Plant chromosome compositions and methods

Abstract

The present invention provides for the identification and cloning of functional plant centromeres in Arabidopsis . This will permit construction of stably inherited minichromosomes which can serve as vectors for the construction of transgenic plant and animal cells. In addition, information on the structure and function of these regions will prove valuable in isolating additional centromeric and centromere related genetic elements and polypeptides from other species.

1 . A recombinant DNA construct comprising a plant centromere.  
     
     
         2 - 8 . (canceled)  
     
     
         9 . The recombinant DNA construct of  claim 1 , which additionally comprises a structural gene.  
     
     
         10 . The recombinant DNA construct of  claim 9 , wherein the structural gene comprises a selectable or screenable marker gene.  
     
     
         11 - 38 . (canceled)  
     
     
         39 . The recombinant DNA construct of  claim 1 , which is capable of being maintained as a chromosome, wherein said chromosome is transmitted in dividing cells.  
     
     
         40 - 91 . (canceled)  
     
     
         92 . The recombinant DNA construct of  claim 1 , comprising n copies of a repeated nucleotide sequence, wherein n is at least 2.  
     
     
         93 - 102 . (canceled)  
     
     
         103 . A minichromosome vector comprising a plant centromere and a telomere sequence.  
     
     
         104 . The minichromosome vector of  claim 103 , comprising an autonomous replicating sequence.  
     
     
         105 . The minichromosome vector of  claim 103 , comprising a second telomere sequence.  
     
     
         106 . The minichromosome vector of  claim 103 , comprising a structural gene.  
     
     
         107 . The minichromosome vector of  claim 103 , further defined as comprising a second structural gene.  
     
     
         108 . (canceled)  
     
     
         109 . A cell transformed with a recombinant DNA construct of  claim 1 .  
     
     
         110 - 127 . (canceled)  
     
     
         128 . A plant comprising the cell of  claim 109 .  
     
     
         129 . A method of preparing a transgenic plant cell comprising contacting a starting plant cell with a recombinant DNA construct of  claim 1 , whereby said starting plant cell is transformed with said recombinant DNA construct.  
     
     
         130 . The method of  claim 129 , wherein said recombinant DNA construct comprises a structural gene.  
     
     
         131 . The method of  claim 130 , wherein the recombinant DNA construct comprises a second structural gene.  
     
     
         132 - 133 . (canceled)  
     
     
         134 . A transgenic plant comprising a minichromosome vector of  claim 103 .  
     
     
         135 - 146 . (canceled)  
     
     
         147 . A method of producing a minichromosome vector of  claim 103  comprising 
 (a) obtaining a first vector and a second vector, wherein said first vector or said second vector comprises a selectable or screenable marker, an origin of replication, a telomere, and a plant centromere, and wherein said first vector and said second vector comprises a site for site-specific recombination; and    (b) contacting said first vector with said second vector to allow site-specific recombination to occur between said site for site-specific recombination on said first vector and said site for site-specific recombination on said second vector to create a minichromosome vector comprising said selectable or screenable marker, said origin of replication, said telomere and said plant centromere.    
     
     
         148 - 163 . (canceled)  
     
     
         164 . A method of screening a candidate centromere sequence for plant centromere activity, said method comprising the steps of 
 (a) obtaining an isolated nucleic acid sequence comprising a candidate centromere sequence;    (b) integratively transforming plant cells with said isolated nucleic acid; and    (c) screening for centromere activity of said candidate centromere sequence.    
     
     
         165 - 231 . (canceled)