TY - JOUR
T1 - Chromosome-shuffling technique for selected chromosomal segments in Saccharomyces cerevisiae
AU - Sugiyama, Minetaka
AU - Yamamoto, Eishi
AU - Mukai, Yukio
AU - Kaneko, Yoshinobu
AU - Nishizawa, Masafumi
AU - Harashima, Satoshi
N1 - Funding Information:
Acknowledgements Part of this work was financially supported by the Ministry of Education, Culture, Sports, Science and Technology, a Grant-in-Aid for Scientific Research B, 15380064, 2003 to 2005 to S.H. and carried out as part of the Project for the Development of a Technological Infrastructure for Industrial Bioprocesses on R & D of New Industrial Science and Technology Frontiers by the Ministry of Economy, Trade and Industry (METI), and entrusted by the New Energy and Industrial Technology Development Organization (NEDO).
PY - 2006/10
Y1 - 2006/10
N2 - We describe a novel chromosome engineering technique for shuffling selected regions of chromosomes from two strains in Saccharomyces cerevisiae: The technique starts with the construction of MAT a and MATα strains in which a particular chromosome is split at exactly the same site in both strains such that the split chromosomes generated are marked with different markers. The two strains are then crossed, and the resultant diploid is cultivated in nutrient medium to induce loss of the split chromosome originating from either of the strains. We predicted that some of these clones that are hemizygous for the split chromosome would spontaneously restore a homozygous configuration of the split chromosome during cultivation. We verified this prediction by tetrad analysis and quantitative Southern hybridization analysis, indicating that it is possible to create diploid hybrids in which a selected region of a chromosome from one strain is replaced by the corresponding chromosomal region from another strain. We also found that some chromosomal segments maintain a hemizygous state. This novel technique, which we call 'chromosome shuffling', could provide a new tool to analyze phenotypic alterations caused by the replacement or hemizygosity of a selected chromosomal region in not only laboratory but also industrial strains of S. cerevisiae.
AB - We describe a novel chromosome engineering technique for shuffling selected regions of chromosomes from two strains in Saccharomyces cerevisiae: The technique starts with the construction of MAT a and MATα strains in which a particular chromosome is split at exactly the same site in both strains such that the split chromosomes generated are marked with different markers. The two strains are then crossed, and the resultant diploid is cultivated in nutrient medium to induce loss of the split chromosome originating from either of the strains. We predicted that some of these clones that are hemizygous for the split chromosome would spontaneously restore a homozygous configuration of the split chromosome during cultivation. We verified this prediction by tetrad analysis and quantitative Southern hybridization analysis, indicating that it is possible to create diploid hybrids in which a selected region of a chromosome from one strain is replaced by the corresponding chromosomal region from another strain. We also found that some chromosomal segments maintain a hemizygous state. This novel technique, which we call 'chromosome shuffling', could provide a new tool to analyze phenotypic alterations caused by the replacement or hemizygosity of a selected chromosomal region in not only laboratory but also industrial strains of S. cerevisiae.
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U2 - 10.1007/s00253-006-0342-5
DO - 10.1007/s00253-006-0342-5
M3 - Article
C2 - 16505990
AN - SCOPUS:33749186030
SN - 0175-7598
VL - 72
SP - 947
EP - 952
JO - Applied Microbiology and Biotechnology
JF - Applied Microbiology and Biotechnology
IS - 5
ER -