An aptamer-based biosensor for mammalian initiation factor eukaryotic initiation factor 4A.
Aptamers are short single-stranded DNA or RNA sequences that are selected in vitro based on their high affinity to a target molecule. Here we demonstrate that an RNA aptamer selected against eukaryotic initiation factor 4A (eIF4A) serves as an efficient biosensor. The aptamer, when immobilized to resin, purifies eIF4A from crude cell extracts by affinity pull-down, and 32P-labeled aptamer can detect some 300 ng of eIF4A by dot-blot analysis. Moreover, by use of an aptamer-immobilized sensor chip, we developed a surface plasmon resonance assay to detect eIF4A at the nanogram level within whole cell lysates after optimizing sample preparation, thereby showing a real-time sensor for eIF4A in cell extract solution.
Oguro
Akihiro
A
Department of Basic Medical Sciences, Institute of Medical Science, University of Tokyo, Minato-ku, Tokyo 108-8639, Japan.
Ohtsu
Takashi
T
Nakamura
Yoshikazu
Y
D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D004277:DNA, Single-Stranded / Q000737:chemistry; D039601:Eukaryotic Initiation Factor-4A / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D015151:Immunoblotting
D052157:Aptamers, Nucleotide; D004277:DNA, Single-Stranded; D039601:Eukaryotic Initiation Factor-4A