1994 Fiscal Year Final Research Report Summary
GENE ANALYSIS OF FRUCTOSE-1,6-DIPHOSPHATASE DEFICIENCY
| Project/Area Number |
05670664
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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 |
Pediatrics
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| Research Institution | FUKUI MEDICAL SCHOOL |
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Principal Investigator |
KIKAWA Yoshiharu DEPARTMENT OF PEDIATRICS,FUKUI MEDICAL SCHOOL,ASSISTANT PROFESSOR, 医学部, 講師 (90143940)
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| Co-Investigator(Kenkyū-buntansha) |
MIZUNUMA Hideo DEPARTMENT OF BIOCHEMISTRY AKITA UNIVERSITY COLLEGE OF ALLIED MEDICAL SCIENCE,PR, 医療技術短期大学部, 教授 (70095389)
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| Project Period (FY) |
1993 – 1994
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| Keywords | fructose-1,6-bisphosphatase deficiency / monocytes / point mutation / expression / Insertion of G nucleotide |
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| Research Abstract |
1 : Fructose 1,6-bisphosphatase deficiency is an autosomal recessive inherited disorder of gluconeogenesis. We could isolate cDNAs encoding human tructose 1,6-bisphosphatase from normal monocytes, liver and kidney, but not from normal lymphocytes. The cDNAs contained an open reading frame coding for 338 amino acids, and their nucleotide sequences in monocytes and liver were identical.
2 : Analysis of monocytes in seven patients with fructose 1,6-bisphosphatase deficiency showed a DNA fragment with apparent normal size in two sisters but no detectable DNA fragment in the other five patients. Two discrete mutations were identified in the two sisters, products of a nonconsanguineous marriage.
3 : One is an insertion of G resulting in a reading-frame shift with truncated mutant enzyme, and the other is a transition of T to C converting Val to Ala at residue 325. The sisters were confirmed to be compound heterozygotes, inheriting one allele from their father and the other allele from their mother, both by nucleotide sequencing of the subcloned PCR products and by direct sequencing of the PCR products.
4 : These two mutant and normal clones were expressed in Esherichia coli. The mutant clone with the insertion of G showed a protein band smaller than that of normal clones, and revealed completely deficient enzyme activities. While, the other mutant clone with the transition of T to C showed a protein band with apparently normal size, and retained normal FBPase activitiy. This report appears to be a first molecular analysis on genetic defects in patients with FBPase deficiency.
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