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Bisarsenical labeling of HIV-1 for real-time fluorescence microscopy.

Abstract : Imaging studies have benefited from the development of a novel technique for non-destructive labeling of proteins within living cells, based on the use of a reagent called FlAsH-EDT2, a bisarsenical derivative of fluorescein capable of binding with high affinity and specificity to a tetracysteine motif in the protein of interest. This technique has been adapted for the stable, sensitive and specific molecular tagging of HIV-1 IN enabling the tracking of incoming viral particles inside infected living cells. Here we present the experimental steps required for the efficient labeling of HIV-1 IN, namely, molecular insertion of a tetracysteine tag, production of viruses, labeling in vitro of tagged viruses, infection of target cells and visualization of particles by fluorescence microscopy.
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https://hal-pasteur.archives-ouvertes.fr/pasteur-00457792
Contributor : Mireille Gau <>
Submitted on : Thursday, February 18, 2010 - 2:43:23 PM
Last modification on : Monday, January 13, 2020 - 5:08:10 PM

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Nathalie J Arhel, Pierre Charneau. Bisarsenical labeling of HIV-1 for real-time fluorescence microscopy.. Methods in Molecular Biology, Humana Press/Springer Imprint, 2009, 485, pp.151-9. ⟨10.1007/978-1-59745-170-3_11⟩. ⟨pasteur-00457792⟩

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