engleski

Gene trap modification of Nol1 (nucleolar protein 1) – a tool for analysis of Nol1 expression and function

Gene trap method enables investigation of gene expression and function using random mutagenesis and molecular tagging. In order to characterize genes involved in mouse embryo development, embryonic stem (ES) cells were modified by a nonhomologous DNA vector containing a splice acceptor and fused promoterless lacZ and neoR genes. ES cells were selected and corresponding mice were obtained. One of these was a mouse line in which strong intracellular expression of the modified gene was localized in the nucleolus. The gene was identified as nucleolar protein 1 (Nol1). Nol1 is suggested to be involved in ribosome biogenesis, and as a proliferation marker has a prognostic value in human cancer. The detailed molecular analysis of the insertion site revealed that tandem repeats of gene trap vector were located within 15th exon of Nol1 gene. Nol1 expression pattern was determined by whole mount histochemical staining by beta-galactosidase substrate X-gal. Expression was found all over the embryo, but not in every cell in all investigated developmental stages. In order to get insight in the function of Nol1, intercross of heterozygous mice carrying gene trap insertion was performed. Homozygous mice were not present among newborn animals, therefore we propose that Nol1 is required for normal nucleolar function during embryo development.

Nol1; gene trap

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