| Project/Area Number |
14580796
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Laboratory animal science
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| Research Institution | Shinshu University |
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Principal Investigator |
MORI Masayuki Shinshu University, School of Medicine, Associate professor, 医学部, 助教授 (60273190)
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| Co-Investigator(Kenkyū-buntansha) |
HIGUCHI Keiichi Shinshu University, School of medicine, Professor, 医学部, 教授 (20173156)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥2,300,000 (Direct Cost: ¥2,300,000)
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| Keywords | retrotransposon / rat / L1 / mutation / Chediak-Higashi / Lyst |
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| Research Abstract |
We have obtained the following data on L1b sequence found in the rat lysosomal trafficking regulator (Lyst) gene.
1.Molecular genetic analysis of the Lyst gene of -ACI-beige rats, which is a model of human. Chediak-Higashi syndrome revealed deletion of exons 28,29,and 30 of the gene owing to recombination between L1 elements in the gene of the mutant rats. Nucleotide sequence analysis of one of the L1 elements (tentatively designated L1b) revealed that it had two intact at open reading frames (ORF1 and ORF2), a promotor-like sequence with Sp1 box in its 5' upstream region, a poly A sequence, in its 3' downstream region, and short direct repeats on up-and downstream region, which are hallmarks of active retrotransposons.
2.The rat Lyst gene was mapped on the telomeric region of chromosome 17 by genetic linkage analysis utilizing (DA/Ham-Lyst^<bg> x BN)x DA/Ham-Lyst^<bg> baclcross rats.
3.An allele-specific genotyping method for the Lyst gene, which enabled discrimination of the mutant Lyst^<bg>, Lyst^<bg-Kyo> alleles, and the normal Lyst allele, was developed. It was also demonstrated that the deletion break points in L1 elements were different between the Lyst^<bg> and Lyst^<bg-Kyo> alleles.
4.Serial analysis gene expression (SAGE) analysis revealed that the transcription level of L1 in the mouse testis was very low.
5.Retrotransposon activity was demonstrated in the assay utilizing HeLa cell culture assay.
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