| Project/Area Number |
06670794
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Pediatrics
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| Research Institution | Gene Research Center, Tottori University |
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Principal Investigator |
NANBA Eiji Tottori University, Gene Research Center, Associated Professor, 遺伝子実験施設, 助教授 (40237631)
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| Co-Investigator(Kenkyū-buntansha) |
AKABOSHI Shinjiro Tottori University, Faculty of Medicine, Institute of Neurological Sciences, Div, 医学部, 助手 (90231810)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1995: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | myotonic dystrophy / triplet repeat disease / competitive PCR / gene expression / protein expression / 遺伝子 / RT-PCR法 |
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| Research Abstract |
1, An method of the prenatal diagnosis for congenital myotonic dystrophy.
We established the non-radioisotope PCR and Southern blot method to analyze CTG repeat size abnormality of the myotonin/protein kinase gene for the diagnosis of the patient. The CTG repeat size and flanking markers of this gene were analyzed for the prenatal diagnosis. Principally, these methods were also used for the analysis the gene abnormality of fragile X syndrome and Huntington disease in which the mechanism of the gene abnormality are similar.
2, Analysis of the myotonin/protein kinase gene expression in the lymphoblasts of the disease.
We analyzed the myotonin/protein kinase mRNA in lymphoblasts by a PCR technology. A competitive PCR technology was developed to compare the amount of mRNA between the normal and the congenital myotonic dystrophy samples. The expression level of the myotonin/protein kinase gene was not different in the normal and the disease samples. The mRNA amount of the myotonin/protein kinase gene was very low in the lymphoblasts. We will try to use the muscle for this study.
3, Expression of the myotonin/protein kinase protein in the E.coli.
We tried to make the myotonin/protein kinase protein by a E.coli expression system. The full length myotonin/protein kinase cDNAs were prepared from the human and mouse, and these cDNAs were subcloned into pRSET vectors in the system of E.coli expression. The myotonin/protein kinase protein was not detected in this system clearly. We will try to use the partial fragment of the gene or use other expression system.
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