Artificial chromosomes, uses thereof and methods for preparing artificial chromosomes

Abstract

Methods for preparing cell lines that contain artificial chromosomes, methods for preparation of artificial chromosomes, methods for purification of artificial chromosomes, methods for targeted insertion of heterologous DNA into artificial chromosomes, methods for amplification of nucleic acids and methods for delivery of the chromosomes to selected cells and tissues are provided. Also provided are cell lines for use in the methods, and cell lines and chromosomes produced by the methods. Methods for use of the artificial chromosomes are also provided.

1 . A plant cell produced by a method comprising:
 introducing a DNA fragment into a plant cell, wherein the DNA fragment comprises a selectable marker;   growing the cell under selective conditions to produce cells that have incorporated the DNA fragment into their genomic DNA, whereby amplification of pericentric heterochromatin occurs; and   selecting a plant cell that comprises the amplified heterochromatin.   
     
     
         2 . The plant cell of  claim 1 , wherein the amplified heterochromatin is in a sausage chromosome. 
     
     
         3 . The plant cell of  claim 1  that is a plant protoplast.