Quantitative Analysis of Werner Helicase Activity Using the Single-Molecule Fluorescence Detection System MF10S(Analytical Biochemistry)
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概要
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We developed a system that uses the single-molecule fluorescence detection system MF10S to assess quantitatively the activity of WRN helicase, the product of the causative gene of Werner syndrome that includes premature ageing. Double-strand DNA substrates labeled with the fluorescence dye TAMRA at the 5' end and with a quencher at the 3' end of the counter strand were incubated with a single trapper oligonucleotide and Werner helicase, and the resultant single DNA fragments labeled with TAMRA produced by the unwinding of WRN helicase were detected using the MF10S. The results using this system and those using polyacrylamide gel electrophoresis were well correlated. The MF10S system provides a quantitative analysis that is much faster, simpler, and more economical than systems using polyacrylamide gel electrophoresis and radioisotopes, and could be used as a quantitative analysis system for Werner helicase and other DNA helicase activities.
- 公益社団法人日本薬学会の論文
- 2005-01-01
著者
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FURUICHI Yasuhiro
GeneCare (previously AGENE) Research Institute
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Furuichi Y
Genecare Research Institute
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Furuichi Yasuhiro
ジーンケア研究所
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Furuichi Yasuhiro
Genecare Research Institute
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FUTAMI Kazunobu
GeneCare Research Institute
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SHIMAMOTO Akira
GeneCare Research Institute
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GOTO Hideyuki
GeneCare Research Institute
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WATANABE Takahide
GeneCare Research Institute Co., Ltd.
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Watanabe Takahide
Department Of Mycology Nippon Roche Research Center
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Watanabe Takahide
Genecare Research Institute Co. Ltd.:core Research For Evolution Science And Technology Japan Scienc
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Furuichi Yasuhiro
Genecare Research Institute Co. Ltd.:core Research For Evolution Science And Technology Japan Scienc
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Watanabe Takahide
Genecare Res. Inst. Co. Ltd.
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