Development of biochemical and fine structural analyses methods for higher order chromosome structure

2015 Fiscal Year Final Research Report

• Project/Area Number: 26650010
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Molecular biology
• Research Institution: National Institute of Genetics
• Principal Investigator: ARAKI Hiroyuki 国立遺伝学研究所, 細胞遺伝研究系, 教授 (20151160)
• Co-Investigator(Kenkyū-buntansha): HIZUME Kohji 国立遺伝学研究所, 細胞遺伝研究系, 助教 (10378846)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: クロマチン / DNA複製 / AFM

Outline of Final Research Achievements

The aim of this study is to establish the method of the purification of the chromatin fiber containing specific DNA region and use the chromatin fiber for structural and biochemical analyses. We carried out the fragmentation of chromosomal DNA by the treatment of the purified yeast nuclei with restriction enzymes and the isolation of the specific chromatin fiber by immunoprecipitation. We have also carried out the purification of the plasmid DNA together with the bound proteins from yeast cells. The structure of the chromatin fiber has been visualized and analyzed by atomic force microscopy (AFM). The reconstituted complex consisting of the chromatin fiber and DNA-replication factors has also been visualized by AFM to evaluate the size and shape of the particles observed in the chromatin fiber purified from yeast cells.

Free Research Field

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