Construction and characterization of a rad51rad52 double mutant as a host for YAC libraries.
RAD52 or RAD51 recombination-deficient yeast strains stabilize otherwise unstable YACs containing ribosomal DNA or the human color vision locus (Kohno et al., 1994). Thus the RAD52RAD51 pathways(s) are apparently involved in the instability of YACs containing tandem repeat loci, presumably by promoting recombination-based deletion formation. Some other genomic loci are still unstable or unrecoverable in those strains, but we now find that greater stability is observed in a rad51rad52 double mutant strain that we have newly constructed. YACs containing a highly unstable region around DXS49 or centromeric regions throw off a variety of products in single mutants, but are much more stable in the rad51rad52 strain, which could therefore provide a better host for library construction and maintenance.
Kohno
K
K
Department of Molecular and Cellular Biology for Pharmaceutical Sciences, Kyoto Pharmaceutical University, Misasagi, Yamashina-ku, Japan. kykohno@mb.kyoto-phu.ac.jp
Oshiro
T
T
Kishine
H
H
Wada
M
M
Takeda
H
H
Ihara
N
N
Imamoto
F
F
Kano
Y
Y
Schlessinger
D
D
D018244:Chromosomes, Artificial, Yeast; D004268:DNA-Binding Proteins / Q000235:genetics*; D005656:Fungal Proteins / Q000235:genetics*; D015723:Gene Library; D006801:Humans; D009154:Mutation; D010957:Plasmids; D051135:Rad51 Recombinase; D051721:Rad52 DNA Repair and Recombination Protein; D012441:Saccharomyces cerevisiae / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins; D013045:Species Specificity
D004268:DNA-Binding Proteins; D005656:Fungal Proteins; C497197:RAD52 protein, S cerevisiae; D051721:Rad52 DNA Repair and Recombination Protein; D029701:Saccharomyces cerevisiae Proteins; C495151:RAD51 protein, S cerevisiae; C495153:RAD51 protein, human; D051135:Rad51 Recombinase