Engineering green fluorescent protein for improved brightness, longer wavelengths and fluorescence resonance energy transfer
Roger Heim,Roger Y. Tsien +1 more
TLDR
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.About:
This article is published in Current Biology.The article was published on 1996-02-01 and is currently open access. It has received 1559 citations till now. The article focuses on the topics: Green fluorescent protein & Bimolecular fluorescence complementation.read more
Citations
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The green fluorescent protein
TL;DR: In just three years, the green fluorescent protein from the jellyfish Aequorea victoria has vaulted from obscurity to become one of the most widely studied and exploited proteins in biochemistry and cell biology.
Journal ArticleDOI
Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae
Mark S. Longtine,Amos Mckenzie,Douglas J. Demarini,Nirav Shah,Achim Wach,Arndt Brachat,Peter Philippsen,John R. Pringle +7 more
TL;DR: A new set of plasmids that serve as templates for the PCR synthesis of fragments that allow a variety of gene modifications that should further facilitate the rapid analysis of gene function in S. cerevisiae.
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Fluorescent indicators for Ca2+based on green fluorescent proteins and calmodulin
Atsushi Miyawaki,Juan Llopis,Roger Heim,J. Michael McCaffery,Joseph A. Adams,Mitsuhiko Ikura,Mitsuhiko Ikura,Roger Y. Tsien +7 more
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’.
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Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus
Shankar Srinivas,Tomoko Watanabe,Chyuan-Sheng Lin,Chris M William,Yasuto Tanabe,Thomas M. Jessell,Frank Costantini +6 more
TL;DR: In contrast to existing lacZ reporter lines, where lacZ expression cannot easily be detected in living tissue, the EYFP and ECFP reporter strains are useful for monitoring the expression of Cre and tracing the lineage of these cells and their descendants in cultured embryos or organs.
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Visualizing secretion and synaptic transmission with pH-sensitive green fluorescent proteins
TL;DR: PHluorins are developed pH-sensitive mutants of green fluorescent protein by structure-directed combinatorial mutagenesis, with the aim of exploiting the acidic pH inside secretory vesicles, to monitor vesicle exocytosis and recycling.
References
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Green fluorescent protein as a marker for gene expression
TL;DR: A complementary DNA for the Aequorea victoria green fluorescent protein produces a fluorescent product when expressed in prokaryotic or eukaryotic cells, which can be used to monitor gene expression and protein localization in living organisms.
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Primary structure of the Aequorea victoria green-fluorescent protein.
Douglas Prasher,Virginia K. Eckenrode,William W. Ward,Frank G. Prendergast,Milton J. Cormier +4 more
TL;DR: The cloning and sequencing of both cDNA and genomic clones of GFP from the cnidarian, Aequorea victoria, show three different restriction enzyme patterns which suggests that at least three different genes are present in the A. victoria population.
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Wavelength mutations and posttranslational autoxidation of green fluorescent protein
TL;DR: The availability of two visibly distinct colors should significantly extend the usefulness of GFP in molecular and cell biology by enabling in vivo visualization of differential gene expression and protein localization and measurement of protein association by fluorescence resonance energy transfer.
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Understanding, improving and using green fluorescent proteins.
Andrew B. Cubitt,Roger Heim,Stephen Adams,Aileen E. Boyd,Larry A. Gross,Roger Y. Tsien,Roger Y. Tsien +6 more
TL;DR: The GFP originally cloned from the jellyfish Aequorea victoria has several nonoptimal properties including low brightness, a significant delay between protein synthesis and fluorescence development, and complex photoisomerization, but can be re-engineered by mutagenesis to ameliorate these deficiencies and shift the excitation and emission wavelengths, creating different colors and new applications.