TY - JOUR
T1 - Real-time direct observation of single-molecule dna hydrolysis by exonuclease iii
AU - Kurita, Hirofumi
AU - Inaishi, Ken Ichi
AU - Torii, Ken
AU - Urisu, Madoka
AU - Nakano, Michihiko
AU - Katsura, Shinji
AU - Mizuno, Akira
N1 - Funding Information:
This work was supported in part by Grants-in-Aid for Scientific Research of Priority Areas “System Cell Engineering by Multi-scale Manipulation” from the Ministry of Education, Culture, Sports, Science and Technology (MEXT) of Japan, the Japan Society for the Promotion of Science (JSPS) Research Fellowships for Young Scientists (18·50512), and the MEXT 21st Century COE Program “Ecological Engineering for Homeostatic Human Activities.”
PY - 2008/4
Y1 - 2008/4
N2 - Single-molecule DNA digestion by exonuclease III, which has 3′ to 5′ exonuclease activity, was analyzed using a micro-channel with two-layer laminar flow. First, a DNA-bead complex was optically trapped in one layer in the absence of exonuclease III permitted the DNA to be stretched by the laminar flow. The exonuclease III reaction was initiated by moving the trapped DNA-bead complex to another layer of flow, which contained exonuclease III. As the reaction proceeded, the fluorescently-stained DNA was observed to shorten. The process was photographed; examination of the photographs showed that the DNA molecule shortened in a linear fashion with respect to the reaction time. The digestion rate obtained from the single-molecule experiment was compared to that measured from a bulk experiment and was found to be ca. 28 times higher than the bulk digestion rate.
AB - Single-molecule DNA digestion by exonuclease III, which has 3′ to 5′ exonuclease activity, was analyzed using a micro-channel with two-layer laminar flow. First, a DNA-bead complex was optically trapped in one layer in the absence of exonuclease III permitted the DNA to be stretched by the laminar flow. The exonuclease III reaction was initiated by moving the trapped DNA-bead complex to another layer of flow, which contained exonuclease III. As the reaction proceeded, the fluorescently-stained DNA was observed to shorten. The process was photographed; examination of the photographs showed that the DNA molecule shortened in a linear fashion with respect to the reaction time. The digestion rate obtained from the single-molecule experiment was compared to that measured from a bulk experiment and was found to be ca. 28 times higher than the bulk digestion rate.
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U2 - 10.1080/07391102.2008.10507194
DO - 10.1080/07391102.2008.10507194
M3 - Article
C2 - 18282002
AN - SCOPUS:40949135149
VL - 25
SP - 473
EP - 480
JO - Journal of Biomolecular Structure and Dynamics
JF - Journal of Biomolecular Structure and Dynamics
SN - 0739-1102
IS - 5
ER -