Phenol-treatment and a homologous pairing-assay

• Published in: Nucleic acids research Vol. 20; no. 14; pp. 3679 - 3684
• Main Authors: Arai, Naoto, Kawasaki, Katsumi, Iwabuchi, Masa-aki, Shibata, Takehiko
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 25-07-1992
• Subjects: assays; Bacteriophages - genetics; Base Composition - genetics; Biological and medical sciences; Chromatography; DNA; DNA, Circular - metabolism; DNA, Single-Stranded - metabolism; DNA, Superhelical - metabolism; DNA, Viral - metabolism; DNA-Binding Proteins - metabolism; Fundamental and applied biological sciences. Psychology; Fungal Proteins - metabolism; Genic rearrangement. Recombination. Transposable element; Molecular and cellular biology; Molecular genetics; Phenol; Phenols; products; proteins; Rec A Recombinases - metabolism; Recombination, Genetic - genetics; removal; Schizosaccharomyces - metabolism; Schizosaccharomyces pombe; Sequence Homology, Nucleic Acid; treatment; Fungi; Recombination; Yeast; Chromosome pairing; DNA; DNA binding protein; Schizosaccharomyces pombe; Ascomycetes; Thallophyta
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/20.14.3679

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, Schlzosaccharomyces 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 proteinindependent 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 pairingpromoting proteins.