Transgenic Research

, Volume 9, Issue 6, pp 395–404

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

  • Tim Thomas
  • Anne K. Voss
  • Kamal Chowdhury
  • Peter Gruss
Article

DOI: 10.1023/A:1026595111913

Cite this article as:
Thomas, T., Voss, A.K., Chowdhury, K. et al. Transgenic Res (2000) 9: 395. doi:10.1023/A:1026595111913

Abstract

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.

gene trap vectors 

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Tim Thomas
    • 1
  • Anne K. Voss
    • 1
  • Kamal Chowdhury
    • 1
  • Peter Gruss
    • 2
  1. 1.Department of Molecular Cell BiologyMax Planck Institute of Biophysical ChemistryGöttingenGermany
  2. 2.Department of Molecular Cell BiologyMax Planck Institute of Biophysical ChemistryGöttingenGermany