| Project/Area Number |
07556124
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 試験 |
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| Research Field |
Applied veterinary science
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| Research Institution | KITASATO UNIVERSITY |
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Principal Investigator |
MORI Junichi Kitasato University, School of Veterinary Medicine & Animal Sciences, Ph.D., Professor, 獣医畜産学部, 教授 (90167685)
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| Co-Investigator(Kenkyū-buntansha) |
IHARA Takeshi Nippon Institute for Biological Science, D.V.M., Project Leader, 日本生物科学研究所, 研究員 (70150109)
INABA Toshio Osaka Prefecture University, College of Agriculture Ph.D., Associate Professor, 農学部, 助教授 (00137241)
KATO Yukio Gunma University, Institute for Molecular and Cellular Regulation, Ph.D., Associ, 生体調節研究所, 助教授 (30114177)
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| Project Period (FY) |
1995 – 1996
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| Project Status |
Completed (Fiscal Year 1996)
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| Budget Amount *help |
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1996: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | Gonadotropins / Recombinant DNA / Pig / Vaculovirus / Embryo transfer / Follicle stimulating hormone / Luteinizing hormone / バキュロウイルスベクター |
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| Research Abstract |
Production of biologically active recombinant porcine FSH (rpFSH) free from cognate pituitary glycoprotein hormone LH was performed. The rpFSH was synthesized by a baculovirus vector-insect cell system using two cDNAs ecoding glycoprotein alpha and FSH beta subunits. Its antigenicity was same as that of pFSH prepared from pituitary. Glycosylation of rpFSH was observed by treatment of tunicamycin but the molecular weight of each subunit was lower than that of pituitary-darived FSH (pFSH), due to the absence of trimming of terminal sugar and addition of sialic acid in insect cells. RpFSH was secreted in the culture media at about 1 mg/l and purified in 6 fractions, due to the heterogeneity of sugar group, by S-Sepharose and Con-A column chromatographies. The biological activity of rpFSH was examined in progesterone secretion from porcine granulosa cells and germinal vesicle break down (GVBD) of porcine cocytes. RpFSH showed adcquate activity of progesterone sacretion, though some fractions rich in sugar group showed lower activity than that of pFSH.RpFSH gave higher GVBD activity than that of pFSH at low concentration as 1 ng/ml. These results demonstrated that baculovirus vector-insect cell system provided biologically active rpFSH.Moreover, rpFSH induced ovulation of pregnant mare serum gonadotropin (PMSG) primed and hypophysectomized immature female rats, with associated increase in tissue-type plasminogen activator (tPA).
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