Abstract
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.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Bianchini M, Radrizzani M, Brocardo MG, Reyes GB, Gonzalez Solveyra C, Santa-Coloma TA (2001) Specific oligobodies against ERK-2 that recognize both the native and the denatured state of the protein. J Immunol Meth 252:191–197
Blank M, Weinschenk T, Priemer M, Schluesener H (2001) Systematic evolution of a DNA aptamer binding to rat brain tumor microvessels. Selective targeting of endothelial regulatory protein pigpen. J Biol Chem 276:16464–16468
Daniels DA, Chen H, Hicke BJ, Swiderek KM, Gold L (2003) A tenascin-C aptamer identified by tumor cell SELEX: systematic evolution of ligands by exponential enrichment. Proc Natl Acad Sci USA 100:15416–15421
Furuta M, Ito T, Eguchi C, Tanaka T, Wakabayashi-Takai E, Kaneko K (2002) Two-dimensional electrophoresis/phage panning (2D-PP): a novel technology for direct antibody selection on 2-D blots. J Biochem 132:245–251
Gold L, Polisky B, Uhlenbeck O, Yarus M (1995) Diversity of oligonucleotide functions. Annu Rev Biochem 64:763–797
Liu B, Huang L, Sihlbom C, Burlingame A, Marks JD (2002) Towards proteome-wide production of monoclonal antibody by phage display. J Mol Biol 315:1063–1073
Morris KN, Jensen KB, Julin CM, Weil M, Gold L. (1998) High affinity ligands from in vitro selection: complex targets. Proc Natl Acad Sci USA 95:2902–7
Murphy MB, Fuller ST, Richardson PM, Doyle SA (2003) An improved method for the in vitro evolution of aptamers and applications in protein detection and purification. Nucleic Acids Res 31:e110
Ohtsuki I, Maruyama K, Ebashi S (1986) Regulatory and cytoskeletal proteins of vertebrate skeletal muscle. Adv Protein Chem 38:1–67
Osborne SE, Ellington AD (1997) Nucleic acid selection and the challenge of combinatorial chemistry. Chem Rev 97:349–370
Wang J, Jiang H, Liu F (2000) In vitro selection of novel RNA ligands that bind human cytomegalovirus and block viral infection. RNA 6:571–583
Zuker M (2003) Mfold web server for nucleic acid folding and hybridization prediction. Nucleic Acids Res. 31:3406–3415
Acknowledgement
This work is partly supported by the Grant-in-Aid for the 21st Century COE “Future Nano-Materials” from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Noma, T., Ikebukuro, K., Sode, K. et al. A screening method for DNA aptamers that bind to␣a␣specific, unidentified protein in tissue samples. Biotechnol Lett 28, 1377–1381 (2006). https://doi.org/10.1007/s10529-006-9106-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-006-9106-7