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红色荧光蛋白变体,具有双峰荧光发射。

Red Fluorescent Protein Variant with a Dual-Peak Emission of Fluorescence.

机构信息

Departamento de Biología, Universidad del Valle (UV), Cali, Valle del Cauca, Colombia.

Laboratório de Biologia Molecular, Instituto de Ciências Biológicas, Universidade Federal do Rio Grande (FURG), Rio Grande, RS, Brazil.

出版信息

Mar Biotechnol (NY). 2023 Dec;25(6):1099-1109. doi: 10.1007/s10126-023-10262-z. Epub 2023 Oct 21.

Abstract

The marine environment is a rich reservoir of diverse biological entities, many of which possess unique properties that are of immense value to biotechnological applications. One such example is the red fluorescent protein derived from the coral Discosoma sp. This protein, encoded by the DsRed gene, has been the subject of extensive research due to its potential applications in various fields. In the study, a variant of the red fluorescent protein was generated through random mutagenesis using the DsRed2 gene as a template. The process employed error-prone PCR (epPCR) to introduce random mutations, leading to the isolation of twelve gene variants. Among these, one variant stood out due to its unique spectral properties, exhibiting dual fluorescence emission at both 480 nm (green) and 550 nm (red). This novel variant was expressed in both Escherichia coli and zebrafish (Danio rerio) muscle, confirming the dual fluorescence emission in both model systems. One of the immediate applications of this novel protein variant is in ornamental aquaculture. The dual fluorescence can serve as a unique marker or trait, enhancing the aesthetic appeal of aquatic species in ornamental settings.

摘要

海洋环境是一个拥有丰富多样生物实体的宝库,其中许多生物都具有独特的性质,对生物技术应用具有巨大的价值。例如,来自珊瑚 Discosoma sp. 的红色荧光蛋白就是一个很好的例子。这种蛋白质由 DsRed 基因编码,由于其在各个领域的潜在应用,已经成为广泛研究的对象。在这项研究中,通过使用 DsRed2 基因作为模板的易错 PCR(epPCR),产生了一种红色荧光蛋白的变体。该过程引入了随机突变,导致分离出了 12 种基因变体。其中一个变体由于其独特的光谱特性而脱颖而出,它在 480nm(绿色)和 550nm(红色)处都能发出双荧光。这种新型变体在大肠杆菌和斑马鱼(Danio rerio)肌肉中都有表达,在这两个模型系统中都证实了双荧光的发射。这种新型蛋白质变体的一个直接应用是在观赏水产养殖中。这种双荧光可以作为一种独特的标记或特征,增强观赏鱼类在观赏环境中的美学吸引力。

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