1990 Fiscal Year Final Research Report Summary
Molecular Genetic Study on Adrenoleukodystrophy
| Project/Area Number |
01440042
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| Research Category |
Grant-in-Aid for General Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
Neurology
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| Research Institution | Niigata University |
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Principal Investigator |
MIYATAKE Tadashi Niigata University Brain Research Institute Professor Department of Neurology, 脳研究所, 教授 (50048998)
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| Co-Investigator(Kenkyū-buntansha) |
OHNO Tsukasa Brain Research Institute Assistant Department of Neurology, 脳研究所, 助手 (50213814)
TSUJI Shoji Niigata University Hospital Assistant Department of Neurology, 医学部附属病院, 助手 (70150612)
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| Project Period (FY) |
1989 – 1990
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| Keywords | Adrenoleukodystrophy / Reverse Genetics / Linking Clone / CpG Island / Transcribed Sequences / Color Pigment Gene / Human X Chromosome / Very Long Chain Fatty Acid |
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| Research Abstract |
Adrenoleukodystrophy (ALD) is an X-linked recessive neurologic disease. Previous linkage studies have indicated that the ALD gene is tightly linked to Xq28 markers including G6PD and St14. To identify the ALD gene by "reverse genetics" approach, we have developed the following strategies. First, we have constructed a cosmid genomic library from a somatic cell hybrid, X3000-11, carrying only a part of human X chromosome (Xq24-28) as the human chromosome. We have isolated 1,778 human Xq24-28 cosmid clones and prepared DNA from all the cosmid clones.
We have identified 400 NotI linking clones and 350 clones carrying CpG islands, which should facilitate the identification of all house keeping genes as well as some tissue specific genes.
To efficiently identify messenger RNA transcribed from Xq24-28, we have constructed a hncDNA (heterogenous nuclear RNA-derived cDNA) library from the X3000-11 utilizing primers specific for spliceーsites. We have identified 534 hncDNA clones and confirmed that all hncDNA clones characterized are derived from Xq24-28 region.
Previous findings that color blindness is frequently found in ALD, suggests that the ALD gene might be very close to the color pigment gene. We have developed ALD cell bank and analyzed genomic DNA of 13 ALD patients. We have found a total deletion of green pigment gene in an ALD patient, which has not been described. If the ALD gene and the green pigment gene are simultaneously deleted, the case might facilitate the identification of the ALD gene.
To develop new therapeutic strategy to lower very long chain fatty acids, we have developed a new assay for evaluation of metabolic inhibitors. We have found that monoーunsaturated fatty acids with carbon numbers 20 and 22 lower the very long chain fatty acids than oleic acid.
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