A screening method for DNA aptamers that bind to␣a␣specific, unidentified protein in tissue samples

被引:0
作者
Takahisa Noma
Kazunori Ikebukuro
Koji Sode
Takuya Ohkubo
Yuji Sakasegawa
Naomi Hachiya
Kiyotoshi Kaneko
机构
[1] Tokyo University of Agriculture of and Technology,Department of Life Science and Biotechnology
[2] Tokyo Medical University,Second Department Physiology
来源
Biotechnology Letters | 2006年 / 28卷
关键词
Aptamer; Electrophoresis blotting; Multiple target proteins; Systematic evolution of ligands by exponential enrichment;
D O I
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中图分类号
学科分类号
摘要
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.
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页码:1377 / 1381
页数:4
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共 48 条
  • [1] Bianchini M(2001)Specific oligobodies against ERK-2 that recognize both the native and the denatured state of the protein J Immunol Meth 252 191-197
  • [2] Radrizzani M(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
  • [3] Brocardo MG(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
  • [4] Reyes GB(2002)Two-dimensional electrophoresis/phage panning (2D-PP): a novel technology for direct antibody selection on 2-D blots J Biochem 132 245-251
  • [5] Gonzalez Solveyra C(1995)Diversity of oligonucleotide functions Annu Rev Biochem 64 763-797
  • [6] Santa-Coloma TA(2002)Towards proteome-wide production of monoclonal antibody by phage display J Mol Biol 315 1063-1073
  • [7] Blank M(1998)High affinity ligands from in vitro selection: complex targets Proc Natl Acad Sci USA 95 2902-7
  • [8] Weinschenk T(2003)An improved method for the in vitro evolution of aptamers and applications in protein detection and purification Nucleic Acids Res 31 e110-67
  • [9] Priemer M(1986)Regulatory and cytoskeletal proteins of vertebrate skeletal muscle Adv Protein Chem 38 1-370
  • [10] Schluesener H(1997)Nucleic acid selection and the challenge of combinatorial chemistry Chem Rev 97 349-583