| Project/Area Number |
01570163
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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 |
Pathological medical chemistry
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| Research Institution | Nagasaki University |
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Principal Investigator |
KOIKE Kichiko Nagasaki University, Sch. of Med. Associate Professor, 医学部, 助教授 (80039619)
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| Co-Investigator(Kenkyū-buntansha) |
URATA Yoshishige Nagasaki University, Sch. of Med. Instructor, 医学部, 助手 (30185087)
KOIKE Masahiko Nagasaki University, Sch. of Med. Professor, 医学部, 教授 (10039521)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 1990: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1989: ¥1,000,000 (Direct Cost: ¥1,000,000)
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| Keywords | Pyruvate dehydrogenase (PDH) / PDHalpha gene / PDHbeta gene / Transcription start point / CAAT box / TATA box / Pyruvate dehydrogenase deficiency |
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| Research Abstract |
1. Genomic clones encompassing the entire gene for human PDHalpha has been isolated from a leukocyte genomic library. The PDHalpha gene spans 17,082 base pairs and is composed of 11 exons and 10 introns. A total of seven Alu repeats was found in five introns. The entire nucleotide sequence of the gene has been determined and typical consensus promoter sequences in the 5'-flanking region were found. Primer extension analysis indicated that transcription start point is a thymine residue 124 bases upstream from ATG codon in exon 1.
2. Genomic clones encompassing the entire gene for human PDHbeta has been isolated from a leukocyte genomic library constructed into lambdaEMBL3. The clone was characterized by restriction enzyme analysis, extensive DNA sequencing and primer extension analysis. The PDHbeta gene spans 18 kilobase pairs and is composed of 10 exons and 9 introns. Two Alu repeats were found in two introns. The 5'-flanking region of the gene contains CAAT box and no TATA box. The transcription was initiated with a adenine residue 129 bases upstream from the translation initiation codon in exon 1.
3. A search for restriction fragment length polymorphisms has not led to the detection of the specific polymorphism to genome of pyruvate dehydrogenase deficiency patient. Therefore it suggested the necessity of nucleotide sequencing of all exons amplified by PCR for a search of the inherited metabolic abnormalities by deletion or displacement of nucleotide sequence.
4. We are now investigating the abnormalities of the pyruvate dehydrogenase deficiency patient's genome.
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