| Project/Area Number |
07670839
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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 |
Pediatrics
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| Research Institution | CHIBA UNIVERSITY |
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Principal Investigator |
YAMAMOTO Shigenori CHIBA UNIVERSITY SCHOOL OF MEDICINE,DEPARTMENT OF PEDIATRICS,ASSISTANT, 医学部・附属病院, 助手 (30241970)
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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 |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 1996: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1995: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | CARNITINE PALMITOYLTRANSFERASE II / GENE ANALYSIS / MISSENSE MUTATION / TRANSFECTION / GENE POLYMORPHISM / 遺伝子導入 |
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| Research Abstract |
Carnitine palmitoyltransferase (CPT) II deficiency results in two different clinical forms, one with "systemic" and the other with "muscular" symptoms. We studied the molecular basis of the "systemic" form in two Japanese siblings. Their CPT II activity in lymphoblasts was reduced to 3% of the level observed in normal controls. cDNA analysis showed that the proband was a compound heterozygote. One allele carried a new mutation, G621A (E174K). The other carried three single base substitutions ; a new mutation, T1249A (F383Y) and two previously reported polymorphisms, T1156G (F352C) and G1203A (V368I). The brother had the same four substitutions. Neither of the two new mutations in this study were detected in the 60 alleles of 30 Japanese control subjects. Secondary structure prediction analysis of the mutated CPT II protein was quite different from the normal protein. To study the effects of these putative disease causing mutations on the activity of human CPT II,the substitutions were introduced into the CPT II cDNA,and three CPT II cDNA were transiently expressed in COS-7 cells using pCAGGS vector. The increases of the CPT II activity in cells after the transfections with the E174K and F383Y mutants were both 30% of that with the wild type. We concluded that E174K and F383Y were disease causing mutations.
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