Methods and compositions for protein production in tobacco plants with reduced nicotine

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.

1 - 73 . (canceled)  
     
     
         74 . A transgenic tobacco plant having a reduced amount of nicotine therein, wherein said transgenic tobacco plant comprises cells comprising an exogenous nucleic acid sequence that binds a Nic gene product, wherein said exogenous nucleic acid sequence is 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 and binds a Nic gene product;    (b) an isolated nucleic acid sequence which is at least 80% identical to the nucleic acid sequence of (a) and binds a Nic gene product; and    (c) an isolated nucleic acid that hybridizes under stringent conditions to the complement of the nucleic acid sequence of (a) and binds a Nic gene product, and wherein said exogenous nucleic acid is present 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 sequence,    and wherein the transgenic tobacco plant further comprises a transgene encoding a heterologous protein of interest.    
     
     
         75 . The transgenic tobacco plant of claim  1 , wherein said exogenous nucleic acid sequence consists essentially of the nucleotide sequence of SEQ ID NO:1.  
     
     
         76 . The transgenic tobacco plant of claim  1 , wherein said exogenous nucleic acid sequence is contained within a recombinant nucleic acid construct, wherein said recombinant nucleic acid construct does not contain a NtQPT1 coding sequence.  
     
     
         77 . The transgenic tobacco plant of claim  1 , wherein said isolated nucleic acid sequence of SEQ ID NO:1 is a fragment comprising 30-400 consecutive nucleotides of SEQ ID NO:1 and binds a Nic gene product.  
     
     
         78 . The transgenic tobacco plant of claim  1 , wherein said isolated nucleic acid sequence of SEQ ID NO:1 is a fragment comprising 50-350 consecutive nucleotides of SEQ ID NO:1 and binds a Nic gene product.  
     
     
         79 . The transgenic tobacco plant of claim  1 , wherein said isolated nucleic acid sequence of SEQ ID NO:1 is a fragment comprising 100-300 consecutive nucleotides of SEQ ID NO:1 and binds a Nic gene product.  
     
     
         80 . The transgenic tobacco plant of claim  1 , wherein said isolated nucleic acid sequence of SEQ ID NO:1 is a fragment comprising 200-400 consecutive nucleotides of SEQ ID NO:1 and binds a Nic gene product.  
     
     
         81 . The transgenic tobacco plant of claim  1 , wherein said isolated nucleic acid sequence of SEQ ID NO:1 is a fragment of at least 20 consecutive nucleotides of SEQ ID NO:1 and binds a Nic gene product.  
     
     
         82 . The transgenic tobacco plant of claim  1 , wherein said isolated nucleic acid sequence of SEQ ID NO:1 is a fragment of at least 40 consecutive nucleotides of SEQ ID NO:1 and binds a Nic gene product.  
     
     
         83 . The transgenic tobacco plant of claim  1 , wherein said isolated nucleic acid sequence of SEQ ID NO:1 is a fragment of at least 60 consecutive nucleotides of SEQ ID NO:1 and binds a Nic gene product.  
     
     
         84 . A method of producing a heterologous protein of interest in a plant, comprising: 
 a) growing the transgenic tobacco plant of claim  1 ; and    b) collecting the heterologous protein of interest from said transgenic plant.    
     
     
         85 . A method of producing a heterologous protein of interest in a plant, comprising: 
 a) growing the transgenic tobacco plant of claim  2 ; and    b) collecting the heterologous protein of interest from said transgenic plant.    
     
     
         86 . A method of producing a heterologous protein of interest in a plant, comprising: 
 a) growing the transgenic tobacco plant of claim  3 ; and    b) collecting the heterologous protein of interest from said transgenic plant.    
     
     
         87 . A method of producing a heterologous protein of interest in a plant, comprising: 
 a) growing the transgenic tobacco plant of claim  9 ; and    b) collecting the heterologous protein of interest from said transgenic plant.    
     
     
         88 . A method of producing a heterologous protein of interest in a plant, comprising: 
 a) growing the transgenic tobacco plant of claim  10 ; and    b) collecting the heterologous protein of interest from said transgenic plant.    
     
     
         89 . A method of producing a heterologous protein of interest in a plant, comprising: 
 a) growing the transgenic tobacco plant of claim  11 ; and    b) collecting the heterologous protein of interest from said transgenic plant.    
     
     
         90 . The transgenic tobacco plant of claim  1 , wherein the heterologous protein of interest is a  Bacillus thuringiensis  insecticidal protein.  
     
     
         91 . The method of claim  12 , wherein the heterologous protein of interest is a  Bacillus thuringiensis  insecticidal protein.