Plant regulatory sequences for selective control of gene expression

Abstract

Promoters from male reproductive tissues were isolated from corn ( Zea mays ). These promoters can be used in plants to regulate transcription of target genes including genes for control of fertility, insect or pathogen tolerance, herbicide tolerance or any gene of interest.

What is claimed is:  
     
         1 . An isolated nucleic acid comprising a sequence selected from the group consisting of SEQ ID NOS:79-98, or a fragment, region, or cis element of said sequence thereof, said isolated nucleic acid being capable of regulating transcription of an operably linked DNA sequence.  
     
     
         2 . The isolated nucleic acid of  claim 1  wherein the isolated nucleic acid is a promoter.  
     
     
         3 . The isolated nucleic acid of  claim 2  wherein the promoter is a hybrid promoter.  
     
     
         4 . The isolated nucleic acid of  claim 3  wherein said isolated nucleic acid confers enhanced expression of operably linked genes in male reproductive tissues.  
     
     
         5 . The isolated nucleic acid of  claim 4  wherein said isolated nucleic acid confers enhanced expression of operably linked genes in anthers.  
     
     
         6 . The isolated nucleic acid of  claim 5  wherein said isolated nucleic acid confers enhanced expression of operably linked genes in wheat anthers.  
     
     
         7 . The isolated nucleic acid of  claim 4  further comprising a minimal promoter.  
     
     
         8 . The isolated nucleic acid of  claim 7  wherein the minimal promoter is selected from the group consisting of a minimal CaMV and a rice actin promoter.  
     
     
         9 . The isolated nucleic acid of  claim 8  wherein the minimal promoter is a minimal CaMV 35S promoter.  
     
     
         10 . A promoter comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOS: 79-98 and fragments thereof.  
     
     
         11 . The promoter of  claim 10  wherein said promoter confers enhanced expression of operably linked genes in male reproductive tissues.  
     
     
         12 . The promoter of  claim 11  wherein said promoter confers enhanced expression of operably linked genes in anthers.  
     
     
         13 . The promoter of  claim 12  wherein said promoter confers enhanced expression of operably linked genes in wheat anthers.  
     
     
         14 . A cell comprising a DNA construct comprising an isolated nucleic acid sequence selected from the group consisting of SEQ ID NOS:79-98 or a fragment, region, or cis element of said sequence thereof, and operably linked to said nucleic acid sequence, a transcribable DNA sequence and a 3′ non-translated region.  
     
     
         15 . A transgenic plant comprising a DNA construct comprising an isolated nucleic acid sequence selected from the group consisting of SEQ ID NOS:79-98 or a fragment, region, or cis element of said sequence thereof, and operably linked to said nucleic acid sequence, a transcribable DNA sequence and a 3′ non-translated region.  
     
     
         16 . A method of regulating transcription of a DNA sequence comprising operably linking the DNA sequence to a promoter comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOS:79-98.  
     
     
         17 . The method of  claim 16  comprising operably linking the DNA sequence to a hybrid promoter comprising the nucleic acid sequence selected from the group consisting of SEQ ID NOS:79-98.  
     
     
         18 . The method of  claim 16  wherein operably linking the nucleic acid sequence selected from the group consisting of SEQ ID NOS:79-98 or fragment thereof to the promoter confers enhanced expression of operably linked genes in male reproductive tissues.  
     
     
         19 . The method of  claim 18  wherein said male reproductive tissues comprise monocot or dicot male reproductive tissues.  
     
     
         20 . The method of  claim 19  wherein said male reproductive tissues comprise anthers.  
     
     
         21 . The method of  claim 20  wherein said male reproductive tissues comprise wheat anthers.  
     
     
         22 . The method of  claim 16  comprising operably linking a minimal promoter to the nucleic acid sequences selected from the group consisting of SEQ ID NOS:79-98 or fragment, region, or cis element thereof.  
     
     
         23 . A method of making a transgenic plant comprising introducing into a cell of a plant a DNA construct comprising: (i) a promoter comprising a nucleic acid sequence selected from the group consisting of SEQ ID NOS:79-98 or a fragment, region or cis element thereof, and, operably linked to the promoter, (ii) a transcribable DNA sequence and (iii) a 3′ non-translated region.  
     
     
         24 . A method of isolating at least two 5′ regulatory sequences that confer enhanced expression of operably linked genes in male reproductive tissues from a plant comprising: 
 (i) evaluating a collection of nucleic acid sequences of ESTs derived from at least one cDNA library prepared from a plant cell type of interest;  
 (ii) comparing EST sequences from at least one target plant cDNA library and at least one non-target cDNA libraries of ESTs from a different plant cell type;  
 (iii) subtracting common EST sequences found in both target and non-target libraries;  
 (iv) designing gene specific primers from the remaining EST sequences after said subtraction; and  
 (v) isolating the corresponding 5′ flanking and regulatory sequences from a genomic library prepared from the target plant comprising the use of said primers.  
 
     
     
         25 . The method of  claim 24  wherein said male reproductive tissues are from monocot or dicot plants.  
     
     
         26 . The method of  claim 25  wherein said male reproductive tissues comprise anthers.  
     
     
         27 . The method of  claim 26  wherein said male reproductive tissues comprise wheat anthers.