Journal ArticleDOI
Specific Covalent Labeling of Recombinant Protein Molecules Inside Live Cells
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TLDR
This system provides a recipe for slightly modifying a target protein so that it can be singled out from the many other proteins inside live cells and fluorescently stained by small nonfluorescent dye molecules added from outside the cells.Abstract:
Recombinant proteins containing four cysteines at the i , i + 1, i + 4, and i + 5 positions of an α helix were fluorescently labeled in living cells by extracellular administration of 4′,5′-bis(1,3,2-dithioarsolan-2-yl)fluorescein. This designed small ligand is membrane-permeant and nonfluorescent until it binds with high affinity and specificity to the tetracysteine domain. Such in situ labeling adds much less mass than does green fluorescent protein and offers greater versatility in attachment sites as well as potential spectroscopic and chemical properties. This system provides a recipe for slightly modifying a target protein so that it can be singled out from the many other proteins inside live cells and fluorescently stained by small nonfluorescent dye molecules added from outside the cells.read more
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Patent
Methods and compositions for synthesis of nucleic acid molecules using multiple recognition sites
Jonathan D. Chesnut,John Carrino,Louis Leong,Knut R. Madden,Martin A. Gleeson,James Fan,Michael A. Brasch,David Cheo,James L. Hartley,Devon R N Byrd,Gary F. Temple +10 more
TL;DR: In this article, the authors present compositions and methods for recombinational cloning of two or more different nucleic acid molecules, which may be fused together while in other embodiments the molecules are inserted into distinct sites in a vector.
Journal ArticleDOI
Photophysics of a water-soluble rylene dye: Comparison with other fluorescent molecules for biological applications
Anca Margineanu,Johan Hofkens,Mircea Cotlet,Satoshi Habuchi,Alina Stefan,Jianqiang Qu,Christopher Kohl,Klaus Müllen,Jo Vercammen,Yves Engelborghs,Thomas Gensch,Frans C. De Schryver +11 more
TL;DR: The photophysical properties of a new water-soluble fluorescent dye based on the perylene diimide chromophore were investigated at the ensemble and single molecule level as discussed by the authors, where the fluorophore has an absorption maximum at 567 nm and a high quantum yield of fluorescence (0.6).
Patent
Expanding the eukaryotic genetic code
TL;DR: In this paper, the authors provide compositions and methods for producing translational components that expand the number of genetically encoded amino acids in eukaryotic cells, such as orthogonal tRNAs, orthoglobal aminoacyl-tRNA synthetases and unnatural amino acids.
Journal ArticleDOI
Highly activatable and environment-insensitive optical highlighters for selective spatiotemporal imaging of target proteins
Tomonori Kobayashi,Toru Komatsu,Mako Kamiya,Claudia Guimarães Camargo Campos,Claudia Guimarães Camargo Campos,Marcos González-Gaitán,Takuya Terai,Kenjiro Hanaoka,Tetsuo Nagano,Yasuteru Urano +9 more
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Journal ArticleDOI
Selective N-terminal functionalization of native peptides and proteins.
TL;DR: A highly site-selective modification of peptides/proteins with aldehydes or carbohydrates under mild conditions was achieved.
References
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Journal ArticleDOI
The green fluorescent protein
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Journal ArticleDOI
Rapid colorimetric assay for cell growth and survival. Modifications to the tetrazolium dye procedure giving improved sensitivity and reliability.
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Journal ArticleDOI
Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin
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TL;DR: New fluorescent indicators for Ca2+ that are genetically encoded without cofactors and are targetable to specific intracellular locations are constructed and dubbed ‘cameleons’.
Journal ArticleDOI
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TL;DR: The green fluorescent protein (GFP) from the Pacific Northwest jellyfish Aequorea victoria has generated intense interest as a marker for gene expression and localization of gene products.
Journal ArticleDOI
Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer
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TL;DR: The results demonstrate that the production of more and better GFP variants is possible and worthwhile, and facilitates multicolor imaging of differential gene expression, protein localization or cell fate.
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