How is gfp used in research

Quantitative measurement of green fluorescent protein expression. Google Scholar. Allers, T. Improved strains and plasmid vectors for conditional overexpression of His-tagged proteins in Haloferax volcanii. Development of additional selectable markers for the halophilic archaeon Haloferax volcanii based on the leuB and trpA genes. Arpino, J. Crystal structure of enhanced green fluorescent protein to 1. PLoS One 7:e Baffour-Awuah, N. Bitan-Banin, G. Development of a gene knockout system for the halophilic archaeon Haloferax volcanii by use of the pyrE gene.

Brenneis, M. Experimental characterization of Cis-acting elements important for translation and transcription in halophilic archaea. PLoS Genet. Cho, S. Choi, J. The mechanism of folding robustness revealed by the crystal structure of extra-superfolder GFP.

FEBS Lett. Cline, S. Transformation of the archaebacterium Halobacterium volcanii with genomic DNA. Cormack, B. Gene , 33— Cubitt, A.

Understanding structure-function relationships in the Aequorea victoria green fluorescent protein. Methods Cell Biol. Danner, S. Characterization of the distal promoter element of halobacteria in vivo using saturation mutagenesis and selection.

Deschamps, J. Rapid purification of recombinant green fluorescent protein using the hydrophobic properties of an HPLC size-exclusion column. Protein Expr. Englert, C. Three different but related gene clusters encoding gas vesicles in halophilic archaea. Functional analysis of the gas vesicle gene cluster of the halophilic archaeon Haloferax mediterranei defines the vac-region boundary and suggests a regulatory role for the gvpD gene or its product.

Enoki, S. Acid denaturation and refolding of green fluorescent protein. Biochemistry 43, — Falb, M. Metabolism of halophilic archaea. Extremophiles 12, — Gregor, D. Use of a halobacterial bgaH reporter gene to analyse the regulation of gene expression in halophilic archaea. Microbiology Pt 7 , — Griesbeck, O. Reducing the environmental sensitivity of yellow fluorescent protein. Mechanism and applications. Hofacker, A. GvpE- and GvpD-mediated transcription regulation of the p- gvp genes encoding gas vesicles in Halobacterium salinarum.

Microbiology , — Holmes, M. Improved shuttle vectors for Haloferax volcanii including a dual-resistance plasmid. Gene , — Horne, M. A DNA region of 9 kbp contains all genes necessary for gas vesicle synthesis in halophilic archaebacteria.

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Jackson, S. Understanding the folding of GFP using biophysical techniques. Proteomics 3, — Kerscher, L. Kimata, Y. A novel mutation which enhances the fluorescence of green fluorescent protein at high temperatures. Kremers, G. Fluorescent proteins at a glance. Cell Sci. Lam, W. Shuttle vectors for the archaebacterium Halobacterium volcanii.

Lissemore, J. Green fluorescent protein as a quantitative reporter of relative promoter activity in E. Biotechniques , 88— Marschaus, L. A dual promoter region with overlapping activator sequences drives the expression of gas vesicle protein genes in haloarchaea. Nomura, S. GFP does not interfere with any biological processes. Transgenic mice can be labeled with GFP, which is then easily observed in their offspring just by exposing them to blue or UV light, as seen in Fig.

GFP can be fused to other proteins, effectively making those proteins fluorescent. This allows any protein to be localized and tracked using standard fluorescent microscopy, by shining a blue light on the cells, the protein of interest will fluoresce back with a green light. GFP in live-cell experiments : the classic green fluorescent molecule is fluorescein isothiocyanate FITC , but this is toxic to cells and cannot be used directly without first fixing the cells or causing unavoidable damage.

GFP is far less harmful as it is a naturally-occurring protein and can be used in experiments on live cells while causing virtually no damage, especially if it is passed on to offspring. GFP in advanced microscopy applications. GFP is modifiable , as the genetic and amino acid code for GFP is well understood it has been subject to several modifications.

Firstly GFP was modified to produce enhanced GFP eGFP , which has increased fluorescence intensity, greater photostability, more convenient excitation peaks and higher efficiency at room temperature.

Due to Tsien's and other bioengineers' efforts, GFP could not only exhibit brighter fluorescence, but also respond to a wider range of wavelengths, as well as emit almost all colors, except for red. Tsien's findings enabled scientists to tag multiple colored GFPs to different proteins, cells, or organelles of interest, and scientists could study the interaction of those particles. Other laboratories developed fluorescent sensors for calcium, protease and other biological molecules.

Since then, scientists have reported more than distinct GFP-like proteins in many species. As GFP does not interfere with biological processes when used in vivo , biologists use it to study how organisms develop.

For example, after , Chalfie and his colleagues applied GFP in the study of the neuron development of C. In a paper, Chalfie and his colleagues describe how they first labeled a specific gene involved in tactile perception in neuron cells with GFP, and then observed the amount of fluorescence emitted by those cells.

Because mutant cells produced less or more GFP than normal cells, the abnormal amount of fluorescence production indicated the abnormal development of mutants. Since then, this field of research expanded to many other organisms, including fruitflies, mice, and zebra fish.

Green Fluorescent Protein Green fluorescent protein GFP is a protein in the jellyfish Aequorea Victoria that exhibits green fluorescence when exposed to light. Ward, and Douglas C. Chalfie, Martin. Davenport, Demorest and Joseph Nicol. Harvey, Edmund. Luminescence in the coelenterates. Hastings, John, and James Morin. Matz, Mikhail V.

Fradkov, Yulii A. Labas, Aleksandr P. Savitsky, Andrey G. Plasmids of this type may have the GFP under the control of an additional promoter from that of the gene of interest, or expressed from the same transcript as the gene of interest, but after an internal ribosome entry site IRES. This is oftentimes used in conjuction with FACS see below. Fluorescence-activated cell sorting FACS : This is a type of flow cytometry that separates mixtures of cells into distinct populations based on fluorescent signal.

It can also be used, when put under control of promoters of interest, to visualize the developmental stage at which these promoters are active. Further, GFP can label transgenically modified ES cells, which can then be used for implantation and generation of transgenic mice. Purification: GFP can be used as a general epitope tag for protein purification and a number of commercial antibodies to GFP are available. Do you have a favorite way to use GFP? Comment and tell us how!

Sharing science just got easier Subscribe to our blog. Follow Addgene on Social. Increased fluorescence, photostability, and a shift of the major excitation peak to nm.

Causes the chromophore to form with an indole rather than phenol component cyan derivatives.