The Norwegian College of Fishery Science, Faculty of Biosciences, Fisheries and Economics, UiT The Arctic University of Norway, N-9037 Tromsø, Norway.
Biosensors (Basel). 2023 Feb 7;13(2):232. doi: 10.3390/bios13020232.
Reporter genes are important tools in many biological disciplines. The discovery of novel reporter genes is relatively rare. However, known reporter genes are constantly applied to novel applications. This study reports the performance of the bilirubin-dependent fluorescent protein UnaG from the Japanese eel in live cells in response to the disruption of outer membrane (OM) integrity at low bilirubin (BR) concentrations. Using the wild-type strain MC4100, its isogenic OM-deficient mutant strain NR698, and different OM-active compounds, we show that BR uptake and UnaG fluorescence depend on a leaky OM at concentrations of 10 µM BR and below, while fluorescence is mostly OM integrity-independent at concentrations above 50 µM BR. We suggest that these properties of the UnaG-BR couple might be applied as a biosensor as an alternative to the OM integrity assays currently in use.
报告基因是许多生物学领域的重要工具。新的报告基因的发现相对较少。然而,已知的报告基因不断被应用于新的应用。本研究报告了在低胆红素(BR)浓度下,日本鳗鲡胆红素依赖性荧光蛋白 UnaG 在活细胞中外膜(OM)完整性受到破坏时的性能。使用野生型菌株 MC4100、其同源 OM 缺陷突变体菌株 NR698 和不同的 OM 活性化合物,我们表明 BR 的摄取和 UnaG 荧光依赖于 10 µM BR 及以下浓度时的漏 OM,而在 50 µM BR 以上浓度时荧光主要与 OM 完整性无关。我们认为,这种 UnaG-BR 偶联物的特性可以作为生物传感器应用,作为目前使用的 OM 完整性测定的替代方法。