| Project/Area Number |
06454194
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Experimental pathology
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| Research Institution | Oita Medical University |
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Principal Investigator |
HIGUCHI Yasunori Oita Medical Univ.Pathology, assist.professor, 医学部, 助教授 (60040284)
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| Co-Investigator(Kenkyū-buntansha) |
SETOGUCHI Mihoko Oita Medical Univ.Pathology, assistant, 医学部, 助手 (20236110)
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| Project Period (FY) |
1993 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥7,200,000 (Direct Cost: ¥7,200,000)
Fiscal Year 1995: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1994: ¥5,300,000 (Direct Cost: ¥5,300,000)
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| Keywords | Transgene / Osteopotin / Metallothionein / Cytomegalovirus / U2alpha globulin / alpha1-antitrypsin / osteopontin / metallothionein / transgene / α1-antitripsin |
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| Research Abstract |
1.Establishment of transgenic mice (TM) with metallothionein (MT) promoter-driven mouse osteopontin (OP) gene and their analysis : A MT promotor-OP fusiongene was constructed, and everal founder mice were produced. However, use of the TM was impossible because of few number of litter mates and killing of new born TM by mother. Speciemens and samples from a few founder mice were subjected to biochemical and immunohistochemical analysis. Expression of transgenic RNA was abundant in the liver in TM on the basal diet. After maintenance with water containing ZnSO4 (50mM) for 4 days, expression of transgenic RNA was strong in the small intestine. Immunohistochemical analysis demonstrated OP expression in these organs in TM.Levels of OP in sera from TM after maintenance with water containing ZnSO4 (50mM) for 4 days were significantly higher than these animals maintained with normal water and nontransgenic mice. Founder mice were healthy and one of them is alive 2 years after birth. Autopsy ex
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amination of TM revealed no abnormal organs. 2.Establishment of trnansgenic mice (TM) with cytomegalovirus (CV) or U2alpha globulin (U2alpha) promoter-driven OP gene and their analysis : CV or U2alpha promoter-mouse osteopontin (OP) fusiongene was constructed, and a few founder mice were produced. However, we were anable to expand these TM.Speciemens and samples from a few founder mice were subjected to biochemical and immunohistochemical analysis. Northern blot and PAP analysis demonstrated OP in kidney in CVTM and in the liver in U2alpha TM.Levels of OP in sera from TM were significantly higher than nontransgenic mice. 3.Establishment of TM with alpha1-antitrypsin (alpha1AT) promoter-driven OP gene and their analysis : A alpha1AT-OP fusiongene was constructed, and several founder mice were produced in C57BL/6N/DBA/2F1 mice. Expansion of alpha1AT TM was easy. Backcross of heterozygote mouse to C57BL/6N to get TM of this backgrownd is now underway, reaching 6N.Northern blot and PAP analysis demonstrated OP in the liver in alpha1AT TM.Levels of OP in sera from TM were significantly higher than nontransgenic mice. Less
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