Artificial chromosomes, uses thereof and methods for preparing artificial chromosomes

Abstract

Methods for amplification of nucleic acids in cells are provided. Also provided are nucleic acid molecules for amplification.

What is claimed:  
     
         1 . A method for amplifying nucleic acid, comprising: 
 introducing a nucleic acid molecule into a plant cell, wherein the nucleic acid molecule include a sequence of nucleotides that targets it to an amplifiable region of a chromosome in the cell;    growing the cell; and    identifying from among the resulting cells those that include a chromosome with a portion that has undergone amplification.    
     
     
         2 . The method of  claim 1 , wherein the targeting sequence of nucleotide(s) is selected from among those that target the molecule to the pericentric heterochromatic region of a chromosome.  
     
     
         3 . The method of  claim 1 , wherein the targeting sequence comprises rDNA.  
     
     
         4 . The method of  claim 1 , wherein the targeting sequence comprises an origin of replication or an amplification promoting sequence (APS).  
     
     
         5 . The method of  claim 1 , wherein the plant is tobacco, rice, maize, rye, soybean,  Brassica napus,  cotton, lettuce, potato, tomato or arabidopsis.  
     
     
         6 . A method for amplifying a nucleic acid, comprising: 
 introducing nucleic acid fragment comprising sequences of nucleotides targeted to an amplifiable region of a chromosome into a plant cell under conditions whereby the fragment integrates into the chromosome.    
     
     
         7 . The method of  claim 6 , further comprising replicating the cell.  
     
     
         8 . The method of  claim 6 , wherein the targeting sequences of nucleotides are selected from among those that target the molecule to the pericentric heterochromatic region of a chromosome.  
     
     
         9 . The method of  claim 6 , wherein the targeting sequences comprise rDNA.  
     
     
         10 . The method of  claim 6 , wherein the targeting sequences comprise an origin of replication or an amplification promoting sequence (APS).  
     
     
         11 . The method of  claim 6 , wherein the plant is tobacco, rice, maize, rye, soybean,  Brassica napus,  cotton, lettuce, potato, tomato or arabidopsis.  
     
     
         12 . A method for amplifying a nucleic acid, comprising: 
 introducing nucleic acid fragment that comprises rDNA into a plant cell under conditions that produce cells that have incorporated the DNA fragment or a portion thereof that comprises the rDNA into a chromosome of the cell.    
     
     
         13 . The method of  claim 12 , wherein the plant is tobacco, rice, maize, rye, soybean,  Brassica napus,  cotton, lettuce, potato, tomato or arabidopsis.  
     
     
         14 . A method for amplifying a nucleic acid, comprising: 
 introducing nucleic acid fragment that comprises an origin of replication or an amplification promoting sequence into a plant cell under conditions to produce cells that have incorporated the DNA fragment or a portion thereof that comprises the origin of replication or an amplification promoting sequence into a chromosome of the cell.    
     
     
         15 . The method of  claim 14 , wherein the plant is tobacco, rice, maize, rye, soybean,  Brassica napus,  cotton, lettuce, potato, tomato or arabidopsis.  
     
     
         16 . A nucleic acid molecule, comprising: 
 nucleic acid encoding a gene product or gene products;    a selectable marker; and    sequences of nucleotides targeted to an amplifiable region of a chromosome in a cell.    
     
     
         17 . The nucleic acid molecule of  claim 16 , wherein the targeting sequences of nucleotides are selected from among those that target the molecule to the pericentric heterochromatic region of a chromosome.  
     
     
         18 . The nucleic acid molecule of  claim 16 , wherein the targeting sequences comprise rDNA.  
     
     
         19 . The nucleic acid molecule of  claim 16 , wherein the targeting sequences comprise an origin of replication or an amplification promoting sequence (APS).  
     
     
         20 . The nucleic acid molecule of  claim 16 , wherein the gene products encode a biosynthetic pathway.  
     
     
         21 . The nucleic acid molecule of  claim 16  that is a plasmid.  
     
     
         22 . The nucleic acid molecule of  claim 16 , wherein the cell is a plant cell.  
     
     
         23 . The nucleic acid molecule of  claim 16 , wherein the plant is tobacco, rice, maize, rye, soybean,  Brassica napus,  cotton, lettuce, potato, tomato or arabidopsis.