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Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi
AMB Express ( IF 3.5 ) Pub Date : 2020-12-21 , DOI: 10.1186/s13568-020-01160-x
Antonia Werner 1 , Kolja L Otte 1 , Gertrud Stahlhut 1 , Stefanie Pöggeler 1
Affiliation  

The engineered monomeric version of the lancelet Branchiostoma lanceolatum fluorescent protein, mNeonGreen (mNG), has several positive characteristics, such as a very bright fluorescence, high photostability and fast maturation. These features make it a good candidate for the utilization as fluorescent tool for cell biology and biochemical applications in filamentous fungi. We report the generation of plasmids for the expression of the heterologous mNG gene under the control of an inducible and a constitutive promoter in the filamentous ascomycete Sordaria macrospora and display a stable expression of mNG in the cytoplasm. To demonstrate its usefulness for labeling of organelles, the peroxisomal targeting sequence serine-lysine-leucine (SKL) was fused to mNG. Expression of this tagged version led to protein import of mNG into peroxisomes and their bright fluorescence in life cell imaging.



中文翻译:


用于菌丝真菌生命细胞成像的单体黄绿色荧光蛋白 mNeonGreen 的建立



文昌鱼文昌鱼荧光蛋白 mNeonGreen (mNG) 的工程单体版本具有多种积极特性,例如非常明亮的荧光、高光稳定性和快速成熟。这些特征使其成为丝状真菌细胞生物学和生化应用中荧光工具的良好候选者。我们报道了在丝状子囊菌Sordaria macrospora中诱导型和组成型启动子控制下用于表达异源 mNG 基因的质粒的产生,并在细胞质中表现出 mNG 的稳定表达。为了证明其在标记细胞器方面的有用性,将过氧化物酶体靶向序列丝氨酸-赖氨酸-亮氨酸 (SKL) 与 mNG 融合。该标记版本的表达导致 mNG 蛋白导入过氧化物酶体,并在生命细胞成像中产生明亮的荧光。

更新日期:2020-12-22
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