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Structural analysis of nucleic acids by labeled oligonucleotides.
A Murakami1, S Nagahara, M Nakaura
1Department of Polymer Science and Engineering, Faculty of Textile Science, Kyoto Institute of Technology, Japan.
Nucleic Acids Symposium Series
|January 1, 1990
Summary
Labeled oligonucleotides (S-probes and F-probes) were characterized for nucleic acid structure analysis. Their mobility and structural changes indicate they can recognize and analyze nucleic acid secondary structures in solution.
Area of Science:
- Biochemistry
- Molecular Biology
- Biophysical Chemistry
Background:
- Characterizing nucleic acid structure in solution is crucial for understanding biological function.
- Labeled oligonucleotides offer potential tools for such analyses.
Purpose of the Study:
- To characterize spin-labeled and fluorescent-labeled oligonucleotides for nucleic acid structural analysis.
- To assess the utility of these probes in recognizing secondary structures of nucleic acids in solution.
Main Methods:
- Site-specific spin labeling of oligonucleotides with TEMPO (S-probe).
- Fluorescent labeling of oligonucleotides with fluorescein (F-probe).
- Electron Spin Resonance (ESR) and fluorescence depolarization analysis.
Main Results:
- ESR line changes and rotational correlation time (tau) of S-probes were sensitive to base sequence, hybridization, and conformation.
- Tau values reflected the local structure of oligonucleotide hybrids.
- S-probes and F-probes demonstrated recognition of 5S RNA secondary structure in solution.
Conclusions:
- Labeled oligonucleotides (S-probes and F-probes) are effective for analyzing nucleic acid secondary structures in solution.
- These probes can be used to study the conformation and hybridization of nucleic acids.