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アクティブボード・２００３年３月
研究発表を行った学会；16TH INTERNATIONAL MOUSE GENOME CONFERENCE
２００２年１１月１７日～２１日（San Antonio, Texas）
タイトル；The novel gene expressed in notochord is isolated by the exchangeable gene trap method.
発表者；仙波 圭 氏
（熊本大学・発生医学研究センター・臓器形成分野）
Abstract；
Gene trap strategy using embryonic stem cell (ES) is useful method for both identification of genes and subsequent establishment of mutant lines. We have established exchangeable gene trap system, in which the reporter gene of the integrated trap vector can be replaced into any gene of interest though Cre-mutated lox mediated integration.
Here, we present the analysis of the mutant mouse line, Ayu8021 which was established by the exchangable gene trap method. In Ayu8021 line, the expression of the beta-geo gene was restricted in the notochord at embryogenesis. In mutant adult mice, the beta-geo gene was expressed strongly in nucleus pulposus of caudal intervertebral disc. Interestingly, homozygotes exhibited kinked tail at the tip. The cDNA isolated by RACE showed homology to several EST sequences. We obtained the most part of the cDNA by RT-PCR. The sequences showed an ORF of 4056 nucleotides and were capable of encoding a 1352 amino acid polypeptide with a predicted molecular weight of 147kDa. This gene was constructed from at least 22exons and was coded on 0.5Mb genomic region. Analysis of the mutated genomic region revealed that the trap vector insertion occured in an intron of the gene located between the 15th exon and 16th exon. Purified anti-the trapped gene antibodies recognized a 150kDa protein present in extracts of the trapped gene expression vector transfected BMT10. In histological analysis, the trapped gene expressed at cytoplasm in nucleus pulposus of caudal intervertebral disc.
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