Matrix attachment regions

Abstract

Matrix attachment regions isolated from a higher plants, and DNA constructs and vectors containing such matrix attachment regions, are described. A method of identifying matrix attachment regions is provided.

That which is claimed is:  
     
         1 . An isolated DNA molecule having a nucleotide sequence selected from the group consisting of: 
 (a) SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11and SEQ ID NO:13; and    (b) sequences that hybridize to isolated DNA of (a) above under conditions represented by a wash stringency of 0.3M NaCl, 0.03M sodium citrate, and 0.1% SDS at 60° C., and which encode a matrix attachment region.    
     
     
         2 . A DNA construct comprising: 
 (a) a transcription initiation region and a structural gene positioned downstream from said transcription initiation region and operatively associated therewith; and    (b) a matrix attachment region according to  claim 1  positioned either 5′ to said transcription initiation region or 3′ to said structural gene.    
     
     
         3 . A DNA construct according to  claim 2 , wherein said matrix attachment region is 5′ to said transcription initiation region.  
     
     
         4 . A DNA construct according to  claim 2 , wherein said matrix attachment region is 3′ to said structural gene.  
     
     
         5 . A DNA construct according to  claim 2 , further comprising a second matrix attachment region that differs in sequence from said matrix attachment region according to  claim 1 .  
     
     
         6 . A DNA construct comprising: 
 (a) a transcription initiation region and a structural gene positioned downstream from said transcription initiation region and operatively associated therewith;    (b) a matrix attachment region according to  claim 1  positioned either 5′ to said transcription initiation region or 3′ to said structural gene; and    (c) a second matrix attachment region according to  claim 1 , wherein said second matrix attachment region is positioned either 5′ to said transcription initiation region or 3′ to said structural gene.    
     
     
         7 . A DNA construct according to  claim 2 , further comprising a termination sequence positioned downstream from said structural gene and operatively associated therewith.  
     
     
         8 . A DNA construct according to  claim 2 , wherein said first and said second matrix attachment regions differ in sequence.  
     
     
         9 . A vector comprising a DNA construct according to  claim 2 .  
     
     
         10 . A vector according to  claim 9 , wherein said vector is selected from the group consisting of plasmids, viruses, and plant transformation vectors.  
     
     
         11 . A host cell containing a DNA construct according to  claim 2 .  
     
     
         12 . A host cell according to  claim 9 , wherein said host cell is an animal cell or a plant cell.  
     
     
         13 . A transgenic plant comprising transformed plant cells, said transformed plant cells containing a DNA construct according to  claim 2 .  
     
     
         14 . A transgenic plant according to  claim 13 , which is a monocot.  
     
     
         15 . A transgenic plant according to  claim 13 , which is a dicot.  
     
     
         16 . A transgenic plant according to  claim 13 , which plant is a dicot selected from the group consisting of tobacco, potato, soybean, peanuts, cotton, and vegetable crops.  
     
     
         17 . A DNA construct comprising, in the 5′ to 3′ direction, a transcription initiation region, a structural gene positioned downstream from said transcription initiation region and operatively associated therewith, and a matrix attachment region positioned either 5′ to said transcription initiation region or 3′ to said structural gene, wherein said matrix attachment region has a sequence selected from SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:13; 
 said DNA construct carried by a plant transformation vector.  
 
     
     
         18 . A DNA construct according to  claim 17 , further comprising a second matrix attachment region that differs in sequence from said matrix attachment region.  
     
     
         19 . A recombinant tobacco plant comprising transformed tobacco plant cells, said transformed tobacco plant cells containing a heterologous DNA construct comprising, in the 5′ to 3′ direction, a transcription initiation region functional in plant cells, a structural gene positioned downstream from said transcription initiation region and operatively associated therewith, and a matrix attachment region positioned either 5′ to said transcription initiation region or 3′ to said structural gene, 
 wherein said matrix attachment region has a sequence selected from SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11 and SEQ ID NO:13.  
 
     
     
         20 . A method of identifying matrix attachment regions in a DNA molecule of known nucleotide sequence, comprising identifying a sequence section of at least twenty contiguous nucleotides that is at least 90% A or T nucleotides, wherein the presence of such a sequence section indicates a MAR encompassing said sequence section.  
     
     
         21 . A method according to claim  20 , further comprising preparing a MAR molecule of at least about 300 nucleotides, said MAR having a sequence which is a contiguous fragment of said DNA molecule sequence and which encompasses said identified sequence section of at least twenty contiguous nucleotides that is at least 90% A or T nucleotides.