| Project/Area Number |
17590583
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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 |
Legal medicine
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| Research Institution | Iwate Medical University |
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Principal Investigator |
NAKAYASHIKI Nori Iwate Medical University, School of Medicine, Department of Legal Medicine, Assistant Professor, 医学部, 講師 (10146029)
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| Co-Investigator(Kenkyū-buntansha) |
AOKI Yasuhiro Iwate Medical University, School of Medicine, Department of Legal Medicine, Professor, 医学部, 教授 (90202481)
HASHIYADA Masaki Tohoku University, Graduate School of Medicine, Division of forensic Medicine, Department of Public Health and Forensic Medicine, Technical Expert Stuff, 大学院医学研究科, 技術専門職員 (40374938)
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| Project Period (FY) |
2005 – 2006
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| Project Status |
Completed (Fiscal Year 2006)
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| Budget Amount *help |
¥2,500,000 (Direct Cost: ¥2,500,000)
Fiscal Year 2006: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2005: ¥2,000,000 (Direct Cost: ¥2,000,000)
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| Keywords | Forensic Medicine / Genomic Imprinting / Parent-of-Origin / DMR / DNA polymorphism / ゲノム・インプリンティング / 親判別 / メチル化 / クローニング |
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| Research Abstract |
1.In-the region 5'upstream-of the H19 gene, eight haplotypes composed of 14 SNPs were discovered to be applicable to the detection method of differentially methylated parental allele (DMPA). Application of DMPA detection method to new candidate SNPs in eight imprinted genes was failed.
2. Allele distributions of SNPs in four imprinted genes (H19, SNRPN, PEG3 and MEST) were examined. As a result, haplotype polymorphism of H19FR and SNPs in SNRPN and PEG3 revealed higher values of expected hetelozygosity. Multiplex PCR and mini sequencing method was used to simultaneous genotyping of above three loci plus HYMAI SNP. All seven SNPs in the four loci were successfully typed not only in native DNA but also in enzyme-digested DNA for discrimination of multilocus DMPA.
3. Methylated situation of CpGs around several SNPs in five imprinted genes was analyzed by using bisulfite-sequencing method at clonal level. In general, either paternal or maternal allele was expectedly classified into highly methylated or hypo-methylated situation according to each expression manner of the gene. However, some CpGs in the tested region showed opposite methylation status in several clones. Inconsistency of CpG methylation was considerably observed in PEG3 clones.
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