| Project/Area Number |
11671133
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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 |
Metabolomics
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| Research Institution | Kyoto Prefectural University of Medicine |
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Principal Investigator |
NOSAKA Kazuto Kyoto Pref. Univ. Med., Dept. Chem., Associate Prof., 医学部, 助教授 (10228314)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥2,800,000 (Direct Cost: ¥2,800,000)
Fiscal Year 2000: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1999: ¥1,700,000 (Direct Cost: ¥1,700,000)
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| Keywords | thiamin / thiamin-responsive megaloblastic anemia / thiamin pyrophosphokinase / thiamin transport protein / thiamin pyrophosphate |
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| Research Abstract |
Thiamin-responsive megaloblastic anemia (TRMA ; OMIM 249270) is an autosomal recessive disorder defined by the occurrence of megaloblastic anemia, diabetes mellitus, and sensorineural deafness, responding to thiamin treatment. It is highly possible that thiamin pyrophosphokinase (TPK) or thiamin transport system is a candidate for the aberrant protein.
Recently, the TRMA gene was identified by positional cloning. Mutations in the patients, including a Japanese family, were found in a thiamin transporter gene (THTR-1). On the other hand, we isolated a mouse thiamin pyrophosphokinase cDNA clone (mTPK1) using a combination of mouse expressed sequence tag database analysis, two-step polymerase chain reaction procedure and functional complementation screening with Saccharomyces cerevisiae thiamin pyrophosphokinase-deficient mutant (thi80). The human cDNA (hTPK1) was subsequently isolated by colony hybridization using the mTPK1 clone. The predicted protein encoded by the hTPK1 ORF contained 243 amino acid residues whose sequence showed 89% identity with that of mTPK1 product. The hTPK1 mRNA was widely expressed in various human tissues at very low level, and the mRNA content in cultured TRMA fibroblasts was unaffected by the thiamin concentration of the medium. In addition, we assigned the chromosomal localization of the hTPK1 gene to 7q34 by FISH procedure. It is expected that basic scientific research using the hTPK1 cDNA will enhance the understanding of human thiamin metabolism.
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