Novel constitutive and tissue specific plant promoters and their utilization

Abstract

A recombinant promoter which is functional in plant cells, the recombinant promoter comprises at least a minimal functional plant promoter region derived from SEQ ID NOs: 7 or 8.

What is claimed is:  
     
         1 . A method of isolating a minimal functional plant promoter region, the method comprising 
 connecting upstream to a reporter gene a fragment derived from SEQ ID NO:7; and    testing for a level of expression of the reporter gene in a plant expression system.    
     
     
         2 . The method of  claim 1 , wherein said fragment derived from SEQ ID NO:7 is obtained by PCR amplification, restriction with at least one endonuclease or solid phase DNA synthesis.  
     
     
         3 . The method of  claim 1 , wherein said plant expression system is a plant cell culture, a transgenic plant or a transiently transformed plant.  
     
     
         4 . A method of isolating a minimal functional plant promoter region, the method comprising 
 connecting upstream to a reporter gene a fragment derived from SEQ ID NO:8; and    testing for a level of expression of the reporter gene in a plant expression system.    
     
     
         5 . The method of  claim 4 , wherein said fragment derived from SEQ ID NO:8 is obtained by PCR amplification, restriction with at least one endonuclease or solid phase DNA synthesis.  
     
     
         6 . The method of  claim 4 , wherein said plant expression system is a plant cell culture, a transgenic plant or a transiently transformed plant.  
     
     
         7 . A recombinant promoter which is functional in plant cells, wherein said recombinant promoter comprises at least a minimal functional plant promoter region derived from SEQ ID NO:7.  
     
     
         8 . An expression construct comprising the recombinant promoter of  claim 7  and a transcription terminator functional in plant cells.  
     
     
         9 . The expression construct of  claim 8 , further comprising a structural gene of interest under control of said recombinant promoter.  
     
     
         10 . The expression construct of  claim 8 , further comprising an antisense polynucleotide of interest under control of said recombinant promoter.  
     
     
         11 . A plant cell transformed or transfected with a structural gene of interest under the control of the recombinant promoter of  claim 7 .  
     
     
         12 . A plant cell transformed or transfected with an antisense polynucleotide of interest under the control of the recombinant promoter of  claim 7 .  
     
     
         13 . A plant cell transformed or transfected with the expression construct of  claim 9 .  
     
     
         14 . A method for expressing in plant cells a structural gene of interest, the method comprising transforming or transfecting plant cells with said structural gene of interest under control of the recombinant promoter of  claim 7 .  
     
     
         15 . A method for expressing in plant cells an antisense polynucleotide of interest, the method comprising transforming or transfecting plant cells with said antisense polynucleotide of interest under control of the recombinant promoter of  claim 7 .  
     
     
         16 . The method of  claim 14 , wherein said plant cells are part of a whole plant.  
     
     
         17 . The method of  claim 14 , wherein said plant cells are isolated from a plant.  
     
     
         18 . A method of constructing an expression construct, the method comprising connecting in an operable 5′ to 3′ orientation: 
 the recombinant promoter of  claim 1;   
 a structural gene of interest; and  
 a transcription terminator functional in plant cells.  
 
     
     
         19 . A method of constructing an expression construct, the method comprising connecting in an operable 5′ to 3′ orientation: 
 the recombinant promoter of  claim 1;  and  
 an antisense polynucleotide of interest.  
 
     
     
         20 . A recombinant promoter which is functional in plant cells, wherein said recombinant promoter comprises at least a minimal functional plant promoter region derived from SEQ ID NO:8.  
     
     
         21 . An expression construct comprising the recombinant promoter of  claim 20  and a transcription terminator functional in plant cells.  
     
     
         22 . The expression construct of  claim 21 , further comprising a structural gene of interest under control of said recombinant promoter.  
     
     
         23 . The expression construct of  claim 21 , further comprising an antisense polynucleotide of interest under control of said recombinant promoter.  
     
     
         24 . A plant cell transformed or transfected with a structural gene of interest under the control of the recombinant promoter of  claim 20 .  
     
     
         25 . A plant cell transformed or transfected with an antisense polynucleotide of interest under the control of the recombinant promoter of  claim 20 .  
     
     
         26 . A plant cell transformed or transfected with the expression construct of  claim 22 .  
     
     
         27 . A method for expressing in plant cells a structural gene of interest, the method comprising transforming or transfecting plant cells with said structural gene of interest under control of the recombinant promoter of  claim 20 .  
     
     
         28 . A method for expressing in plant cells an antisense polynucleotide of interest, the method comprising transforming or transfecting plant cells with said antisense polynucleotide of interest under control of the recombinant promoter of  claim 20 .  
     
     
         29 . The method of  claim 27 , wherein said plant cells are part of a whole plant.  
     
     
         30 . The method of  claim 27 , wherein said plant cells are isolated from a plant.  
     
     
         31 . A method of constructing an expression construct, the method comprising connecting in an operable 5′ to 3′ orientation: 
 the recombinant promoter of  claim 20;   
 a structural gene of interest; and  
 a transcription terminator functional in plant cells.  
 
     
     
         32 . A method of constructing an expression construct, the method comprising connecting in an operable 5′ to 3′ orientation: 
 the recombinant promoter of  claim 20;  and  
 an antisense polynucleotide of interest.  
 
     
     
         33 . An isolated polynucleotide comprising a promoter sequence controlling an expression of a ribulose 1,5-bisphosphate carboxylase small subunit, ats3B of a plant.  
     
     
         34 . An isolated polynucleotide comprising a promoter sequence controlling an expression of a photosystem II type I chlorophyll a/b binding protein of a plant.