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MOLECULAR CLONING OF THE HUMAN SPERM-ACTIVATING PROTEIN GENE AND THE LARGE SCALE PRODUCTION OF SPERMACTIVATING PROTEIN BY GENETIC ENGINEERING TECHNIQUE

Research Project

Project/Area Number 06671680
Research Category

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

Allocation TypeSingle-year Grants
Research Field Obstetrics and gynecology
Research InstitutionDokkyo University

Principal Investigator

OHTAKE Hideki  Dokkyo Univ., Sch.of Med.Associate Professor, 医学部, 助教授 (00049214)

Project Period (FY) 1994 – 1995
Project Status Completed (Fiscal Year 1995)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1995: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
KeywordsSperm-activating protein / Sperm-activating protein gone / Human ovary / Human spermatozoa / Sperm motility / Sperm activation / Molecular cloning / 精子活性化蛋白質 / 精子活性化蛋白質遺伝子 / ニシン
Research Abstract

Recently, we purified sperm-activating proteins (HSAPs) from ripe unfertilized herring eggs and cloned a HSAP cDNA clone from a herring ovary cDNA library. Using the HSAP cDNA as a probe, the human ovary cDNA library was screened and cloned 8 positive clones which were thought to be homologous to the HSAP cDNA.The partial sequences (about 300 base pairs) of the cDNAs were determined and compared them with those in DNA sequence data bases such as the GenBank and the EMBL bank. The cDNA sequence of one clone is homologous to one of the cDNA fragments (about 300 bp) screend from human lymphoblastoma cDNA library by French scientists. But, the full lenght of the cDNA is not reported, therefore, the function of the gene product is not predicted at present. The cDNAs of the other clones were derived from unknown genes. To determine whether these genes are expressed only in ovary, each cDNA clons has been analyzing on mRNA species obtained from several human organs or tissues by Northern blot hybridization technique. The results revealed that at least one cDNA clone was expressed only in ovary. The analyzes of the other clones are currently underway.
To produce the HSAP gene product by genetic engineering, I tried to screen the full length cDNA clone, but failed to it. If the full length cDNA clone is obtained, the insert DNA is ligated to the Baculovirus expression vector and transfected to Sf9 insect culture cells. The recombinant proteins thus obtained will be examined whether the proteins have the capacity for activating the motility of the human spermatozoa.

Report

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

    (13 results)

All Other

All Publications (13 results)

  • [Publications] Oda,S,Igarashi,Y,Ohtake,H,Sakai,K,Shimizu,N.&Morisawa,M.: "Spemu activeting piotein from Unfertilzod eggs of the Pacific Perring ,Clupeapalasii" Develop.Growth Differ.37. 257-261 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tadokoro,N,Koibuchi,N.Ohtake,H,Kowatsu,T,Kato,Y,Yamaoka,S.&Matsugahs,T.: "Expression of prolactin gene in hcman deciduo during pregnancy studied by in situ hybidization histochemisty." Findocrire J.42. 537-543 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Koibuchi,N,Komo,R.,Matsuzaki,S.,Ohtake,H,Niwa,A.&Yamaoka.S.: "Eocalization of D,amirco acid oxidase m RNA in the mousl kidnoy and the effact of tostostsone tro atiment" Histochem Cell Biol. 104. 349-355 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tadokoro,N.,Koibuchi,N.,Ohtake,H.,Kawatsu,T,Ohkawa,H,Yamaoka,S.&Kumasaka,T.: "Localization of prolaction and ito receptor,massenger RNA in the human decidua" Experientia. 51. 1216-1219 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Oda, S., Igarashi, Y., Ohtake, H., Sakai, K., Shimizu, N.& Morisawa, M: "Sperm-activating proteins from unfertilized eggs of the Pacific herring, Clupea palasii" Development Growth and Differentiation. 37. 257-261 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tadakor, N., Koibuchi, N., Ohtake, H., Kawatsu, T., Kato, Y., Yamaoka, S.& Matsuzaki, T.: "Expression of prolactin gene in human decidua during pregnancy studied by in situ hybridization histochemistry" Endocrine J.42. 537-543 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Koibuchi, N., Nonno, R., Matsuzaki, T., Ohtake, H., , Niwa, A.Yamaoka, S.: "Localization of D-amino acid oxidase mRNA in the mouse kidney and the effect of testosterone treatment" Histocem.Cell Biol.104. 349-355 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Tadokoro, N., Koibuchi, N., Ohtake, H., Kawatsu, T., Ohkawa, H., Yamaoka, S.& Kumasaka, T.: "Localization of prolactin and its receptor messenger RNA in the human decidua" Experientia. 51. 1216-1219 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1995 Final Research Report Summary
  • [Publications] Oda, S., Igavashi, Y., Ohtake, H., Sahai, K., Shimizu, N., & Morisawa, M.: "Sperm activating proteins from unfertilized eggs of the Pacific horring, Clupea pallasii" Develop. Growth Differ. 37. 257-261 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Tadokoro, N., Koibuchi, N., Ohtake, H., Kawati, T., Kato, Y., Yamaoka, S., & Kumasaka, T.: "Expression of Prolactin Gene in Human desidua during Prognancy Studied by in Oitu Hybridigation Histochemistry." Endocrine J.42. 537-543 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Koibushi, N., Konno, R., Matsuzaki, S., Ohtake, H., Niwa, A., Yamashe, S.: "Localization of D-amino acid Oxidase mRNA in the mouse kidney and the effect of testosterone treatment" Aistochem Cell Biol. 104. 349-355 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] Tadokoro, N., Koibuchi, N., Ohtake, H., Kawatsu, T., Ohkawa, H., Kato, Y., Yamashe S., & Kumasaka, T.: "Localization of prolactin and its receptor messenger RNA in the human cleciclua" Experientia. 51. 1216-1219 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] S.Oda,Y.Igarashi,K.Sakai,M.Morisawa,H.Ohtake & N.Shimizu: "Sperm activating proteins from unfertiliged egg of the Pacific herring, Clupea pallasi." Development,Growth & Differentiation. 37(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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