Efficient interaction of recA protein with fluorescent dye‐labeled oligonucleotides

FEBS Letters

Bocharova, Tatjana N.¹  Volodin, Alexander A.¹  Smirnova, Helen A.¹ 

[1] Institute of Molecular Genetics of the Russian Academy of Sciences, Kurchatov Sq., Moscow, 123182, Russian Federation

关键词: recA protein;    Fluorescent dye-labeled oligonucleotide;    recA-oligonucleotide complex;    Strand exchange;    ATP-S;    adenosine-5′-O-(3-thiotriphosphate);    JOE;    4′;    5′-dichloro-2′;    7′-dimethoxy-6-carboxy-fluorescein;    PAGE;    polyacryl-amide gel electrophoresis;    ds-;    ss-;    double- and single-stranded unmodified oligonucleotides respectively;    ds*-;    ss*-;    double- and single stranded dye-labeled oligonucleotides respectively;    b.;    base;    bp;    base pair;    SDS;    dodecyl sulfate sodium salt;   

DOI  :  10.1016/0014-5793(94)00640-7

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Some fluorescein derivatives attached to the 5′-end of oligonucleotides stimulate recA protein—oligonucleotide binding. The complex formation at near stoichiometric DNA/protein ratios is demonstrated for 18-bases-long oligonucleotides. The complexes with dye-labeled oligonucleotides are shown to be active in the reaction of homologous strand exchange. The strand exchange reaction in the presence of adenosine-5′-O-(3-thiotriphosphate) proceeds with the formation of a stable complex of recA protein with the double stranded oligonucleotide, which is a product of the strand exchange. The displaced single-stranded oligonucleotide is shown to be bound weakly.

【 授权许可】

Unknown