| Co-Investigator(Kenkyū-buntansha) |
ORII Tadao Gifu University, Dept.of Pediatrics, Professor, 医学部, 教授 (20045339)
FUKAO Toshiyuki Gifu University, Dept.of Pediatrics, Research Associate, 医学部, 助手 (70260578)
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| Research Abstract |
Beta-ketothiolase deficiency (3KTD) is an autosomal recessive metabolic disease of organic acids involving isoleucine catabolic intermediates and ketone bodies, caused by a deficiency of mitochondrial aceroacetyl-CoA thiolase (MAAT) . In this research project, we studied on the molecular basis of 3KTD,phenotype/genotype correlation, and application of molecular techniques for the diagnosis of this disease. The following results were obtained :
1) Several novel mutations that would give clues for splicing mechanisms of the RNA were identified in 3KTD : eg. a) an exonic mutation at-13 that is a nonsense mutation, causing simultaneously an exon skipping ; b) two mutations, N158D and A301P,from which translated peptides had slower electrophoretic mobilities compared with that of the normal MAAT peptide ; c) a novel 68-bp deletion involving 3' splice site of intron 7, causing exon 8 skipping.
2) We identified 19 types of gene mutations in 14 families with 3KTD,and concluded that the gene mutations as well as clinical findings were highly heterogenous among 3KTD patients. Furthermore, it was seemed that there is no phenotype/genotype correlation.
3) We developped a method for heterozygote detection using immunoblot analysis, and performed a prenatal diagnosis of this disease by PCR/MDE gel electrophoresis for the first time, as clinical applications of the molecular studies of this disease.
4) We prepared an antibody against human cytosolic acetoacety-CoA thiolase (CT) and cloned human CT cDNA.These should certainly contribute the molecular study of CT deficiency, which is another ketone body metabolic disease in the related field of 3KTD.
This study made clear the causes and mechanisms of 3KTD at the molecular level, and would enable clinical application for the diagnosis of patients with 3KTD and related ketone body disorders as well as their heterozygotes.
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