| Project/Area Number |
06807069
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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 |
Dermatology
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| Research Institution | AKITA UNIVERSITY |
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Principal Investigator |
SATO Toshiki (1995) Akita Univ.Sch.of Med, Dept.of Dermatol., Lecuturer, 医学部, 講師 (40187216)
中村 えり子 (1994) 秋田大学, 医学部, 助手 (80260454)
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| Co-Investigator(Kenkyū-buntansha) |
NIISAWA Midori Akita Univ.Sch.of Med, Dept.of Dermatol., Associate, 医学部, 助手 (00208109)
SATO Noriko Akita Univ.Sch.of Med, Dept.of Dermatol., Associate, 医学部, 助手 (60205953)
TOMITA Yasushi Akita Univ.Sch.of Med, Dept.of Dermatol., Professor, 医学部, 教授 (70108512)
窪田 卓 秋田大学, 医学部, 助手 (60250875)
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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: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1994: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | albinism / mutation / gene diagnosis / melanin / tyrosinase / 色素異常症 / 遺伝子変異 |
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| Research Abstract |
Allele-specific amplification (ASA) is a simple and non-radioactive technique for detecting known point mutations that produce genetic diseases. Although this technique is based on the specific amplification of the target allele by a polymerase chain reaction (PCR) with allele-specific primers, the specificity of the amplification may depend on various PCR conditions. To avoid non-specific amplification which leads to false-positive results in ASA,we modified both the normal and mutant allele-specific primers so that they would have one constant base mismatch, located at the penultimate 3' position. We confirmed that our modification could inhibit such unfavorable amplification by using as templates genomic DNAs of patients affected with tyrosinase-negative oculocutaneous albinism (OCA). We then analyzed new patients affected with tyrosinase-negative OCA.and based the diagnosis on both the results of a clinical examination and those of a hair bulb test using ASA with the modified allele-specific primers. The results indicated that more than 3 alleles of the tyrosinase gene with a pathological mutation existed in Japanese patients.
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