| Project/Area Number |
14340238
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
遺伝
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
SHIMIZU noriaki Hiroshima University, Faculty of Integrated Arts and Sciences, Associate Professor, 総合科学部, 助教授 (10216096)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥10,500,000 (Direct Cost: ¥10,500,000)
Fiscal Year 2004: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2003: ¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥3,700,000 (Direct Cost: ¥3,700,000)
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| Keywords | Gene Amplification / Extrachromosomal Element / Replication Initiation Region / Matrix Attachment Region / Micronuclei / DMs / HSR / 生細胞可視化技術 / 2本鎖切断 / DNA修復 / 細胞周期チェックポイント / 細胞内動態 / BFB cycle / anaphase bridge / 複製 / 哺乳動物複製起点 / ポリ(A)付加配列 / 複製フォーク阻害配列 |
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| Research Abstract |
Gene amplification plays a pivotal role in malignant transformation of human cells. The amplified genes localize either at extrachromsomal DMsorchromosomal HSR. DMs are autonomously replicating acentric extrachromosomal chromatin. We previously found that the plasmid having a mammalian replication initiation region and a matrix attachment region efficiently generated DMs and/or HSR, de novo. We also found that the human cancer cells lose tumor phenotype and normalized, if the DMs were eliminated. The elimination was mediated by the specific incorporation of DMs into the cytoplasmic micronuclei. The inclusion into the micronuclei depended on a novel intracellular behavior of DMs during the cell cycle progression. The behavior and the elimination of DMs is common among abroad spectrum of autonomously replicating extrachromosomal genetic element. In this study, we advanced our understanding upon the molecular mechanism that govern the behavior and the elimination of extrachromosomal element. Furthermore, we applied the understanding to the basic biological study on the nucleus, as well as to the technology to produce recombinant protein. Specifically, we did the following studies, and obtained many achievements from each of these. 1) Study on the molecular mechanism underlying the intracellular behavior of DMs that led to the inclusion into the micronuclei. 2) Study on the behavior and the elimination of DNA molecule placed at the nucleus or the cytoplasm by microinjection. 3) Study on the mechanism of gene amplification mediated by the autonomously replicating extrachromosomal molecule. 4) Study on the gene expression from the amplified genetic region. 5) Study on the application of our efficient gene amplification system to the technology on recombinant protein production. 6) Study on the functional structure of the interphase cell nucleus by using a novel feature of HSR that was generated by our plasmid-based amplification system.
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