Phenol-treatment and a homologous pairing-assay.
Homologous pairing is a key step in homologous genetic recombination. In the early stage of trials for the identification of homologous pairing-promoting proteins from a fission yeast, Schizosaccharomyces pombe, we treated DNA products with phenol in the presence of a salt for the removal of tightly bound proteins from DNA before the assay, but we found that this treatment caused very efficient protein-independent double-strand formation from complementary single-stranded DNAs. Using an assay including the phenol treatment, we detected another species of apparent homologous pairing-promoting proteins in the nuclei, in addition to a homologous pairing-promoting protein consisting of three components which we reported previously. However, studies involving the use of an assay without the phenol-treatments revealed that the second one was not really a homologous pairing-protein. Thus, the protein-independent double-strand formation by phenol-treatment in the presence of a salt could cause the erroneous identification of homologous pairing-promoting proteins.
Arai
N
N
Laboratory of Microbiology, Institute of Physical and Chemical Research (RIKEN), Saitama, Japan.
Kawasaki
K
K
Iwabuchi
M
M
Shibata
T
T
D001435:Bacteriophages / Q000235:genetics; D001482:Base Composition / Q000235:genetics; D002845:Chromatography; D004270:DNA, Circular / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism*; D004278:DNA, Superhelical / Q000378:metabolism; D004279:DNA, Viral / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005656:Fungal Proteins / Q000378:metabolism*; D019800:Phenol; D010636:Phenols; D011938:Rec A Recombinases / Q000378:metabolism*; D011995:Recombination, Genetic / Q000235:genetics; D012568:Schizosaccharomyces / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D004270:DNA, Circular; D004277:DNA, Single-Stranded; D004278:DNA, Superhelical; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D010636:Phenols; D019800:Phenol; D011938:Rec A Recombinases