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1992 Fiscal Year Final Research Report Summary

Molecular cloning and basic study of bone inducing factors for their clinical use.

Research Project

Project/Area Number 03670937
Research Category

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

Allocation TypeSingle-year Grants
Research Field 外科・放射線系歯学
Research InstitutionTOKYO MEDICAL AND DENTAL UNIVERSITY

Principal Investigator

IWAKI Hiroshi  Tokyo Med.& Dent.Univ.,1st Dept.of Oral & Maxillofacial Surg., Faculty of Dentistry, Research Associate, 歯学部, 助手 (70107308)

Co-Investigator(Kenkyū-buntansha) MOMOSE Fumio  Tokyo Med.& Dent.Univ., 1st Dept.of Oral & Maxillofacial Surg., Faculty of Denti, 歯学部, 助手 (60157849)
OIDA Shinichiro  Tokyo Med.& Dent.Univ., Biochemistry, Faculty of Dentistry Research Associate, 歯学部, 助手 (10114745)
Project Period (FY) 1991 – 1992
KeywordsBone Morphogenetic Protein(BMP) / Polymerase Chain Reaction(PCR) / Recombinant Protein
Research Abstract

To understand the mechanism of biological actions of bone inducing factors, we attempted to clone the genes for these factors.
According to the human BMP(Bone Morphogenetic Protein)-2/4 and mouse BMP-6(Vgr-1) nucleotide sequences, oligonucleotide primers were prepared. Messenger RNAs were isolated from neonatal mouse tissue and human osteosarcoma cell line MG63, and used for the synthesis of complementary DNAs (cDNAs) with reverse transcriptase. By PCR(Polymerase Chain Reaction) using these primers and cDNA templates, we have cloned cDNAs for mouse BMP-2, mouse BMP-6, and human BMP-4.
To isolate full length of BMP cDNAs, various cDNA libraries were screened with the cDNA probes cloned by PCR. A positive clone detected in the screening of a human placenta cDNA library was subcloned into a plasmid vector and subjected to nucleotide sequencing. The result was that the positive clone was found to be a full length cDNA for human BMP-4. Using the same method, an almost full length human BMP-6 cDNA was also isolated from a human lung small cell carcinoma(NCL-H69) cDNA library.
Furthermore, we attempted to synthesize the human BMP-4 recombinant protein. The human BMP-4 cDNA was transferred into two derivatives of the E.coli expression vector, and a beta - Galactosidase-human BMP-4 fusion protein and a Glutathione-S- transferase-human BMP-4 fusion protein were obtained. Osteoinductive activities of these fusion proteins are now under investigation.

  • Research Products

    (6 results)

All Other

All Publications (6 results)

  • [Publications] Shinichiro OIDA: "PCR amplification and cloning of a mouse bone morphogenetic protein(BMP)cDNA" Japanese Journal of Oral Biology. 34. 612-615 (1992)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 丸岡 豊: "PCR法によるヒト骨形成タンパク質-2(hBMP-2)のcDNAの単離・増幅" 歯科基礎医学学会雑誌. 35. 64-74 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 二藤 彰: "骨形成に関与する細胞成長因子についての基礎的研究第1報骨形成タンパク質cDNAのPCR法によるクローニング" 日本口腔外科学会雑誌. 39. 103-109 (1993)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Shinichiro Oida: "PCR amplification and cloning of a mouse bone morphogenetic protein (BMP) cDNA." Jpn.J.Oral.Biol.34. 612-615 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yutaka Maruoka: "Cloning of human bone morphogenetic protein-2 (hBMP-2) cDNA by PCR method." Jpn.J.Oral.Biol.35. 64-74 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] NITOU.Akira: "A study on growth factors regulating bone formation Part1 cDNA cloning of murine bone morphogenetic protein by polymerase chain reaction." Jpn.J.Oral Maxillofac.Surg.39. 103-109 (1993)

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

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Published: 1994-03-24  

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