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Identification of Replication Units in Mammalian Genome by Fiber-FISH

Research Project

Project/Area Number 09660080
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionMie University

Principal Investigator

OKUMURA Katsuzumi  Mie Univ.Bioresources Associate Prof., 生物資源学部, 助教授 (30177183)

Project Period (FY) 1997 – 1998
Project Status Completed (Fiscal Year 1998)
Budget Amount *help
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 1998: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 1997: ¥2,200,000 (Direct Cost: ¥2,200,000)
KeywordsDNA replication / FISH / DNA fiber / Replicon / Chromatin structure / Bromodeoxyuridine / PWS / AS / replication timing / intranuclear structures
Research Abstract

Florescence in situ hybridization (FISH) is a powerful tool for gene mapping as well as biological analyses. In this work, the FISH technique was applied to the analysis of DNA replication process in the animal cell genome. Fiber-FISH allows us to analyze the genomic distances in several kb level. Newly synthesized DNAs can be chased on a DNA fiber by detecting incorporated bromodeoxyuridine (BrdU), a thymidine analog. The primary purpose in this study is to examine the combined method of these two fine techniques and its feasibility for the analysis of DNA replication process. Firstly, the detection method of the incorporated BrdU was improved. Alkaline danaturation of DNA was found to detect the replicating DNA fiber uniformly and efficiently. The progress of the DNA replication on a DNA fiber was chased in a time dependent manner. Secondly, in this condition, the genomic DNA could be visualized by FISH.The simultaneous detection of newly synthesized DNA and specific genome sequences was also possible. A highly synchronizing method of HL60 cells was also developed. These results indicate that replicons and replicon cluster domains can be determined by the application of these finetechniques. As a related studies, a method for analysis of a packaging status of the specific genome sequences was developed. This is an important for the understanding of the intranuclear genome organization including DNA replication domains. Intranuclear packaging status of the genomic clones could be visualized on a nuclear halo. This technique was applied to the imprinted gene, SNRPN, suggesting that a SNRPN gene on the paternal allele is more compacted than that on the maternal allele. These information is very important for clarifying the DNA replication mechanismin relation to the gene expression and chromatin structures.

Report

(3 results)
  • 1998 Annual Research Report   Final Research Report Summary
  • 1997 Annual Research Report
  • Research Products

    (63 results)

All Other

All Publications (63 results)

  • [Publications] Takayuki Hattori: "Molecular Cloning of a cDNA Encoding Human TSC-22 (Transforming Growth Factor b1 Stimulated Clone 22) and Localization of the Gene at Chromosome13q14" Acta Sholae Medicinalis Universitatis in Gifu. 45. 201-208 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nobuhisa Mizuki: "Nucleotide Sequence Analysis of the HLA Class I Region Spanning the 237 kb Segment Around the HLA-B and -C Genes" Genomics. 42. 55-66 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Toyoaki Tenzen: "Precise Switching of DNA Replication Timing in the GC Content Transition Area in the Human Major Histocompatibility Complex" Molecular and Cellular Biology. 17. 4043-4050 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masamichi Ishiai: "Isolation of Human and Fission Yeast Gene Homologues of the Budding Yeast Origin Recognition Complex Subunit Orc5 : Human Homologue (ORC5L) Maps to 7q22" Genomics. 46. 294-298 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Toshihiko Eki: "Mapping of the Human Genes Encoding Cyclin H and the CDK-activating Kinase Assembly Factor MAT1 (MNAT1) to Chromosome Bands 5q13.3-q14 and 14q23, Respectively :" Genomics. 47. 115-120 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masashi Kimura: "Assignment of STK6 to Human Chromosome 20q13.2-13.3 and a Pseudogene STK6P to 1q41-42" Cytogenetics and Cell Genetics. 79. 201-203 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] 奥村克純: "FISHによる動物細胞ゲノムの核内構造解析の現状" 生化学. 69. 41-46 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Chee-Gun Lee: "Molecular Analysis of the cDNA and Genomic DNA Encoding Mouse RNA Helicase A" Genomics. 47. 365-371 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sachiko Kitanaka: "Inactivating Mutations in the 25-Hydroxyvitamin D3 1α-Hydroxylase gene in Patients with Pseudovitamin D-Deficiency Rickets" New England Journal of Medicine. 338. 653-661 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Katsuzumi Okumura: "Temporal Order of DNA Replication of Imprinted IGF2 Gene and Non-imprinted TNF and RFC5 Genes in Human Lymphocytes" Bulletin of the Faculty of Bioresources Mie University. 20号. 47-54 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yasushi Taniguchi: "Nucleotide Sequence of the Ring3 Gene in the Class II Region of the Mouse MHC and Its Abundant Expression in Testicular Germ Cells" Genomics. 51. 114-123 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Toshihiko Eki: "Assignment of the Human Homologue of the Yeast Origin Recognition Complex Subunit ORC4 to Human Chromosome Band 2q22-q23 by in situ Hybridization and Somatic Cell Hybrid Analysis" Cytogenetics and Cell Genetics. 81. 89-90 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Noriyuki Yanaka: "Expression, Structure and Chromosomal Localization of the Human cGMP-binding cGMP-specific Phosphodiesterase Gene(PDE5A)" European Journal of Biochemistry. 25. 391-399 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takashi Shiina: "Physical Mapping between the S and HLA-E Genes in the Human MHC Class I Region : Construction of a BAC, PAC and Cosmid Contig" Immunogenetics. 48. 402-407 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Katsuzumi Okumura: "Chromosomal Localization of Human DNA-binding Nuclear Protein (NP220) to 2p13.1-p13.2 by Fluorescence in situ Hybridization and Its Relation to Matrin 3" Bioscience, Biotechnology and Biochemistry. 62. 1640-1642 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shin Ogata: "Apoptosis Induced by Niacin-related Compounds in HL-60 Cells" Bioscience, Biotechnology and Biochemistry. 62. 2351-2356 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takeshi Uchiumi: "Isolation of a Novel Human Canalicular Multispecific Organic Anion Transporter, cMOAT2/MRP3 and Its Expression in Cisplatin Resistant Cancer Cells with Decreased ATP-dependent Drug Transport" Biochemical Biophysical Research Commnications. 252. 103-110 (1998)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shin Ogata: "Effects of Poly(ADP-ribose)polymerase Inhibitors and NAD on the Apoptosis Induced by Actinomycin D in Human Leukemia Cells HL-60" Bulletin of the Faculty of Bioresources Mie University. 21号. 73-85 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masato Tsurudome: "Primary Structure of the Light Chain of FRP-1/CD98/4F2 Predicts a Protein with 11 Transmembrane Domains" Journal of Immunology. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kotomi Fujishige: "Cloning and characterization of a novel human cAMP and cAMP-inhibited cGMP phosphodiesterase (PDE10A)" Journal of Biological Chemistry. (印刷中).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takayuki Hattori, et al.: "Molecular Cloning of a cDNA Encoding Human TSC-22 (Transforming Growth Factor beta1-Stimulated Clone 22) and Localization of the Gene at Chromosome 13q14" Acta Sholae Medicinalis Universitatis in Gifu. 45 (2). 201-208 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Nobuhisa Mizuki, et al.: "Nucleotide Sequence Analysis of the HLA Class I Region Spanning the 237kb Segment Around the HLA-B and-C Genes" Genomics. 42 (1). 55-66 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Toyoaki Tenzen, et al.: "Precise Switching of DNA Replication Timing in the GC Content Transition Area in the Human Major Hisocompatibility Complex" Mol.Cell.Biol.17 (7). 4043-4050 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masamichi Ishiai, et al.: "Isolation of Human and Fission Yeast Gene Homologues of the Budding Yeast Origin Recognition Complex Subunit Orc5 : Human Homologue (ORC5L) Maps to 7q22" Genomics. 46. 294-298 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Toshihiko Eki, et al.: "Mapping of the Human Genes Encoding Cyclin H (CCNH) and the CDK-activating Kinase (CAK) Assembly Factor MAT1 (MNAT1) to Chromosome Bands 5q13.3-q14 and 14q23, Respectively" Genomics. 47. 115-120 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masashi Kimura, et al.: "Assignment of STK6 to Human Chromosome 20q13.2-13.3 and a Pseudogene STK6P to 1q41-42" Cytogenet.Cell Genet.79. 201-203 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Chee-Gun Lee, et al.: "Molecular Analysis of the cDNA and Genomic DNA Encoding Mouse RNA Helicase A" Genomics. 47 (1). 365-371 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Sachiko Kitanaka, et al.: "Inactivating Mutations in the 25-Hydroxyvitamin D_3 1alpha-Hydroxylase gene in Patients with Pseudovitamin D-Deficiency Rickets" New England J.of Med.338 (10). 653-661 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Katsuzumi Okumura, et al.: "Temporal Order of DNA Replication of Imprineted IGF2 Gene and Non-imprinted TNF and RFC5 Genes in Human Lymphocytes" Bull.Fac.Biores.Mie Univ.20. 47-54 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Yasushi Taniguchi, et al.: "Nucleotide Sequence of the Ring3 Gene in the Class II Region of the Mouse MHC and Its Abundant Expression in Testicular Germ Cells" Genomics. 51. 114-123 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Toshihiko Eki, et al.: "Assignment of the Human Homologue of the Yeast Origin Recognition Complex Subunit ORC4 (ORC4L) to Human Chromosome Band 2q22-q23 by in situ Hybridization and Somatic Cell Hybrid Analysis" Cytogenet.Cell Genet.81. 89-90 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Noriyuki Yanaka, et al.: "Expression, Structure and Chromosomal Localization of the Human cGMP-binding cGMP-specific Phosphodiesterase Gene (PDE5A)" Eur.J.Biochem.25 (2). 391-399 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takashi Shiina, et al.: "Physical Mapping between the S and HLA-E Genes in the Human MHC Class I Region : Construction of a BAC,PAC and Cosmid Contig" Immunogenetics. 48. 402-407 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Katsuzumi Okumura, et al.: "Chromosomal Localization of Human DNA-binding Nuclear Protein (NP220) to 2p13.1-p13.2 by Fluorescence in situ Hybridization and Its Relation to Matrin 3" Biosci.Biotech.Biochem.62 (8). 1640-1642 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shin Ogata, et al.: "Apoptosis Induced by Niacin-related Compounds in HL-60 Cells" Biosci.Biotech.Biochem.62 (12). 2351-2356 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Takeshi Uchiumi, et al.: "Isolation of a Novel Human Canalicular Multispecific Organic Anion Transporter, cMOAT2/MRP3 and Its Expression in Cisplatin Resistant Cancer Cells with Decreased ATP-dependent Drug Transport" Biochem.Biophys.Res.Commn.252. 103-110 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Shin Ogata, et al.: "Effects of Poly (ADP-ribose) polymerase Inhibitors and NAD on the Apoptosis Induced by Actinomycin D in Human Leukemia Cells HL-60" Bull.Fac.Biores.Mie Univ.21. 73-85 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Masato Tsurudome, et al.: "Primary Structure of the Light Chain of FRP-1/CD98/4F2 Predicts a Protein with 11 Transmembrane Domains" J.Immunol.(in press.).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Kotomi Fujishige, et al.: "Cloning and characterization of a novel human cAMP and cAMP-inhibited cGMP phosphodiesterase (PDE10A)" J.Biol.Chem.(in press.).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Katsuzumi Okumura: "Recent Progress in the Analysis of the Intranuclear Organization of the Animal Cell Genome by FISH" Seikagaku. 69 (5). 41-46 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1998 Final Research Report Summary
  • [Publications] Chee-Gun Lee: "Molecular Analysis of the cDNA and Genomic DNA Encoding Mouse RNA Helicase A" Genomics. 47巻. 365-371 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Sachiko Kitanaka: "Inactivating Mutations in the 25-Hydroxyvitamin D3 1α-Hydroxylase gene in Patients with Pseudovitamin D-Deficiency Rickets" New England Journal of Medicine. 338巻. 653-661 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Katsuzumi Okumura: "Temporal Order of DNA Replication of Imprinted IGF2 Gene and Non-imprinted TNF and RFC5 Genes in Human Lymphocytes" Bulletin of the Faculty of Bioresources Mie University. 20号. 47-54 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Yasushi Taniguchi: "Nucleotide Sequence of the Ring3 Gene in the Class II Region of the Mouse MHC and Its Abundant Expression in Testicular Germ Cells" Genomics. 51巻. 114-123 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Toshihiko Eki: "Assignment of the Human Homologue of the Yeast Origin Recognition Complex Subunit ORC4 to Human Chromosome Band 2q22-q23 by in situ Hybridization and Somatic Cell Hybrid Analysis" Cytogenetics and Cell Genetics. 81巻. 89-90 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Noriyuki Yanaka: "Expression,Structure and Chromosomal Localization of the Human cGMP-binding cGMP-specific Phosphodiesterase Gene(PDE5A)" European Journal of Biochemistry. 25巻. 391-399 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takashi Shiina: "Physical Mapping between the S and HLA-E Genes in the Human MHC Class I Region:Construction of a BAC,PAC and Cosmid Contig" Immunogenetics. 48巻. 402-407 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Katsuzumi Okumura: "Chromosomal Localization of Human DNA-binding Nuclear Protein(NP220)to 2P13.1-p13.2 by Fluorescence in situ Hybridization" Bioscience,Biotechnology and Biochemistry. 62巻. 1640-1642 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shin Ogata: "Apoptosis Induced by Niacin-related Compounds in HL-60 Cells" Bioscience,Biotechnology and Biochemistry. 62巻. 2351-2356 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Takeshi Uchiumi: "Isolation of Novel Human Canalicular Multispecific Organic Anion Transporter,cMOAT2/MRP3 and Its Expression in Cisplatin Resistant Cancer Cells with Decreased ATP-dependent Drug Transport" Biochemical Biophysical Research Communications. 252巻. 103-110 (1998)

    • Related Report
      1998 Annual Research Report
  • [Publications] Shin Ogata: "Effects of Poly(ADP-ribose)polymerase Inhibitors and NAD on the Apoptosis Induced by Actinomycin D in Human Leukemia Cells HL-60" Bulletin of the Faculty of Bioresources Mie University. (印刷中).

    • Related Report
      1998 Annual Research Report
  • [Publications] Masato Tsurudome: "Primary Structure of the Light Chain of FRP-1/CD98/4F2 Predicts a Protein with 11 Transmembrane Domains" Journal of Immunology. (印刷中).

    • Related Report
      1998 Annual Research Report
  • [Publications] Takayuki Hattori: "Molecular Cloning of a cDNA Encoding Human TSC-22 (Transforming Growth Factor β1-Stimulated Clone 22) and Localization of the Gene at Chromosome 13q14." Acta Sholae Medicinalis Universitatis in Gifu. 45巻. 201-208 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Nobuhisa Mizuki: "Nucleotide Sequence Analysis of the HLA Class I Region Spanning the 237 kb Segment Around the HLA-B and -C Genes." Genomics. 42巻. 55-66 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Toyoaki Tenzen: "Precise Switching of DNA Replication Timing in the GC Content Transition Area in the Human Major Histocompatibility Complex." Molecular and Cellular Biology. 17巻. 4043-4050 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Masamichi Ishiai: "Isolation of Human and Fission Yeast Gene Homologues of the Budding Yeast Origin Recognition Complex Subunit Orc5 : Human Homologue (ORC5L) Maps to 7q22." Genomics. (印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] Toshihiko Eki: "Mapping of the Human Genes Encoding Cyclin H (CCNH) and the CDK-activating Kinase (CAK) Assembly Factor NAT1 to Chromosome Bands 5q13.3-q14 and 14q23, Respectively" Genomics. (印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] Masashi Kimura: "Assignment of the STK6 to Human Chromosome 20q13.2-13.3 and a Pseudogene to 1q41-42." Cytogenetics and Cell Genetics. (印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] Chee-Gun Lee: "Molecular Analysis of the cDNA and genomic DNA encoding Mouse RNA Helicase A." Genomics. (印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] Sachiko Kitanaka: "Isolation of human 25-hydroxyvitamin D3 1α-hydroxylase gene and its mutations causing vitamin D-dependent rickets type I." New England Journal of Medicine. (印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] Katsuzumi Okumura: "Temporal Order of DNA Replication of Imprinted IGF2 Gene and Non-imprinted TNF and RFC5 Genes in Human Lymphocytes." Bulletin of the Faculty of Bioresources Mie University. (印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] Yasushi Taniguchi: "Nucleotide Sequence of the Ring3 Gene in the Class II Region of the Mouse MHC and Its Abundant Expression in Testicular Germ Cells." Genomics. (印刷中).

    • Related Report
      1997 Annual Research Report
  • [Publications] 奥村克純: "FISHによる動物細胞ゲノムの核内構造解析の現状." 生化学. 65巻. 41-46 (1997)

    • Related Report
      1997 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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