Plant centromere compositions

Abstract

The present invention provides for the nucleic acid sequences of plant centromeres. This will permit construction of stably inherited recombinant DNA constructs and minichromosomes which can serve as vectors for the construction of transgenic plant and animal cells.

1 - 96 . (canceled) 
     
     
         97 . A minichromosome comprising a plant centromere, wherein the plant centromere comprises n copies of a repeated nucleotide sequence obtained from total genomic DNA of the plant, and wherein n is at least 2. 
     
     
         98 - 99 . (canceled) 
     
     
         100 . The minichromosome of  claim 97 , further comprising a structural gene. 
     
     
         101 . The minichromosome of  claim 100 , wherein the structural gene is selected from the group consisting of a selectable or screenable marker gene, an antibiotic resistance gene, a ligand gene, an enzyme gene, a herbicide resistance gene, a nitrogen fixation gene, a plant pathogen defense gene, a plant stress-induced gene, a toxin gene, a receptor gene, a gene encoding an enzyme, a gene encoding an antibody, a gene encoding an antigen for a vaccine, a transcription factor, a cytoskeletal protein, a DNA-binding protein, a protease, an endonuclease, a lipid, a seed storage gene, an interleukin gene, a clotting factor gene, a cytokine gene, a growth factor gene and a biosynthetic gene for producing pharmaceutically active proteins, small molecules with medicinal properties, chemicals with industrial utility, nutraceuticals, carbohydrates, RNAs, lipids, fuels, dyes, pigments, vitamins, scents, flavors, vaccines, antibodies, and hormones. 
     
     
         102 . The minichromosome of  claim 100 , wherein the minichromosome is capable of expressing the desired structural gene. 
     
     
         103 . (canceled) 
     
     
         104 . A transgenic crop comprising a crop cell transformed with a minichromosome that comprises a plant centromere, the centromere comprises n copies of a repeated nucleotide sequence, and wherein n is at least 2. 
     
     
         105 . The transgenic crop of  claim 104 , wherein the minichromosome further comprises at least one structural gene. 
     
     
         106 . The transgenic crop of  claim 105 , wherein the structural gene is selected from the group consisting of a selectable or screenable marker gene, an antibiotic resistance gene, a ligand gene, an enzyme gene, a herbicide resistance gene, a nitrogen fixation gene, a plant pathogen defense gene, a plant stress-induced gene, a toxin gene, a receptor gene, a gene encoding an enzyme, a gene encoding an antibody, a gene encoding an antigen for a vaccine, a transcription factor, a cytoskeletal protein, a DNA-binding protein, a protease, an endonuclease, a lipid, a seed storage gene, an interleukin gene, a clotting factor gene, a cytokine gene, a growth factor gene and a biosynthetic gene for producing pharmaceutically active proteins, small molecules with medicinal properties, chemicals with industrial utility, nutraceuticals, carbohydrates, RNAs, lipids, fuels, dyes, pigments, vitamins, scents, flavors, vaccines, antibodies, and hormones. 
     
     
         107 . The transgenic crop of  claim 104 , wherein the crop is a dicotyledonous plant. 
     
     
         108 . The transgenic crop of  claim 107 , wherein the dicotyledonous plant is selected from the group consisting of tobacco, tomato, potato, sugar beet, pea, carrot, cauliflower, broccoli, soybean, canola, sunflower, alfalfa, and cotton. 
     
     
         109 . The transgenic crop of  claim 104 , wherein the crop is a monocotyledonous plant. 
     
     
         110 . The transgenic crop of  claim 109 , wherein the monocotyledonous plant is selected from the group consisting of wheat, maize, rye, rice, turfgrass, oat, barley, sorghum, millet, and sugarcane. 
     
     
         111 . (canceled) 
     
     
         112 . A method for preparing a transgenic crop tissue, the method comprising the steps of contacting a starting crop tissue with a recombinant DNA construct comprising a plant centromere, the centromere comprises n copies of a repeated nucleotide sequence, wherein n is at least 2, whereby the starting crop tissue is transformed with the recombinant DNA construct. 
     
     
         113 - 127 . (canceled) 
     
     
         128 . A method of preparing a transgenic plant cell comprising contacting a starting plant cell with a minichromosome of  claim 97 . 
     
     
         129 . The minichromosome of  claim 97 , wherein the minichromosome is circular. 
     
     
         130 . The minichromosome of  claim 97 , wherein the centromere comprises at least five copies of the repeated nucleotide sequence. 
     
     
         131 . The minichromosome of  claim 97 , wherein the centromere has less than 1000 copies of the repeated nucleotide sequence. 
     
     
         132 . The isolated minichromosome of  claim 100 , wherein at least one structural gene is operably linked to a heterologous regulatory sequence functional in plant cells. 
     
     
         133 . The isolated minichromosome of  claim 100 , wherein the minichromosome comprises at least five structural genes. 
     
     
         134 . The isolated minichromosome of  claim 100 , wherein the minichromosome comprises at least ten structural genes. 
     
     
         135 . The isolated minichromosome of  claim 100 , wherein at least one structural gene is operably linked to a plant promoter.