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Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi

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 fluoresc...

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Autores principales: Werner, Antonia, Otte, Kolja L., Stahlhut, Gertrud, Pöggeler, Stefanie
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7752937/
https://www.ncbi.nlm.nih.gov/pubmed/33349910
http://dx.doi.org/10.1186/s13568-020-01160-x
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author Werner, Antonia
Otte, Kolja L.
Stahlhut, Gertrud
Pöggeler, Stefanie
author_facet Werner, Antonia
Otte, Kolja L.
Stahlhut, Gertrud
Pöggeler, Stefanie
author_sort Werner, Antonia
collection PubMed
description 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.
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spelling pubmed-77529372020-12-28 Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi Werner, Antonia Otte, Kolja L. Stahlhut, Gertrud Pöggeler, Stefanie AMB Express Original Article 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. Springer Berlin Heidelberg 2020-12-21 /pmc/articles/PMC7752937/ /pubmed/33349910 http://dx.doi.org/10.1186/s13568-020-01160-x Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Original Article
Werner, Antonia
Otte, Kolja L.
Stahlhut, Gertrud
Pöggeler, Stefanie
Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi
title Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi
title_full Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi
title_fullStr Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi
title_full_unstemmed Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi
title_short Establishment of the monomeric yellow-green fluorescent protein mNeonGreen for life cell imaging in mycelial fungi
title_sort establishment of the monomeric yellow-green fluorescent protein mneongreen for life cell imaging in mycelial fungi
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7752937/
https://www.ncbi.nlm.nih.gov/pubmed/33349910
http://dx.doi.org/10.1186/s13568-020-01160-x
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