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首页> 外文期刊>Applied Microbiology >Fluorescent Protein Expression as a Proxy for Bacterial Fitness in a High-Throughput Assay
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Fluorescent Protein Expression as a Proxy for Bacterial Fitness in a High-Throughput Assay

机译：荧光蛋白表达作为高通量测定中的细菌适应性的代理

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Bacterial growth is classically assessed by measuring the increases in optical density of pure cultures in shaken liquid media. Measuring growth using optical density has severe limitations when studying multistrain interactions, as it is not possible to measure the growth of individual strains within mixed cultures. Here, we demonstrated that constitutively expressed fluorescent proteins can be used to track the growth of individual strains in different liquid media. Fluorescence measurements were highly correlated with optical density measurements and cell counts. This allowed us to assess bacterial growth not only in pure cultures but also in mixed bacterial cultures and determine the impact of a competitor on a focal strain, thereby assessing relative fitness. Furthermore, we were able to track the growth of two different strains simultaneously by using fluorescent proteins with differential excitation and emission wavelengths. Bacterial densities measured by fluorescence yielded more consistent data between technical replicates than optical density measurements. Our setup employs fluorescence microplate readers that allow high throughput and replication.IMPORTANCE We expand on an important limitation of the concept of measuring bacterial growth, which is classically limited to one strain at a time. By adopting our approach, it is possible to measure the growth of several bacterial strains simultaneously with high temporal resolution and in a high-throughput manner. This is important to investigate bacterial interactions, such as competition and facilitation.
机译：通过测量摇动液体介质中纯培养物的光密度的增加来评估细菌生长。测量使用光密度的生长在研究多丙烯相互作用时具有严重的限制，因为不可能测量混合培养物中的个体菌株的生长。这里，我们证明了组成型表达的荧光蛋白可用于追踪不同液体介质中单个菌株的生长。荧光测量与光学密度测量和细胞计数高度相关。这使我们允许我们不仅在纯培养物中评估细菌生长，而且在混合细菌培养物中，并确定竞争对手对焦株的影响，从而评估相对适应性。此外，我们能够通过使用具有差分激发和发射波长的荧光蛋白来追踪两种不同菌株的生长。通过荧光测量的细菌密度在技术复制之间产生比光学密度测量值更一致的数据。我们的设置采用荧光微孔板读卡器，允许高吞吐量和复制。分析我们在测量细菌生长的概念的一个重要限制中扩展，这一次典型地限于一种菌株。通过采用我们的方法，可以以高吞吐量和高通量的方式同时测量几种细菌菌株的生长。这对于调查细菌相互作用是重要的，例如竞争和便利化。

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