Folding transition of large DNA completely inhibits the action of a restriction endonuclease as revealed by single-chain observation |
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Authors: | Oana Hidehiro Tsumoto Kanta Yoshikawa Yuko Yoshikawa Kenichi |
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Institution: | Department of Microbiology, Philipps-University of Marburg and Max-Planck-Institute for Terrestrial Microbiology, Karl-von-Frisch-Strasse, 35043, Marburg, Germany. |
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Abstract: | The biochemical characteristics of lambda DNA chains in folded/unfolded states upon cleavage by the restriction enzyme ApaLI were investigated in the presence of spermine. These characteristics of DNA chains depending on their higher-order structure were studied at the single-molecule level using fluorescence microscopy. With a low concentration of spermine, lambda DNA takes a random coiled conformation and allows digestion by the enzyme, while under a high concentration of spermine, lambda DNA takes a compact folded structure and inhibits such attack. Together with comparative experiments on short oligomeric DNA, our results suggest that the transition in the higher-order structure causes on/off-type switching of sensitivity to the enzyme. |
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Keywords: | DNA Spermine Restriction enzyme Fluorescence microscopy |
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