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Development of transgenic mice expressing human complement modulator

Research Project

Project/Area Number 06454718
Research Category

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

Allocation TypeSingle-year Grants
Research Field Laboratory animal science
Research InstitutionOsaka University Research Institute for Micobial Diseases

Principal Investigator

OKABE Masaru  Research Institute for Microbial Diseases, Associate Professor, 微生物病研究所, 助教授 (30089875)

Co-Investigator(Kenkyū-buntansha) SEYA Tsukasa  Osaka Research Center for Adult Disease, Chief research, 第六部, 室長
NISHIMUNE Yoshitake  Research Institute for Microbial Diseases, Professor, 微生物病研究所, 教授 (80029793)
Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥5,100,000 (Direct Cost: ¥5,100,000)
Keywordscomplement / transgenic mouse / virus receptor / reproductive immunology / ウイルスセプター
Research Abstract

Production of transgenic mice which express CD46
The construct to make transgenic mice were checked using CHO cells in vitro. The checked construct which is consisted with SV40 SR alpha promoter and CD46 were used in transgenic mouse experiment. However, all the transgenic mice produced by this construct did not express CD46 on their cell surface. On the course of finding the cause of this silent expression, it was found that when the cytosolic tail of CD46 were eliminated, the rest of the CD46 were expressed. Further experiment using transgenic mice also revealed that the origin of the silencer effect is derived form the nontranslated 3' region of the CD46 gene. By making the transgenic mice lacking CD46 3' untranslated region, we could produce the mice which express the CD46 on the cell surface.
Cloning of CD46 genomic gene.
The expression pattern of the above mentioned transgenic mice were limited to muscle, pancreas, lung and some more organs, and not strongly on the blood cells. The virus infection test was performed, but until now a very concrete evidence of the infection is not obtained. Therefore, to express CD46 as in the natural CD46 expressing cells, we cloned the genomic CD46 from human genomic library. At the present moment, we are now producing CD46 transgenic mice driven under CD46 promoter.
The role of CD46 in fertilization.
CD46 was shown to act in the complement regulation and also in sperm-egg fusion. In the seminal fluid, there are considerable amount of CD46. The CD46 in the seminal fluid was shown to exist on prostasome membrane and has an effect on complement regulation. The role of the prostasome CD46 was suggested to protecting sperm from complement attack in the vagina and uterus.

Report

(3 results)
  • 1995 Annual Research Report   Final Research Report Summary
  • 1994 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] K. Iwata, et al.,: "Diversity of sites for measles virus binding and for inactivation of complement C36 and C46 on membrane cotactor protein CD46;" J. Biol. Chem. 270. 15148-15152 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M. Kitamura, et al.,: "Membrane cotactor protein (CD46) in seminal plasma is a prostasome-beand from with compiement regulatory activity and measles virus meatralizing activity;" Immunology. 84. 626-632 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M. Ikawa, et al.,: "Greentlaorescent protein as a marker intransgenic mice;" Develop. Growth Differ.,. 37. 455-459 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M. Ikawa, et al.,: "Arapid and non-invasive selection of transgenic embryos betone implantation using green flucrescent protein(GFP);" FEBS Lett.,. 375. 125-128 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] X. Ying, et al.,: "Improvement of fusing ability of human Sperm to Zona-Free hamster eggs by conditioned media;" Biol. Pharm. Ball.,. 18. 5-8 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] D. Watanabe, et al.,: "Characterization of the testis-spocific gene ‘calmegin' Promoter seguence and its activity defined by transgenic mouse experiments;" FEBS Lett.,. 368. 509-512 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Kitamura, M.Namiki, K.Matsumiya, K.Tanaka, M.Matumoto, T.Hara, H.Kiyohara, M.Okabe, A.Okuyama and T.Seya: "Membrane cofactor protein (CD46) in seminal plasma is a prostasome-bound form with complement regulatory activity and measles virus" Immunology. 84. 626-632 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] D.Watanabe, M.Okabe, N.Hamajima, T.Morita, Y.Nishina and Y.Nishimune: "Characterization of the testis-specific gene, calmegin' promoter sequence and its activity defined by transgenic mouse experiments" FEBS Lett.368. 509-512 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Ikawa, K.Kominami, Y.Yoshimura, K.Tanaka, Y.Nishimune and M.Okabe: "A rapid and non-invasive selection of transgenic embryos before implantation using green fluorescent protein (GFP)" FEBS Lett.375. 125-128 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Ikawa, X.Ying, K.Tanaka, K.Ohashi, F.Saji and M.Okabe: "Improvement of fusing ability of human Sperm to zona-Free hamster eggs by conditioned media" Biol.Pharm.Bull.18. 5-8 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] M.Ikawa, K.Kominami, Y.Yoshimura, K.Tanaka, Y.Nishimune and M.Okabe: "Green fluorescent protein as a maker in transgenic mice" Develop.Growth Differ.37. 455-459 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Iwata, T.Seya, Y.Yanagi, J.Pesando, P.Johnson, M.Okabe, S.Ueda, H.Ariga and S.Nagasawa: "Diversity of sites for measles virus binding and for inactivation of complement C3b and C4b on membrane cofactor protein CD46" J.Biol.Chem.270. 15148-15152 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Ohashi, F.Saji, A.Wakimoto, T.Tsutsui, T.Nakazawa, M.Okabe, T.Mimura and O.Tanizawa: "Selection of acrosome-reacted sperm with MH61-Immunobeads" Am.Soc.Androl.15. 78-82 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] K.Iwata et al.,: "Diversity of sites for measles virus binding and Frinactivation of complement C3b and C4b on membrane cofactor protein CD46 ;" J.Biol.Chem.270. 15148-15152 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] M.Kitamura et al.: "Membrane cofactor protein (CD46)in seminal plasma is a prostasome taiurd form with complement regulatory activity and measles virus neutralizing activity." Immurology. 84. 626-632 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] M.Ikawa et al.,: "Green fluorescent protein as a marker in transgenic mice" Develop.Growth Oiffer.37. 455-459 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] M.Ikawa et al.,: "A rapid and non-invasive selection of transgenic embryos. before implantation using green fluorescent protein(GFP)" FEBS Lett,. 375. 125-128 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] X.Ying et al.,: "Improvement of fusing ability of human Spenu to Zena-free hainstr eggs by conditioneal metra" Bicel.Pharm-Bull.18. 5-8 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] D.Watanabe et al.,: "characterization of the testis-specific gene ‘calmegin'promoter sebuene and its activity defined by transgenic mause expeniments" FEBS Lett.,. 368. 509-512 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] K.Iwata et al.,: "Diversity of sites for measles virus binding and for inactivation of complement C3b and C4b on membrane cofactor protein(CD46,MCP)" J.Biol.Chem.(in press). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] M.Kitamura et al.,: "Membrane cofactor protein(MCP,CD46)in seainal plasma in a prostasome-bound form with complement regulatory activity and measks virus neutralizing activity." Immunology. 84(in press). (1995)

    • Related Report
      1994 Annual Research Report

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Published: 1994-04-01   Modified: 2016-04-21  

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