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1992 Fiscal Year Final Research Report Summary

Improvement of high-resolution chromosome banding methods, and its application to human gene mapping.

Research Project

Project/Area Number 02454492
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field Human genetics
Research InstitutionTokyo Medical and Dental University

Principal Investigator

IKEUCHI Tatsuro  Med.Res.Inst., Tokyo Med.Dent.Univ., Assoc.Prof., 難治疾患研究所, 助教授 (90041839)

Co-Investigator(Kenkyū-buntansha) YOSHIDA Mitsuaki  Med.Res.Inst., Tokyo Med.Dent.Univ., Assist.Prof., 難治疾患研究所, 助手 (60182789)
SAITO Fumiko  Med.Res.Inst., Tokyo Med.Dent.Univ., Assist.Prof., 難治疾患研究所, 助手 (10158917)
Project Period (FY) 1990 – 1992
KeywordsHigh-resolution chromosome banding / Ethidium bromide method / Cell synchronization method / Hereditary disease loci / Lymphoblastoid cell lines / Fluorescent in situ hybridization / DNA segment clones / Human No.21 chromosome
Research Abstract

1. Various established methods for high-resolution chromosome banding were re-evaluated and in part improved for their application to different cell culture systems.
(1) The ethidium bromide (EB) method was found to be applicable, by improving the dose and duration of treatment with EB and Colcemid, to the well-grown tumor cells in culture.
(2) The method of cell synchronization with methotrexate (MTX) and subsequent BrdU treatment which was applied to lymphocyte cultures from the dog (Canis familiaris) yielded sufficient results for high-resolution replication R-banding.
(3) A reliable method to obtain high-resolution R-banded chromosomes from lymphoblastoid cell lines (LCL) was established by combination of EB addition (1-1.5 hr) and excess thymidine-induced cell synchronization followed by the BrdU treatment (6.5-7 hrs).
2. By applying the EB method to peripheral lymphocyte cultures, the break points of various structural chromosome abnormalities were precisely defined, leading to the regional mapping of the following disease loci : Down syndrome-related region 21q22.2->qter), HMC syndrome (1q31.2 or 7p15.1-p15.3), neurofibromatosis type 2 (22q12.2).
3. Fluorescent in situ hybridization was performed for molecular cytogenetic characterization of some structural chromosome abnormalities (rings, pseudodicentrics, etc.) by using DNA probes of repeated sequences (telomere'DNA, rDNA alpha satellite DNA and DNA segments containing Alu repeats). Furthermore, precise regional mapping of a total of 20 DNA segments derived from human No.21 chromosomes were performed by FISH analysis and high-resolution R-banding. They include 13 NotI- and 7 SfiI-linking clones.

  • Research Products

    (29 results)

All Other

All Publications (29 results)

  • [Publications] Ikeuchi T,et al.: "Ring chromosome 21 transmitted from mother to daughter its stability in a lymphoblastoid cell line." Annales de Genetique.33. 32-35 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Korenberg JR,et al.: "Molecular definition of a refgion of chromosome 21 that causes features of the Down syndrome phenotype." American Journal of Human Genetics.47. 236-246 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Poulsen BS,: "Banding suudies Canis familiaris.I.Replication patterns in Karyotypes from lymphsocyte cultures." Cytobios. 62. 161-165 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ikeuchi,T,et al.: "Refined determination of breakpoints of the translocation t(1;7)assocaited with signs of HMC syndroms." Japanese Journal of Human Genetics.36. 155-158 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Asano,T.,et al.: "Partial 18qtrisomy and 18p monosomy resulting from a maternal pericentric inversion,inv(18)(pll.2q21.3)." Japanese Journal of Human Genetics.36. 257-265 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Kojima,A.,et al.: "Y/6 chromosome translocation in a male with triple primary cancers involving the breast." Journal of Cancer Research and Clinical Oncology.117. 479-483 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yosida,M.A.,et al.: "Chromosome changes in desmoid tumors developed in patients with familial adenomatous polyposis." Japanese Journal of Cancer Research.82. 916-921 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Arai,E.,et al.: "Constitutional translocation t(4;22)(q12;q12.2)associated with neurofibromatosis type 2." American Journal of Medical Genetics. 44. 163-167 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hirabayashi, Y.,et al.: "Chromosomal rearrangements at 12q13 in two cases of chondrosarcomas." Cancer Genetics and Cytogenetics. 60. 35-40 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yin,M.,et al.: "Cytogenetic studies of human malignant melanoma cell lines." The Bulletion of Tokyo Medical and Dental University. 39. 43-54 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Arai,E.,et al.: "Mapping the breakpoint of a constitutional translocation on chromosome 22 in a patient with NF2." Genes Chromosomed and Cancer. in press. (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 池内達郎: "高精度バンディングによる遺伝子マッピング" 代謝(臨時増刊号)「先端のバイオサイエンス」. 27. 383-391 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 吉田光明、池内達郎: "固形腫瘍の初代培養と染色体解析" 組織培養. 17. 159-163 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 池内達郎: "ヒト染色体の微細構造." 生物の科学・遺伝(ヒトの遺伝). 別冊No.5. 23-33 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 池内達郎: "「ヒト培養細胞および染色体診断マニュアル」(武部啓・編)、講談社サイエンティフィク" High-resolution banding(高精度分染), 57-64 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 池内達郎、吉田光明: "「ヒト培養細胞および染色体診断マニュアル」(武部啓・編)、講談社サイエンティフィク" コンピュータによる染色体解析, 160-167 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ikeuchi, T., et al.: "Ring chromosome 21 transmitted from mother to daughter : its stability in a lymphoblastoid cell lines." Annal. Genet.33. 32-35 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Korenberg, J.R., et al.: "Molecular definition of a region of chromosome 21 that casuses features of the Down syndrome phenotype." Amer.J.Human Genet.47. 236-246 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Poulsen, B.S.: "Banding studies in Canis familiaris. I.Replication patterns in karyotypes from lymphocyte cultures." Cytobios. 62. 161-165 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ikeuchi, T., et al.: "Cytogenetic studies on colon carcinomas and adenomas from patients with familial polyposis coli." Hereditary Colorectal Cancer (J.Utsunomiya & H.T.Lynch, eds.). 533-540 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ikeuchi, T., et al.: "Refined determination of breakpoints of the t(1 ; 7) associated with sings of HMC syndrome." Japan. J.Human Genet.36. 155-158 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Asano, T., et al.: "Partial 18q trisomy and 18p monosomy resting from a maternal pericentric inversion, inv (18)(p11.2q21.3)." Japan. J.Human Genet.36. 257-265 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Kojima, A., et al.: "Y/6 chromosome translocation in a male with triple primary cancers involving the breast." J.Cancer Res.Clin., Oncol.117. 479-483 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoshida, M.A., et al.: "Chromosome changes in desmoid tumors developed in patients with familial adenomatous polyposis." Japan. J.Cancer Res.82. 916-921 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Sugawara, I., et al.: "Characterization of an etoposide-resistant human K562 cell line, K/eto." Japan, J.Cancer Res.82. 1035-1043 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Arai, e., et al.: "Constitutional translocation t(4 ; 22) (q12 ; q12.2) associated with neurofibromatosis type 2." Amer. J.Med. Genet.44. 163-167 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hirabayashi, Y., et al.: "Chromosomal rearrangements at 12q13 in two cases of chondrosarcomas." Cancer Genet. Cytogenet.60. 35-40 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yin, M., et al.: "Cytogenetic studies of human malignant melanoma cell lines." Bull.Tokyo Med.Dent.Univ.39. 43-54 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Arai, E., et al.: "Mapping the breakpoint of a constitutional translocation on chromosome 22 in a patient with NF2." Genes Chrom. Cancer. in press. (1993)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1995-03-27  

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