| Budget Amount *help |
¥3,400,000 (Direct Cost: ¥3,400,000)
Fiscal Year 2002: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2001: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Research Abstract |
The mitochondrion is an organ with energy production and important function on respiratory metabolism. The acquired (aging) mutations and genetic polymorphisms of mitochondrion DNA (mtDNA) are deeply related to the disease. However, the analysis method suitable for large-scale population survey in coding region has not been established. This time, the convenient and quick method for detecting of SNPs of mtDNA by the multiplex APLP method was established.
Five SNPs (nt3010, nt4386, nt5178, nt8794, nt10398) and a 9-bp deletion (nt8272-8289) were chosen. Simultaneously, the PCR was amplified using 17 kinds of primer mix, and 6 substitutions were typed by slab-gel electrophoresis, and then stained with SYBR-Gold. In the Multiplex APLP method, simultaneously, 75-151bp PCR product was got, and it was possible to accurately judge each base substitution. B1 and C1 types were detected from all populations tested. A2, B2, B3, C2 and C4 were detected only from the Asian groups. Especially, B2 type detects considerably high-frequent from all Japanese groups. B5 and C5 were the haplotype which were almost specific in Germany. This method seemed to be a most suitable for the detection of the mtDNA haplotype, because the typing is quickly performed without requiring expensive equipments and reagents.
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