| 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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| Project Status |
Completed (Fiscal Year 1995)
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| 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)
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| Keywords | Sperm-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 |
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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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