Abstract
Nef is a multifunctional protein encoded by all primate lentiviruses that modulates cell surface expression of a variety of cellular receptors and increases the infectivity of progeny virons. Here, we describe the use of bicistronic HIV-1 constructs that coexpress Nef and fluorescent proteins via an internal ribosome entry site to quantify Nef-mediated receptor modulation in virally infected cells. We also report how such proviral constructs and indicator cell lines can be used to quantify the effect of Nef on virion infectivity.
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Acknowledgments
This work was supported by the Deutsche Forschungsgemeinschaft. pBR-HIV-1 NL4-3 nef IRES eGFP reporter constructs, P4-R5 MAGI and TZM-bl cells can be obtained from the AIDS reagent program (http://www.aidsreagent.org/).
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Heigele, A., Sauter, D., Münch, J., Kirchhoff, F. (2014). HIV-1 Accessory Proteins: Nef. In: Vicenzi, E., Poli, G. (eds) Human Retroviruses. Methods in Molecular Biology, vol 1087. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-670-2_10
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DOI: https://doi.org/10.1007/978-1-62703-670-2_10
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-669-6
Online ISBN: 978-1-62703-670-2
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