Efficient interaction of recA protein with fluorescent dye-labeled oligonucleotides.
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.
Volodin
A A
AA
Institute of Molecular Genetics, Russian Academy of Sciences, Moscow.
Smirnova
H A
HA
Bocharova
T N
TN
D001483:Base Sequence; D004277:DNA, Single-Stranded / Q000378:metabolism*; D005452:Fluoresceins / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism*; D011938:Rec A Recombinases / Q000378:metabolism*; D011995:Recombination, Genetic
D004277:DNA, Single-Stranded; D005452:Fluoresceins; D005456:Fluorescent Dyes; D009838:Oligodeoxyribonucleotides; D011938:Rec A Recombinases