2005 Fiscal Year Final Research Report Summary
Analysis of cystic disease of kidney by making mice defective of Sulfotransfer gene (Sult1C2)
| Project/Area Number |
16591612
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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 |
YOSHIMURA Rikio Osaka City University, Graduate School of Medicine, Lecturer, 大学院・医学研究科, 講師 (50285293)
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| Co-Investigator(Kenkyū-buntansha) |
SUGIMURA Kazunobu Osaka City University, Graduate School of Medicine, Associate Professor, 大学院・医学研究科, 助教授 (90187659)
MORITA Takashi Osaka City University, Graduate School of Medicine, Professor, 大学院・医学研究科, 教授 (70150349)
YOSHIDA Kayo Osaka City University, Graduate School of Medicine, Lecturer, 大学院・医学研究科, 講師 (30311921)
YOSHIDA Shuhei Osaka City University, Graduate School of Medicine, Research Associate, 大学院・医学研究科, 助手 (20363997)
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| Project Period (FY) |
2004 – 2005
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| Keywords | Sulfotransferase / Sult1C2 gene / gene-deficient mice / Polycystic Kidney / Carcinogenesis |
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| Research Abstract |
Polycystic kidney is induced by feeding 1%2-amino-4,5-diphenylthiazole (DPT) to rats. The alteration of gene expressions was analyzed by differential display. The band of decreased expression was demonstrated by differential display. By using this PCR product as a probe, we have cloned cDNA of 1482 bp from cDNA library. The sequence was identical 94.3% to rat sult1C2 cDNA. Mouse Sult1C2 mRNA was abundant in kidneys and stomachs of normal mice. The Sult1C2 gene may be involved with sulfation of proteoglycans composing the tubular basement membrane.
In order to elucidate the physiological function of Sult1C2 gene in vivo, we attempted to make Sult1C2-deficient mice. We have amplified the mouse Sul1C2 gene by PCR method and cloned for making the targeting vector. We inserted neomycin resistant gene under PGK promoter into the exon, which includes initiation codon of ATG of Sult1C2 gene. The targeting vector had two long arms of about 5 kbs for homologous recombination.
The targeting vector was introduced to mouse ES cells (R1) by electroporation and selected in neomycin containing medium. The neomycin resistant colonies were isolated and their DNAs were analyzed by Southern blot, using 5' external probe. As a result we did not isolated homologous recombinat from 500 of colonies.
We again tried to isolte the recombinant and we have isolated 2 homologous clones. This time, we tried to aggregate the ES cells with 8-cell embryos and inserted them into oviduct of pseudo-pregnant mice.
We will make chimera mice and heterogenic mice. The heterozygous mice should be crossed, and we Will get homozygous mice deficient of Sult1C2 gene.
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Research Products
(7 results)
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[Journal Article] p53 protein interacts specifically with the meiosis-specific mammalian RecA-like protein DMC1 in meiosis.2004
Author(s)
Habu, T., Wakabayashi, N., Yoshida, K., Yomogida, K., Nishimune, Y., Morita, T.
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Journal Title
Carcinogenesis 25
Pages: 889-893
Description
「研究成果報告書概要(欧文)」より
