Application of microchip electrophoresis in the analysis of RNA aptamer-protein interactions.
DNA and RNA can be separated by microchip electrophoresis (ME) and detected using an intercalating fluorescent dye. The advantages of this method are short sensing times (<3 min), avoidance of a radioisotope labeling detection system, relatively low costs, and reduced labor intensity. In the present study, RNA aptamer-protein or -peptide interactions were analyzed using ME and the regression of free aptamers corresponding to unbound RNA was detected as the target protein or peptide increased in a dose-dependent manner. Our results demonstrate the applicability of this method to simple, rapid ligand screening in the interactions between oligonucleotides and their targets.
Nishikawa
Fumiko
F
Institute for Biological Resources and Functions, National Institute of Advanced Industrial Science and Technology, Tsukuba, Ibaraki, Japan. satoshi-nishikawa@aist.go.jp
Arakawa
Hidetoshi
H
Nishikawa
Satoshi
S
D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000378:metabolism*; D001483:Base Sequence; D019075:Electrophoresis, Capillary; D046209:Electrophoresis, Microchip / Q000379:methods*; D007700:Kinetics; D008024:Ligands; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D012313:RNA / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
D052157:Aptamers, Nucleotide; D008024:Ligands; D011506:Proteins; D012313:RNA