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.
Yamagishi
Kazuo
K
Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita-shi, Osaka 565-0871, Japan.
Sugiyama
Minetaka
M
Kaneko
Yoshinobu
Y
Nishizawa
Masafumi
M
Harashima
Satoshi
S
D015139:Blotting, Southern; D015825:Chromosomes, Fungal; D004587:Electrophoresis, Agar Gel; D005800:Genes, Fungal; D012441:Saccharomyces cerevisiae / Q000235:genetics*