The β-Lactamase Assay: Harnessing a FRET Biosensor to Analyse Viral Fusion Mechanisms.
The β-Lactamase Assay: Harnessing a FRET Biosensor to Analyse Viral Fusion Mechanisms.
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DOI：
10.3390/s16070950
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发表时间：
2016-06-23
期刊：
影响因子：
--
通讯作者：
Padilla-Parra S
中科院分区：
文献类型：
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作者：
Jones DM;Padilla-Parra S
β-内酰胺酶（BlaM）测定于1998年首次发现，并被证明是一个强大的Förster共振能量转移（FRET）为基础的报告系统，与一系列常用的细胞系兼容。今天，BlaM检测作为一种试剂盒在商业上可用，并且可以方便和廉价地用于需要基于荧光的读数的一系列实验程序。BlaM试验的一个常见应用是测量病毒融合——病毒颗粒内的遗传物质在成功进入细胞质后释放到细胞质中的时刻。该系统的灵活性不仅允许评估总的融合水平，而且还允许评估融合动力学。然而，在科学文献中，关于将该检测应用于病毒融合分析的方法存在显著差异，使得结果之间的比较变得困难。在这篇综述中，我们提请注意这些方法的差异，并检查每种方法的优点和缺点。还讨论了使病毒与基于blam的分析兼容的成功策略。
The β-lactamase (BlaM) assay was first revealed in 1998 and was demonstrated to be a robust Förster resonance energy transfer (FRET)-based reporter system that was compatible with a range of commonly-used cell lines. Today, the BlaM assay is available commercially as a kit and can be utilised readily and inexpensively for an array of experimental procedures that require a fluorescence-based readout. One frequent application of the BlaM assay is the measurement of viral fusion—the moment at which the genetic material harboured within virus particles is released into the cytosol following successful entry. The flexibility of the system permits evaluation of not only total fusion levels, but also the kinetics of fusion. However, significant variation exists in the scientific literature regarding the methodology by which the assay is applied to viral fusion analysis, making comparison between results difficult. In this review we draw attention to the disparity of these methodologies and examine the advantages and disadvantages of each approach. Successful strategies shown to render viruses compatible with BlaM-based analyses are also discussed.