HIV-1单循环感染

Ferdinand Roesch; Olivier Schwartz

doi:10.21769/BioProtoc.416

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Peer-reviewed

HIV-1 Single Cycle Infection

HIV-1单循环感染

FR Ferdinand Roesch

OS Olivier Schwartz email

发布: 2013年04月05日第3卷第7期 DOI: 10.21769/BioProtoc.416 浏览次数: 12681

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参见作者原研究论文

The authors used this protocol in:

Cover of PLOS Pathogens, featuring study using the protocol.

Jul 2012

Special Issue on Protocols for Organoid Research

实验方案合集

Cell Imaging - A Special Collection for Cell Bio 2023

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Abstract

The role of a viral or cellular protein on HIV-1 infection can sometimes be difficult to assess in a system where the virus is able to replicate for several cycles. Indeed, some effects are only observable at early time points, and can be masked (or on the contrary, artificially increased) after several rounds of viral replication. Therefore, to clearly know at which step of HIV-1 replication cycle one protein acts, it is important to be able to study one cycle of infection only. This protocol allows rapid and robust quantification of HIV-1 single cycle infection, and can be used to compare mutant viruses, or treatments with different drugs.

Materials and Reagents

HeLa CD4⁺ CCR5⁺LTR lacZ (also known as P4C5) cells (Verrier et al., 1997)
Dulbecco Eagle’s Modified Medium (DMEM) (Life Technologies, Gibco^®)
HIV-1 Δenv viral particles, produced by transfection of HEK-293T cells
Nonidet P40 (NP40) (Sigma-Aldrich)
No longer commercially available, but replaced by Igepal CA-630 (Sigma-Aldrich, catalog number: I8896 )
Phosphate buffer saline (PBS) (Life Technologies, Gibco^®)
Chlorophenol Red-β-D-Galactopyranoside (CPRG) (F. Hoffmann-La Roche, catalog number: 10884308001 )
Fetal bovine serum (FBS) (Thermo Fisher Scientific)
Penicillin/Streptomycin (PS) (Life Technologies, Gibco^®)