Selection of a DNA aptamer that binds 8-OHdG using GMP-agarose.
DNA aptamers, which bind specific molecule, such as 8-OHdG, with high affinity were investigated using an in vitro selection strategy called systematic evolution of ligands by exponential enrichment (SELEX). However, 8-OHdG was difficult to immobilize on a carrier for SELEX. Therefore, a DNA aptamer binding to 8-OHdG was selected using GMP-agarose as an analogue from a library of about 4(60) random ssDNA sources. As a result, three aptamer candidates were selected. Among the selected DNA aptamers, the No. 22 DNA aptamer exhibited a high affinity for 8-OHdG. The dissociation constant, K(D), of No. 22 DNA aptamer was on the order of 0.1micromol/L. This result suggests that using an analogue will be a useful new SELEX method for obtaining various aptamers that are difficult to immobilize on a matrix.
Miyachi
Yusuke
Y
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, Rokkoudai-chou 1-1, Nada, Kobe 657-8501, Japan.
Shimizu
Nobuaki
N
Ogino
Chiaki
C
Fukuda
Hideki
H
Kondo
Akihiko
A
D000080242:8-Hydroxy-2'-Deoxyguanosine; D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000302:isolation & purification; D001483:Base Sequence; D002942:Circular Dichroism; D004277:DNA, Single-Stranded / Q000737:chemistry; D003849:Deoxyguanosine / Q000031:analogs & derivatives* / Q000737:chemistry; D006157:Guanosine Monophosphate / Q000737:chemistry*; D007700:Kinetics; D009690:Nucleic Acid Conformation; D052156:SELEX Aptamer Technique / Q000379:methods*; D012685:Sepharose
D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D006157:Guanosine Monophosphate; D000080242:8-Hydroxy-2'-Deoxyguanosine; D012685:Sepharose; D003849:Deoxyguanosine