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- Title
Characterization and Application of a Novel RNA Aptamer against the Mouse Prion Protein.
- Authors
Sekiya, Satoru; Noda, Ken; Nishikawa, Fumiko; Yokoyama, Takashi; Kumar, Penmetcha K. R.; Nishikawa, Satoshi
- Abstract
In order to isolate RNA aptamers against the mouse prion protein (mPrP), we carried out in vitro selection from RNA pools containing a 30-nucleotide randomized region. Aptamer 60-3 was found to have a high affinity for mPrP (Kd = 5.6 ± 1.5 nM), and 2′-fluoro-pyrimidine modifications for RNase resistance did not abolish its binding activity (Kd = 22 ± 4 nM). Following 5′ biotinylation, aptamer 60-3 specifically detected PrP in mouse brain homogenate in a Northwestern blotting assay. To determine the mPrP–aptamer binding region, we performed protein-deletion-mutant analysis and competition-binding analysis using heparin. The results showed that aptamer 60-3 appears to have binding sites located between amino acids 23–108.
- Subjects
RNA; NUCLEOTIDES; PYRIMIDINE nucleotides; HEPARIN; AMINO acids
- Publication
Journal of Biochemistry, 2006, Vol 139, Issue 3, p383
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/jb/mvj046