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Company: Sigma-Aldrich

Catalog#: P9861

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Analysis of Cleavage Activity of Dengue Virus Protease by Co-transfections

Lekha Gandhi and Musturi Venkataramana

Published online: 2024-03-05

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HIV-1 Fusion Assay

Author:

Marielle Cavrois, Jason Neidleman and Warner C. Greene,

Date:

2014-08-20

[Abstract] The HIV-1 fusion assay measures all steps in the HIV-1 life cycle up to and including viral fusion. It relies on the incorporation of a β-lactamase–Vpr (BlaM-Vpr) protein chimera into the virion and the subsequent transfer of this chimera into the target cell by fusion (Figure 1). The transfer is monitored by the enzymatic cleavage of CCF2, a fluorescent dye substrate of β-lactamase, loaded into the target cells. Cleavage of the β-lactam ring in CCF2 by β-lactamase changes the fluorescence emission spectrum of the dye from green (520 nm) to blue (447 nm). This change reflects virion fusion and can be detected by flow cytometry (Figure 2). [摘要] HIV-1融合测定测量HIV-1生命周期中直至并包括病毒融合的所有步骤。它依赖于将β-内酰胺酶-Vpr（BlaM-Vpr）蛋白嵌合体并入病毒体中，并且随后通过融合将该嵌合体转移到靶细胞中（图1）。通过CCF2（一种β-内酰胺酶的荧光染料底物）的酶裂解来监测转移，将其装载到靶细胞中。 β-内酰胺酶在CCF2中的β-内酰胺环的切割将染料的荧光发射光谱从绿色（520nm）改变为蓝色（447nm）。这种变化反映了病毒体融合，并且可以通过流式细胞术检测（图2）。

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