A screening method for DNA aptamers that bind to a specific, unidentified protein in tissue samples.
Aptamers are oligonucleotide ligands with a high affinity to, and specificity for, various target molecules and they are expected to be powerful tools for proteomic analysis. To select aptamers that bind to a specific unidentified protein in tissues for protein analysis, a screening method was developed using chicken skeletal muscle as a model. Target proteins in the target mixture were separated by electrophoresis and transferred to a membrane, and a DNA library was added onto it. The aptamers that bound to the target protein were visualized by chemiluminescence and collected by cutting out the visualized band. The specific aptamers to the target protein were selected by only one round of selection using this screening, suggesting this screening method might be useful for selecting aptamers for proteome analysis.
Noma
Takahisa
T
Department of Life Science and Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16, Naka-cho, Koganei, Tokyo 184-8588, Japan.
Ikebukuro
Kazunori
K
Sode
Koji
K
Ohkubo
Takuya
T
Sakasegawa
Yuji
Y
Hachiya
Naomi
N
Kaneko
Kiyotoshi
K
D000818:Animals; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D002645:Chickens; D015723:Gene Library; D018482:Muscle, Skeletal / Q000378:metabolism; D009841:Oligonucleotides / Q000378:metabolism; D011485:Protein Binding; D040901:Proteomics / Q000379:methods*
D052157:Aptamers, Nucleotide; D009841:Oligonucleotides