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• W3004232800 endingPage "125715" @default.
• W3004232800 startingPage "125715" @default.
• W3004232800 abstract "Phaeodactylum tricornutum is an ecologically and evolutionarily relevant microalga that has developed into an important model for molecular biological studies on organisms with complex plastids. The diatom is particularly suitable for in vivo protein localization analyses via fluorescence microscopy in which the green fluorescent protein (GFP) and its derivatives are dominantly used. Whereas GFP fluorescence emission is usually measured between 500 and 520 nm in confocal microscopy, the autofluorescence of the P. tricornutum plastid is detected above 625 nm. Here we established the fluorescent protein mRuby3 as tag for efficient in vivo protein localization studies by expressing a codon-optimized gene in P. tricornutum. mRuby3 was directed to seven different subcellular localizations by means of full-length marker protein or N-/C-terminal targeting signal fusions; its emission was detected efficiently between 580 and 605 nm, being unequivocally distinguishable from the plastid autofluorescence in vivo. Moreover, mRuby3 proved to be highly suitable for co-localization experiments using confocal laser scanning microscopy in which mRuby3 fusion proteins were expressed in parallel with GFP-tagged proteins. Our results show the potential of mRuby3 for its application in studying protein targeting and localization in P. tricornutum, particularly underlining its compatibility with GFP and the plastid autofluorescence in signal detection." @default.
• W3004232800 created "2020-02-07" @default.
• W3004232800 creator A5012164892 @default.
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• W3004232800 date "2020-02-01" @default.
• W3004232800 modified "2023-09-27" @default.
• W3004232800 title "Optimized mRuby3 is a Suitable Fluorescent Protein for in vivo Co-localization Studies with GFP in the Diatom Phaeodactylum tricornutum" @default.
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• W3004232800 doi "https://doi.org/10.1016/j.protis.2020.125715" @default.
• W3004232800 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/32062589" @default.
• W3004232800 hasPublicationYear "2020" @default.
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