1995 Fiscal Year Final Research Report Summary
Chemical Creation of Artificial Restriction Enzyme and Its Application to Genomic Analysis
| Project/Area Number |
05557111
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
医薬分子機能学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
SUGIURA Yukio Kyoto Univ.Professor, 化学研究所, 教授 (40025698)
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| Co-Investigator(Kenkyū-buntansha) |
OTSUKA Masami Kyoto Univ.Associate Professor, 化学研究所, 助教授 (40126008)
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| Project Period (FY) |
1993 – 1995
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| Keywords | Artificial Restriction Enzymo / Bleomycin / DNA Cutting / Zinc Finger / Nuclease / Triple Stranded DNA / Human Geome |
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| Research Abstract |
The molecular mechanism of bleomycin antibiotics facilitated design and synthesis of srtificial DNA cleaving molecules. Transformation of a sequence-specific DNA-binding molecule into a sequence-specific DNA-cleaving molecule allows identification of the preferred binding locations of these molecules on restriction fragments analyzed on sequencing gels. Various types of DNA-cleaving molecules have been demonstrated. Sequence-specific DNA-cleaving peptides and oligonucleotides equipped with a DNA-cleaving moiety have also been designed. Zinc finge motif is a novel DNA binding motif characterized by the unique role of zinc ; the protein folding and the DNA binding ability are goverened by the coordination of a zinc ion. In C_2H_2-type zinc finger, two cysteines and two histidines contribute to form a globular domain through zinc coordination. The zinc finger of C_2H_2-type gives promise of recognition for any DNA sequences because of its recognition mode. A C_2H_2-type zinc finger protein Sp1 has been converted into artificial sitespecific nuclease with an attached Ni-based DNA cleavage unit (Gly-Gly-His). This protein cleaved DNA at a single site near the Sp1 recognition sequence. The zinc finger-based nuclease is applicable to chromosome mapping and sequencing.
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