Abstract
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.
| Original language | English |
|---|---|
| Pages (from-to) | 473-480 |
| Number of pages | 8 |
| Journal | Journal of Biomolecular Structure and Dynamics |
| Volume | 25 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - Jan 1 2008 |
| Externally published | Yes |
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All Science Journal Classification (ASJC) codes
- Structural Biology
- Molecular Biology
Cite this
Real-time direct observation of single-molecule dna hydrolysis by exonuclease iii. / Kurita, Hirofumi; Inaishi, Ken Ichi; Torii, Ken; Urisu, Madoka; Nakano, Michihiko; Katsura, Shinji; Mizuno, Akira.
In: Journal of Biomolecular Structure and Dynamics, Vol. 25, No. 5, 01.01.2008, p. 473-480.Research output: Contribution to journal › Article
}
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
PY - 2008/1/1
Y1 - 2008/1/1
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.
UR - http://www.scopus.com/inward/record.url?scp=40949135149&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=40949135149&partnerID=8YFLogxK
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 -