원문정보
초록
영어
Versatile technology for manipulation of chromosome is playing an increasingly important role in functional analysis of genomes. Here we describe a simple but innovative method to split chromosomes in Saccharomyces cerevisiae, which we call PCR-mediated chromosome splitting (PCS). The PCS method combines a streamlined procedure (two-step PCR and one transformation per splitting event). Moreover, this method was applied to analyze yeast rDNA-concerned cellular physiology. The rDNA cluster in S. cerevisiae is located in chromosome XII and consists of ca.150 tandemly repeated copies of a 9.1-kb rDNA unit. To explore the biological significance of this specific chromosomal context, chromosome XII was split at both sides of the rDNA cluster by using PCS method and strain harboring chromosome XII consisting of rDNA chromosome (1,500-kb rDNA cluster only) was created. In the strain harboring the rDNA chromosome, the copy number and size of rDNA cluster were decreased and life span also was shorted. These observations suggest that the context of chromosome XII is important for rDNA function and the splitting method is useful tool to study the functional analysis of eukaryotic genome.