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Development of DNA chips for ribosomal protein genes

Research Project

Project/Area Number 11694300
Research Category

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

Allocation TypeSingle-year Grants
Section一般
Research Field General medical chemistry
Research InstitutionUniversity of the Ryukyus

Principal Investigator

KENMOCHI Naoya  University of the Ryukyus, Biochemistry, Instructor, 医学部, 助手 (00133124)

Co-Investigator(Kenkyū-buntansha) TANAKA Tatsuo  University of the Ryukyus, Biochemistry, Professor, 医学部, 教授 (70018688)
Project Period (FY) 1999 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥4,300,000 (Direct Cost: ¥4,300,000)
Fiscal Year 2000: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1999: ¥2,400,000 (Direct Cost: ¥2,400,000)
Keywordsribosome / DNA chip / human disease / ribosomal protein / microarray / gene expression / mutation screening
Research Abstract

In this study, we have developed DNA chips for human ribosomal protein (RP) genes, in collaboration with Dr.Metspalu at Tartu University in Estonia, to examine the possible involvement of RP gene defects in human disorders. We prepared following two types of DNA chips ; one for hybridization and the other for primer extension on the chip.
1. RPex (Ribosomal Protein gene expression) chip
PCR products (〜250 bp) of cDNAs for 40S ribosomal small subunit proteins were immobilized onto an epoxy-silanized glass surface. The gene expression at the mRNA level was measured by hybridization of fluorescently labeled DNA probes prepared from total mRNA of either wild-type HeLa cells or heat-shock HeLa cells. Although there were no significant differences of the RP gene expression in these two cells, we found the differences in RPS17, RPS18, RPS24, and RPS27 when the expression was compared between the HeLa and placental RNAs. This suggests that the changes of the RP gene expression level are involved in the carcinogenesis.
2. RPmu (Ribosomal Protein gene mutation) chip
Oligoncleotides (25 mer) specific to the small subunit protein genes were immobilized via 5'terminal amino group on an epoxy-silanized glass surface. After hybridization of target DNA to the array, target dependent oligonucleotide extension by a DNA polymerase is used to incorporate fluorescently labeled dideoxy terminators to the primers. More than 80% of the primers worked properly and precise single-base extensions were occurred on the array.
These two chips shotuld be useful for studying the RP gene defects or mutations in human disorders.

Report

(3 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • Research Products

    (28 results)

All Other

All Publications (28 results)

  • [Publications] Higa.S: "Gene organization and sequence of the region containing the ribosomal protein gene RPL13A and RPS11 in the human genome and conserved features in the mouse genome."Gene. 24. 371-377 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 剣持直哉: "リボソームの分子解剖:超分子システムの遺伝子解析."バイオサイエンスとインダストリー. 11. 781-784 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ion,A.: "Detailed mapping, mutation analysis, and intragenic polymorphism identification in candidate Noonan syndrome genes MYL2, DCN, EPS8, and RPL6."J.Med.Genet.. 37. 884-886 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurg,A.: "Arrayed Primer Extension : Solid phase four-color DNA resequencing and mutation detection technology"Genetic Testing. 4. 1-7 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tonisson,N.: "Unravelling Genetic Data by Arrayed Primer Extension."Clinical Chemistry and Laboratory Medicine. 38. 165-177 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Gasparini,P.: "High carrier frequency of the 35delG deafness mutation in European populations."Eur.J.Hum.Genet.. 8. 19-23 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kenmochi,N.: "The human ribosomal protein L6 gene in a critical region for Noonan syndrome."J.Hum.Genet.. 45. 290-293 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Uechi,T.: "A complete map of the human ribosomal protein genes : Assignment of 80 genes to the cytogenetic map and implications for human disorders."Genomics. 72(in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Metspalu,A.: "Microsystems Technology : A Powerful Tool for Biomolecular Studies"Birkhaeuser Verlag. 26 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tonisson,N.: "DNA Microarrayes : Biology and Technology"Bio Techniques Books. 16 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Higa S: "Gene organization and sequence of the region containing the ribosomal protein gene RPL13A and RPS11 in the human genome and conserved features in the mouse genome."Gene. 24. 371-377 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Metspalu A: "DNA resequencing, mutation detection and gene expression analysis by oligonucleotide microchips. In : "Microsystems Technology : A Powerful Tool for Biomolecular Studies" ed.by M.Kohler et al."Birkhaeuser Verlag. 371-397 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kenmochi N: "Molecular dissection of the ribosome gene organization of an elaborate molecular machinery."Bioscience and Industry. 11. 781-784 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ion A: "Detailed mapping, mutation analysis, and intragenic polymorphism identification in candidate Noonan syndrome genes MYL2, DCN, EPS8, and RPL6."J.Med.Genet.. 37. 884-886 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kurg A: "Arrayed Primer Extension : Solid phase four-color DNA resequencing and mutation detection technology."Genetic Testing. 4. 1-7 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tonisson N: "Unravelling Genetic Data by Arrayed Primer Extension."Clinical Chemistry and Laboratory Medicine. 38. 165-177 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Gasparini P: "High carrier frequency of the 35delG deafness mutation in European populations. Genetic Analysis Consortium of GJB2 35delG."Eur.J.Hum.Genet.. 8. 19-23 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Tonisson N: "Arrayed primer extension on the DNA chip-method and applications. In "DNA Microarrayes : Biology and Technology" Ed by M.Schena"Bio Techniques Books. 247-263 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kenmochi N: "The human ribosomal protein L6 gene in a critical region for Nooonan syndrome."J.Hum.Genet.. 45. 290-293 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Uechi T: "A complete map of the human ribosomal protein genes : Assignment of 80 genes to the cytogenetic map and implications for human disorders."Genomics. (in press).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 剣持直哉: "リボソームの分子解剖:超分子システムの遺伝子解析."バイオサイエンスとインダストリー. 11. 781-784 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kenmochi,N.: "The human ribosomal protein L6 gene in a critical region for Noonan syndrome."J.Hum.Genet.. 45. 290-293 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Uechi,T.: "A complete map of the human ribosomal protein genes : Assignment of 80 genes to the cytogenetic map and implications for human disorders."Genomics. 72(in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kurg,A.: "Arrayed Primer Extension : Solid phase four-color DNA resequencing and mutation detection technology"Genetic Testing. 4・1. 1-7 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Gasparini,P.: "High carrier frequency of the 35delG deafness mutation in European populations."Eur.J.Hum.Genet.. 8・1. 19-23 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tonisson,N.: "Unravelling Genetic Data by Arrayed Primer Extension."Clinical Chemistry and Laboratory Medicine. 38・2. 165-177 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tonisson,N.: "DNA Microarrayes : Biology and Technology"BioTechniques Books. 16 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Higa,S.,Yoshihama,M.,Tanaka,T.,Kenmochi,N.: "Gene organization and sequence of the region containing the ribosomal protein gene RPL13A and RPS11 in the human gemome and conserved features in the mouse genome."Gene. 24. 371-377 (1999)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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