1994 Fiscal Year Final Research Report Summary
Sex determination based on detecting the difference of methylation pattern on the DXZ4 region of X chromosome
| Project/Area Number |
05670381
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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 |
Legal medicine
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| Research Institution | Niigata University |
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Principal Investigator |
MAMANOUCHI Haruo Niigata Univ.School of Medicine Professor, 医学部, 教授 (30134919)
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| Co-Investigator(Kenkyū-buntansha) |
KOMINAMI Ryo Niigata Univ.School of Medicine Professor, 医学部, 教授 (40133615)
DEWA Koji Niigata Univ.School of Medicine Assistant, 医学部, 助手 (10197832)
NAITO Emiko Niigata Univ.School of Medicine Assistant, 医学部, 助手 (80018811)
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| Project Period (FY) |
1993 – 1994
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| Keywords | DXZ4 / methylation / inactive chromosome X / PCR / female-typing / Sex identification / SRY |
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| Research Abstract |
We have developed a novel method for positively determining female sex of biological samples with polymerase chain reaction (PCR). This uses a repetitive sequence, DXZ4, on X chromosome as a typing marker. The DXZ4 region is highly methylated on the active X while it is hypomethylated on the inactive X.Hence, when DNA is digested with a methylation-sensitive enzyme, Hpa II,only the repetitive sequence on the inactive X chromosome specific to females yield small DNA fragments of approximately 100-bp. Such fragments are isolated with gel electrcphoresis and detected by PCR.In parallel, a complementary male-sex typing has been also carried out using the SRY sequence as a probe.
Testing DNAs from fresh Turner's blood and from postnortem tissues exhibited band-signals confirming the sex identification. Degraded DNAs isolated from severely decomposed specimens were also identifiable when high-molecular-weight DNA was isolated before the assay. This demonstrates the usefulness of this method in forensic identification.
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