, Volume 9, Issue 6, pp 395-404

A New Gene Trap Construct Enriching for Insertion Events Near the 5′ End of Genes

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The gene trap approach is based on the integration of a gene trap vector into the genome. This can be done either by electroporation of a plasmid construct or by infection with a viral vector. Commonly used viral gene trap vectors have been shown to select for integrations near the 5′ end of genes. To date, no plasmid vector with a similar tendency has been reported. In this paper we describe a new plasmid vector, pKC199βgeo. This vector contained a short splice acceptor fragment from the Hoxc9 gene, a full length lacZ gene, including an ATG, and a reduced activity, mutant neomycin phosphotransferase gene as a selectable marker. This vector enriched the population of trapped genes in our gene trap screen for insertion events in the 5′ end of genes. In the two cases examined the β-galactosidase activity pattern accurately reflected the endogenous promotor activity.