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Single-molecule Fluorescence Study for the Effects of DNA Methylation on the Association and Dissociation Kinetics of Restriction Endonuclase and DNA : DNA 메틸화가 제한효소와 DNA 결합 및 분해 동역학에 주는 영향에 대한 단분자 형광 연구
DC Field | Value | Language |
---|---|---|
dc.contributor.advisor | 김성근 | - |
dc.contributor.author | 박소영 | - |
dc.date.accessioned | 2017-07-27T02:18:30Z | - |
dc.date.available | 2017-07-27T02:18:30Z | - |
dc.date.issued | 2014-02 | - |
dc.identifier.other | 000000016655 | - |
dc.identifier.uri | https://hdl.handle.net/10371/134892 | - |
dc.description | 학위논문 (석사)-- 서울대학교 대학원 : 화학부(물리화학전공), 2014. 2. 김성근. | - |
dc.description.abstract | DNA methylation plays a great role both in eukaryotic and prokaryotic cells. In eukaryotes, it suppresses gene regulation, whereas in prokaryotes, specifically in bacteria, it protects the cell from invasion of foreign genes. Many studies have been carried out for the effect of DNA methylation at the ensemble level, but in this study, we investigated the effect of DNA methylation on DNA-protein interaction by single-molecule fluorescence to understand the interaction at the molecular level. We measured the association and dissociation rates of native as well as methylated DNA and found that the values converge to the corresponding ensemble rates. We were able to differentiate the kinetics of association, and dissociation and found that our result was consistent with the fact that DNA methylation interrupts the DNA- protein interaction, especially at a specific kinetic step. | - |
dc.description.tableofcontents | Abstract
1. Introduction 2. Basic principles 2.1 Total internal reflection fluorescence microscopy 2.2 Protein binding induced fluorescence 2.3 Restriction endonuclease, HindIII 2.4 Sample preparation 3. A novel way of detecting protein association and dissociation dynamics and dimer binding of protein. 3.1 Introduction 3.2 Experimental 3.3 Results and discussion 3.3.1 Reaction time in ensemble3.3.2 Enzyme concentration 3.3.3 Specific vs. Nonspecific binding 3.3.4 Single-molecule test 3.4 Conclusion 4. Effects of methylation on dsDNA to dissociation protein from dsDNA. 4.1 Introduction 4.2 Results and discussion 4.2.1 Ensemble test 4.2.2 Single-molecule test 4.2.3 Arrhenius plot 4.2.4 Simulation fitted with single-molecule date 4.3 Conclusion 5. References 6. Appendix 6.1 Ion concentration 6.2 Reaction in imaging buffer 6.3 Enzyme activity on Wild type vs. fluorophore labeled dsDNA 6.4 Time trajectory of a single molecule 국문초록 | - |
dc.format | application/pdf | - |
dc.format.extent | 3872282 bytes | - |
dc.format.medium | application/pdf | - |
dc.language.iso | en | - |
dc.publisher | 서울대학교 대학원 | - |
dc.subject.ddc | 540 | - |
dc.title | Single-molecule Fluorescence Study for the Effects of DNA Methylation on the Association and Dissociation Kinetics of Restriction Endonuclase and DNA | - |
dc.title.alternative | DNA 메틸화가 제한효소와 DNA 결합 및 분해 동역학에 주는 영향에 대한 단분자 형광 연구 | - |
dc.type | Thesis | - |
dc.description.degree | Master | - |
dc.citation.pages | 46 | - |
dc.contributor.affiliation | 자연과학대학 화학부 | - |
dc.date.awarded | 2014-02 | - |
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