Construction and Characterization of Single-Gene Chromosomes in Saccharomyces cerevisiae

Based on a previously developed PCR-mediated chromosome splitting method, a genome engineering technique was developed in haploid Saccharomyces cerevisiae for incorporating any desired chromosomal region into a chromosome that carries a single gene. Based on the viability of cells carrying an essential gene in such a construct, close physical proximity of two telomeres and a centromere does not appear to compromise gene function. Spontaneous loss of constructed single-gene chromosomes during vegetative growth was high (0.2-0.4 per cell division), suggesting the possibility of creating novel cells carrying single-gene chromosomes derived from various chromosomal regions in a variety of combinations by exploiting combinatorial loss.