International Mammalian Genome Society

The 14th International Mouse Genome Conference (2000)

H17. Characterization of Two Mutant Mouse Lines Established by the Exchangeable Gene Trapping

K. Semba1, T. Sekimoto1,3, J. Yoshimuta1, M. Suzuki2, K. Araki1, K. Yamamura1
1Division of Developmental Genetics, Institute of Molecular Embryology and Genetics, Kumamoto University School of Medicine, 2Center for Animal Resources and Development, Kumamoto University, Kumamoto, Japan. 3Dept. of Orthopaedics Surgery, Miyazaki Medical College, Miyazaki, Japan.

Gene trap strategy using embryonic stem cell (ES) is a powerful 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 through Cre-mutated lox mediated integration. We have isolated 109 trap clones using the trap vector pU-Hachi and generated 17 mutant mouse lines. Here, we present the analysis of the two mutant mouse lines, Ayu8021 and Ayu8029.

In Ayu8021 line, the expression of the -geo gene began from E9.5 and was restricted in the notochord until E12.5. In homozygous adult mice, the -geo gene was expressed in callosal body, cardiac muscle and interstitium testis. Interestingly, homozygotes exhibited kinky tail. Abnormalities in the caudal vertebra were always observed in the 5-7th vertebrae from the end. Partial cDNA of the trapped gene was isolated by 5' rapid amplication of cDNA ends (5'-RACE). The cDNA showed homology to several EST sequences.

In Ayu 8029 line, the -geo gene was expressed ubiquitously at E10.5 embryo, and in brain, kidney and testis at adult. We obtained 3.7kb of cDNA fragment that frank 5' to the splice acceptor sequence of the trap vector by 5'-RACE. The cDNA has 100% homology to mouse pericentriolar material 1 (PCM-1).

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