engleski

Analysis of lobel, a new gene identified by gene trap method

In order to identify and analyse new genes involved in mouse development we used a gene trap method. After insertion of the gene trap vector into an endogenous mouse gene its expression can be detected in ES cells and subsequently generated embryos. A gene expressed in developing nervous system was chosen and named lobel. Lobel is in 11.5-day embryos expressed in the neural tube and in the heart. In addition, it is expressed in the cortex of the brain of the adult mouse. The sequence of lobel was obtained by 3` and 5` RACE method. Sequence analysis revealed that the gene was novel, although it shared a part of its sequence with EST sequences. In order to characterize the way of insertion of the gene trap vector in lobel gene we used the PCR method. The insertion occured at about 1/3 length of lobel cDNA and it is located in a corresponding intron. After sequencing of the intron, we constructed the PCR-based test for homozygotes. The first homozygotes were born, and showed that the mutation of lobel caused by gene trap is not embryonic lethal.

gene trap; nervous system

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