Patent ID: 6228583
Filing Date: 2001-05-08
Classification: A01K,C07K,C12N,C12Q,G01N

Abstract:
A method of identifying an agent which inhibits the replication or accumulation of ribosomal DNA circles in a yeast cell, comprising the steps of:a) transforming a yeast cell deficient in a gene encoding a protein necessary for growth with a plasmid comprising a ribosomal DNA autonomously replicating sequence and a marker gene encoding the protein necessary for growth, to produce a transformed yeast cell;b) identifying a transformed yeast cell in which the plasmid has stably integrated into the ribosomal DNA genome and a transformed yeast cell in which the plasmid has not integrated into the ribosomal DNA genome;c) incubating the transformed yeast cell in which the plasmid has stably integrated into the ribosomal DNA genome and the transformed yeast cells in which the plasmid has not integrated into the ribosomal DNA genome with the agent under conditions suitable for growth, to produce a test yeast cell in which the plasmid has stably integrated into the ribosomal DNA genome and a test yeast cell in which the plasmid has not integrated into the ribosomal DNA genome;d) assessing growth of the test yeast cell in which the plasmid has stably integrated into the ribosomal DNA genome and a test yeast cell in which the plasmid has not integrated into the genome; ande) comparing the growth of the test yeast cell in which the plasmid has stably integrated into the ribosomal DNA genome with the growth of the test yeast cell in which the plasmid has not integrated into the ribosomal DNA genome,wherein reduced growth of the test yeast cell in which the plasmid has not integrated into the ribosomal DNA genome compared to the growth of the test yeast cell in which the plasiid has stably integrated into the ribosomal DNA genome indicates that the agent inhibits the replication or accumulation of ribosomal DNA circles in a veast cell.