| Project/Area Number |
13671666
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Urology
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| Research Institution | Osaka City University |
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Principal Investigator |
SUGIMURA Kazunobu Osaka City University, Graduate School of Medicine, Associate Professor, 大学院・医学研究科, 助教授 (90187659)
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| Project Period (FY) |
2001 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2003: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2002: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2001: ¥700,000 (Direct Cost: ¥700,000)
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| Keywords | Polycystic Kidney / DPT / Differential display / sulfotransferase / SULT1C2 / recombinant protein / のう胞腎 / PKD |
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| Research Abstract |
Background. The pathogenesis of polycystic kidney disease (PKD) remains unclear in spite of the identification of the genes responsible for hereditary PKDs. In this study, we investigated the alteration of gene expressions in an acquired PKD model induced by 2-amino-4,5-diphenylthiazole (DPT) using the differential display method.
Methods. Kidney mRNA from a Sprague-Dawley rat fed with 1% DPT for 4 days and from a control rat was compared by RT-PCR differential display method. Differentially expressed bands were re-amplified and subcloned. Using these subclones as probes, the changes in gene expressions were confirmed by Northern blot analysis. Subsequently, mouse kidney cDNA library was screened.
Results. The isolated 1.5-kb cDNA contained an open reading frame encoding 296 amino acids which shared 94.3% identity with rat SULT1C2 sulfotransferase, and was considered to be its mouse orthologue (GenBank Accession No. AY005469). Mouse SULT1C2 mRNA was abundant in the kidney and stomach among normal mouse tissues. The expression of SULT1C2 mRNA was decreased in the rat kidney after DPT feeding but not in the stomach. Mouse SULT 102 was expressed successfully using pET plasmid vector and F.coil The recombinant 34kDa protein was capable of catalyzing the sulfation of p-nitrophenol at Km of 3.1 mM, by utilizing 3'-phosphoadenosine 5'-phosphosulfate (PAPS) as the sulfate donor.
Conclusions. Although the physiological substrate and function of SULT1C2 have yet to be elucidated, its down-regulation could be involved in the cystic changes of tubules by decreasing the sulfation of the tubular basement membrane components.
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