Evaluation of small ligand-protein interaction by ligation reaction with DNA-modified ligand.
A method for the evaluation of interactions between protein and ligand using DNA-modified ligands, including signal enhancement of the DNA ligation reactions, is described. For proof of principle, a DNA probe modified by biotin was used. Two DNA probes were prepared with complementary sticky-ends. While one DNA probe was modified at the 5'-end of the sticky-end, the other was not modified. The probes could be ligated together by T4 DNA ligase along the strand without biotin modification. However, in the presence of streptavidin or anti-biotin Fab, the ligation reaction joining the two probes could not occur on either strand.
Sugita
Rie
R
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama, 226-8501, Japan.
Mie
Masayasu
M
Funabashi
Hisakage
H
Kobatake
Eiry
E
D001710:Biotin / Q000737:chemistry; D004247:DNA / Q000737:chemistry* / Q000378:metabolism*; D011088:DNA Ligases / Q000378:metabolism; D015342:DNA Probes / Q000737:chemistry / Q000378:metabolism; D004076:Digoxigenin / Q000737:chemistry; D008954:Models, Biological; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D011506:Proteins / Q000378:metabolism*
D015342:DNA Probes; D011506:Proteins; D001710:Biotin; D004247:DNA; D011088:DNA Ligases; D004076:Digoxigenin