Establishment of immunochemical determination of human cholesterol 7 alpha-hydroxylase
Project/Area Number |
07671413
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Research Category |
Grant-in-Aid for Scientific Research (C)
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Allocation Type | Single-year Grants |
Section | 一般 |
Research Field |
Digestive surgery
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Research Institution | MIYAZAKI MEDICAL COLLEGE |
Principal Investigator |
MAEDA Yorio MIYAZAKI MEDICAL COLLEGE,SURGERY I,ASSISTANT PROFESSOR, 医学部, 講師 (60117463)
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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 |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1996: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1995: ¥1,900,000 (Direct Cost: ¥1,900,000)
|
Keywords | cholestrol 7alpha-hydroxylase / monoclonal antibody / fusion protein immunoblotting / コレステロール7α水酸化酵素 / fusion protein |
Research Abstract |
A fusion protein was prepared from an in-frame fusion gene containing the cDNA for human cholesterol 7alpha-hydroxylase near the 3' terminus of the lacZ gene of Escherichia coli and monoclonal antibody angainst human cholesterol 7alpha-hydroxylase was established. Using this antibody, immunochemical determination was established. To clarify the enzyme level of cholesterol 7alpha-hydroxylase during liver regeneration, we determined the enzyme level after hepatectomy in the rats. We also determined mRNA level and enzyme activity. 12hrs after hepatectomy, the level of mRNA,enzyme activity and enzyme protein mass decreased in parallel and gradually returned to normal level. These results suggest that this enzyme protein decreased during cell division and is under pretranslational regulation. Now we are preparing the manuscript concerning the species difference of gene expression of cholesterol 7alpha-hydroxylase after bile duct ligation in the rats and rabbits. In the course of pursuing the project, we have investigated the enzymes, 3beta-hydroxy-DELTA^5-C_<27>-steroid dehydrogenase and 25-hydroxyvitamin D_3 1alpha-hydroxylase, which are related to the cholesterol 7alpha-hydroxylase and published in the journals.
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Report
(3 results)
Research Products
(6 results)
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[Publications] Maeda, Y., Junji, N., Kinowaki, M., Kuroki, N., Shushi, H.and Setoguchi, T.: "Bile acid metabolism during liver regeneration in the rats." Surgical Therapy. 77(2). 236-237 (1997)
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[Publications] Maeda, Y., Ando, Y., Nagatomo, H., Uchiyama, F., Higashi, S.and Setoguchi, T.: "Sex differences in serum 7alpha-hydroxycholesterol levels in the rat reflect hepatic activity of 3beta-hydroxy-DELTA^5-C_<27>-steroid dehydrogenase and cholesterol 7alpha-hydroxylase." Journal of Gastroenterology. 32. 502-506 (1997)
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[Publications] Eto, T.-A., Nakamura, Y., Taniguchi, T., Miyamoto, K., Nagatomo, J., Maeda, Y., Higashi, S., Okuda, K.-I.and Setoguchi, T.: "Assay of 25-hydroxyvitamin D_3 1alpha-hydroxylase in rat kidney mitochondria." Analytical Biochemistry. 258. 53-58 (1998)
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