1995 Fiscal Year Final Research Report Summary
MOLECULAR CLONING OF THE HUMAN SPERM-ACTIVATING PROTEIN GENE AND THE LARGE SCALE PRODUCTION OF SPERMACTIVATING PROTEIN BY GENETIC ENGINEERING TECHNIQUE
| Project/Area Number |
06671680
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Obstetrics and gynecology
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| Research Institution | Dokkyo University |
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Principal Investigator |
OHTAKE Hideki Dokkyo Univ., Sch.of Med.Associate Professor, 医学部, 助教授 (00049214)
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| Project Period (FY) |
1994 – 1995
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| Keywords | Sperm-activating protein / Sperm-activating protein gone / Human ovary / Human spermatozoa / Sperm motility / Sperm activation / Molecular cloning |
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| 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.
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[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
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[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
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[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
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[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
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[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
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[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
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[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
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[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
