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Measurement of hygromycin B phosphotransferase activity in crude mammalian cell extracts by a simple dot-blot assay

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  • M S Sørensen, Denmark
  • M Duch, Denmark
  • K Paludan, Denmark
  • Poul Jørgensen, Denmark
  • F S Pedersen, Denmark
  • Interdisciplinary Nanoscience Center
  • Department of Science Studies
  • Department of Molecular Biology
Hygromycin B (Hy) resistance, encoded by the prokaryotic gene hph, is commonly used as a dominant selectable marker for gene transfer experiments in mammalian cells. We describe a simple, quantitative dot-blot assay for measuring the activity in crude mammalian cell extracts of Hy phosphotransferase, the product of the hph gene. The assay shows no cross interference with substrates for neomycin phosphotransferase II, the product of the commonly used marker gene neo; hph and neo may thus be useful as a set of two non-interfering selectable marker and reporter genes for gene transfer experiments in mammalian cells.
Original languageEnglish
Pages (from-to)257-60
Number of pages3
Publication statusPublished - 1992

    Research areas

  • Animals, Cell Extracts, Cell Line, Genetic Markers, Kanamycin Kinase, Phosphotransferases, Phosphotransferases (Alcohol Group Acceptor), Substrate Specificity

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