| Budget Amount *help |
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000)
Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2012: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Outline of Final Research Achievements |
DNA synthesis and DNA digestion reactions were studied using direct single-molecule observations in microflow channels. In the channels, ssλDNA shape was controlled as relaxed and stretched form by adjusting buffer flow rate, and DNA synthesis reactions by Klenow Fragment (3’-5’ exo-) were observed. Analysis of the synthetic reaction rates demonstrated that the DNA synthesis reaction rate of stretched ssλDNA was approximately 75% higher than that of random coiled ssλDNA . Also, DNA digestion reactions by T7 Exonuclease (T7 Exo) were directly observed both under pulse-chase conditions and under continuous buffer flow conditions with T7 Exo. Under pulse-chase conditions, the double-stranded regions of λDNA monotonously shortened by a DNA digestion reaction with a single T7 Exo molecule. Under continuous buffer flow conditions with T7 Exo, some pauses were observed during a DNA digestion reaction and double-stranded regions shortened linearly except during these pauses.
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