said transgenic tobacco plant, which has a reduced amount of nicotine as compared to a non-transgenic tobacco plant of the same variety grown under similar conditions.    
     
     
         12 . The method of  claim 11 , further comprising: 
 collecting tobacco leaves from said tobacco plant, wherein said tobacco leaves have a reduced amount of nicotine as compared to the amount of nicotine that would be present in said tobacco plant in the absence of said nucleic acid.    
     
     
         13 . The method according to  claim 11 , further comprising: 
 collecting tobacco seed from said tobacco plant, wherein said tobacco seed comprises said nucleic acid in a copy number sufficient to reduce the amount of nicotine in a tobacco plant produced from said seed as compared to the amount of nicotine that would be present in the absence of said nucleic acid.    
     
     
         14 . (canceled)  
     
     
         15 . The method according to  claim 11 , wherein said Nic gene product responsive element consists of the sequence given herein as SEQ ID NO:1.  
     
     
         16 . The method according to  claim 11 , wherein said nucleic acid is contained within a recombinant nucleic acid construct, and said recombinant nucleic acid construct does not contain a NtQPT1 coding sequence, and said recombinant construct is linear.  
     
     
         17 . The method according to  claim 16 , wherein said recombinant nucleic acid construct is circular.  
     
     
         18 . The method according to  claim 11 , wherein said nucleic acid is a DNA.  
     
     
         19 . The method according to  claim 11 , wherein said introducing step comprises ballistic transformation.  
     
     
         20 . The method according to  claim 11 , wherein said introducing step comprises  Agrobacterium  transformation.  
     
     
         21 . A tobacco plant produced by the method of  claim 11 .  
     
     
         22 . A tobacco leaf collected from a the tobacco plant of  claim 21 .  
     
     
         23 . A tobacco seed collected from the tobacco plant of  claim 21 , wherein said seed comprises a nucleic acid consisting essentially of a Nic gene product responsive element.  
     
     
         24 . A tobacco plant having a reduced amount of nicotine therein, wherein said plant comprises cells comprising an exogenous nucleic acid, wherein said exogenous nucleic acid consists essentially of a Nic gene product responsive element; said exogenous nucleic acid contained in said cells in a copy number sufficient to reduce the amount of nicotine in said tobacco plant as compared to the amount of nicotine that would be present in said plant in the absence of said exogenous nucleic acid.  
     
     
         25 . The tobacco plant according to  claim 24 , wherein said Nic gene product responsive element is selected from the group consisting of: 
 (a) the isolated nucleic acids sequence of SEQ ID NO: 1 or an active fragment thereof, wherein said fragment comprises 20 to 455 consecutive nucleotides of SEQ ID NO: 1;    (b) a nucleic acid sequence which is at least 95% identical to the nucleic acid sequence of (a); and    (c) an isolated nucleic acids that hybridizes under stringent conditions to the complement of SEQ ID NO:1 and is responsive to a Nic gene product.    
     
     
         26 . The tobacco plant according to  claim 24 , wherein said Nic gene product responsive element consists essentially of the sequence given herein as SEQ ID NO:1.  
     
     
         27 . The tobacco plant according to  claim 24 , wherein said exogenous nucleic acid is contained within a recombinant nucleic acid construct, wherein said recombinant nucleic acid construct does not contain a NtQPT1 coding sequence, and said recombinant construct is linear.  
     
     
         28 . The tobacco plant according to  claim 27 , wherein said recombinant nucleic acid construct is circular.  
     
     
         29 . The tobacco plant according to  claim 24 , wherein said exogenous nucleic acid is DNA.  
     
     
         30 . A tobacco leaf collected from the tobacco plant of  claim 24 .  
     
     
         31 . A tobacco seed that germinates into the tobacco plant of  claim 24 .  
     
     
         32 . A tobacco seed collected from the tobacco plant of  claim 24 , wherein said seed comprises cells comprising an exogenous nucleic acid consisting essentially of a Nic gene product responsive element.  
     
     
         33 - 73 . (canceled)  
     
     
         74 . An isolated nucleic acid comprising the isolated nucleic acid sequence of SEQ ID NO: 1 operably linked to a Nic gene product.  
     
     
         75 . The isolated nucleic acid of  claim 74 , wherein said nucleic acid is DNA.  
     
     
         76 . A recombinant nucleic acid construct comprising the isolated nucleic acid of  claim 75 , wherein said recombinant nucleic acid construct does not contain a NtQPT1 coding sequence.  
     
     
         77 . The recombinant nucleic acid construct according to  claim 76 , wherein said recombinant nucleic acid construct is linear.  
     
     
         78 . The recombinant nucleic acid construct according to  claim 76 , wherein said recombinant nucleic acid construct is circular.  
     
     
         79 . A microparticle coated with the isolated nucleic acid construct according to  claim 76 , wherein said microparticle is suitable for transformation of a plant cell.  
     
     
         80 . The recombinant nucleic acid construct according to  claim 76 , wherein said recombinant nucleic acid construct is an  Agrobacterium  vector.  
     
     
         81 . A plant cell comprising the isolated nucleic acid according to  claim 74 .  
     
     
         82 . A transgenic tobacco plant comprising the isolated nucleic acid of claim  1 .  
     
     
         83 . The isolated nucleic acid of claim  1 , wherein the Nic gene product is selected from the group consisting of Nic1, Nic2 and a combination of Nic1 and Nic2.  
     
     
         84 . The isolated nucleic acid of claim  1 , wherein the nucleic acid sequence is at least 95% identical to SEQ ID NO: 1 and is operably linked to a Nic gene product.  
     
     
         85 . An isolated nucleic acid comprising the isolated nucleic acid sequence of SEQ ID NO: 1, further comprising a recombinant nucleic acid construct, wherein said recombinant nucleic acid construct does not contain a NtQPT1 coding sequence.  
     
     
         86 . The recombinant nucleic acid construct according to  claim 85 , wherein said recombinant nucleic acid construct is linear.  
     
     
         87 . The recombinant nucleic acid construct according to  claim 85 , wherein said recombinant nucleic acid construct is circular.  
     
     
         88 . A microparticle coated with the isolated nucleic acid construct according to  claim 85 , wherein said microparticle is suitable for transformation of a plant cell.  
     
     
         89 . The recombinant nucleic acid construct according to  claim 85 , wherein said recombinant nucleic acid construct is an  Agrobacterium  vector.  
     
     
         90 . A plant cell comprising an isolated nucleic acid sequence of SEQ ID NO: 1.  
     
     
         91 . A transgenic tobacco plant comprising an isolated nucleic acid sequence of SEQ ID NO: 1.

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