A novel strategy for analysis of gene homologues and segmental genome duplications.
Transformation-associated recombination (TAR) cloning allows selective isolation of a desired chromosomal region or gene from complex genomes. The method exploits a high level of recombination between homologous DNA sequences during transformation in the yeast Saccharomyces cerevisiae. We investigated the effect of nonhomology on the efficiency of gene capture and found that up to 15% DNA divergence did not prevent efficient gene isolation. Such tolerance to DNA divergence greatly expands the potential applications of TAR cloning for comparative genomics. In this study, we were able to use the technique to isolate nonidentical chromosomal duplications and gene homologues.
Noskov
Vladimir N
VN
Laboratory of Biosystems and Cancer, Center for Cancer Research, National Cancer Institute, NIH, Building 37, Room 5032, Bethesda, MD 20892, USA.
Leem
Sun-Hee
SH
Solomon
Greg
G
Mullokandov
Michael
M
Chae
Ji-Youn
JY
Yoon
Young-Ho
YH
Shin
Young-Sun
YS
Kouprina
Natalay
N
Larionov
Vladimir
V
D002503:Centromere / Q000235:genetics; D022202:Chromosomes, Artificial, Bacterial; D002888:Chromosomes, Human, Pair 19; D003001:Cloning, Molecular / Q000379:methods; D020440:Gene Duplication; D016678:Genome; D015894:Genome, Human; D006801:Humans; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics; D016415:Sequence Alignment / Q000379:methods*; D012689:Sequence Homology, Nucleic Acid; D014170:Transformation, Genetic