Loh Jacelyn M S, Proft Thomas
Department of Molecular Medicine & Pathology, School of Medical Sciences, University of Auckland, Auckland, New Zealand,
Biotechnol Lett. 2014 Apr;36(4):829-34. doi: 10.1007/s10529-013-1423-z. Epub 2013 Dec 10.
NanoLuc luciferase (Nluc) is an engineered enzyme that catalyses the substrate, furimazine, to produce light. Nluc has higher sensitivity than the commonly used bioluminescent reporter, firefly luciferase (FFluc). We have introduced Nluc into a toxin-antitoxin stabilised plasmid for the efficient labeling of group A Streptococcus. Comparison of signal strength and kinetic properties between Nluc-labeled bacteria and similarly previously-labeled FFluc bacteria, showed that the bioluminescent signal produced by Nluc-labeled bacteria is up to 15-times higher than FFluc-labeled bacteria during the logarithmic phase. However, with Nluc we were unable to differentiate between bacteria that are metabolically active and inactive because of its ATP-independence. Nluc therefore offers a more sensitive reporter but, perhaps, one more restricted for downstream applications.
纳米荧光素酶（Nluc）是一种经过改造的酶，可催化底物腔肠素，产生光。Nluc比常用的生物发光报告基因萤火虫荧光素酶（FFluc）具有更高的灵敏度。我们已将Nluc引入毒素-抗毒素稳定化质粒中，用于A群链球菌的高效标记。对Nluc标记细菌和先前类似标记的FFluc细菌之间的信号强度和动力学特性进行比较，结果表明，在对数期，Nluc标记细菌产生的生物发光信号比FFluc标记细菌高15倍。然而，由于Nluc不依赖ATP，我们无法区分代谢活跃和不活跃的细菌。因此，Nluc提供了一种更灵敏的报告基因，但可能在下游应用中受到更多限制。
