Primary structure of the Aequorea victoria green-fluorescent protein

Prasher, Douglas C.; Eckenrode, Virginia K.; Ward, William W.; Prendergast, F.G.; Cormier, Milton J.

doi:10.1016/0378-1119(92)90691-h

Cited by 1,960 publications

(1,081 citation statements)

References 16 publications

Supporting

Mentioning

1,036

Contrasting

Unclassified

Order By: Relevance

“…E-mail: hhgerdes@sun0.urz.uni-heidelberg.de Abbreviations: GFP, green fluorescent protein; BFP, blue fluorescent protein; TGN, trans-Golgi network; ER, endoplasmic reticulum; PM, plasma membrane 30 kDa on SDS-PAGE [7]. Its chromophore is formed by cyclisation and oxidation of the three amino acids Ser65, Tyr66 and Gly67 [3,8]. This posttranslational modification (maturation) occurs within 2 to 4 h after synthesis and is probably autocatalytic [8].…”

Section: Properties Of Wildtype and Mutant Gfpsmentioning

confidence: 99%

“…This GFP has a unique property in that it forms a chromophore of three amino acids within its primary structure and, in contrast to other bioluminescent molecules, operates independently of cofactors [1]. Although the properties of the protein have been known for many years [2], it was not until the cloning of the cDNA of GFP in 1992 [3] and its subsequent heterologous expression in E. coli and C. elegans [4] that researchers from many fields became aware of the potential of this molecule. Up to this point the work of many laboratories had shown that GFP can produce green fluorescence in a variety of diverse organisms such as bacteria, slime molds, plants and mammals [2].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Green fluorescent protein: applications in cell biology

1996

View full text Add to dashboard Cite

The green fluorescent protein (GFP) of Aequorea victoria is a unique in vivo reporter for monitoring dynamic processes in cells or organisms. As a fusion tag GFP can be used to localize proteins, to follow their movement or to study the dynamics of the subcellular compartments to which these proteins are targeted. Recent studies where GFP technology has revealed new insights regarding physiological activities of living cells are discussed.

show abstract

Section: Properties Of Wildtype and Mutant Gfpsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Green fluorescent protein: applications in cell biology

1996

View full text Add to dashboard Cite

show abstract

“…Its X-ray barrel-type structure was resolved at 1.85Å (PDB entry: 1W7S; [6]), and is formed by eleven β-sheets and one α-helix, to which the GFP chromophore is connected. The latter is obtained by a posttranslational cyclization reaction of the polypeptide skeleton Ser65, Tyr66 and Gly67 residues, followed by oxidation of the Tyr66 residue lateral side-chain [7][8][9]. Through a hydrogen bonds network involving particular polar residues and H 2 O molecules, the cromophore is able to establish noncovalent interactions with the protein [10,11].…”

Section: Introductionmentioning

confidence: 99%

Thermal Effect on Aequorea Green Fluorescent Protein Anionic and Neutral Chromophore Forms Fluorescence

Santos

2011

J Fluoresc

View full text Add to dashboard Cite

The emission behaviour of Aequorea green fluorescent protein (A-GFP) chromophore, in both neutral (N) and anionic (A) form, was studied in the temperature range from 20°C to 75°C and at pH=7. Excitation wavelengths of 399 nm and 476 nm were applied to probe the N and A forms environment, respectively. Both forms exhibit distinct fluorescence patterns at high temperature values. The emission quenching rate, following a temperature increase, is higher for the chromophore N form as a result of the hydrogen bond network weakening. The chromophore anionic form emission maximum is red shifted, upon temperature increase, due to a charge transfer process occurring after A form excitation.

show abstract

“…[1][2][3] procedures, a feature fulfilled by green fluorescent protein. Advantages include the ability to produce high-titer stocks, the high efficiency of gene transfer into a variety Originally isolated from the jellyfish Aquorea victoria as a 238 amino acid protein, 15 wild-type GFP emits a bright of cell types, and the ability to transduce cells that have a low mitotic index, eg the endothelium, kidney and cells green fluorescence upon excitation at both 395 and 475 nm. Crystallization has revealed a cylinder-like structure of the central nervous system.…”

mentioning

confidence: 99%

Adenovirus-mediated expression of green fluorescent protein

et al. 1997

View full text Add to dashboard Cite

A recombinant replication-deficient adenovirus has been blotting. As demonstrated by FACS analysis, up to 98% of generated that expresses a mutant of the Aquorea victoria HUVEC and approximately 70% of human smooth muscle green fluorescent protein (GFP) under the control of the cells could be transduced to express GFP. Since GFP can strong CMV promoter by insertion into the E1 region (AdVbe detected in cells without the need for prior fixing and GFP). High expression of GFP was found in different cell staining, the virus should be useful for optimizing in living types after infection with the recombinant virus that could cells the transduction efficiency of different cell types, of be easily detected by fluorescence microscopy. In human cells from different experimental animals, as well as studyumbilical vein endothelial cells (HUVEC), expression levels ing the kinetics and persistence of adenovirus-mediated had already reached a maximum after 2 days and were gene transfer in diverse experimental settings. stable for at least 7 days, as determined by Western Keywords: green fluorescent protein; recombinant adenovirus; marker gene; endothelium Gene transfer techniques using replication-deficient of them can be directly visualized in living cells without the need for prior incubation with substrates or staining recombinant adenoviral vectors (AdV) have considerable potential both at experimental and therapeutic levels. [1][2][3] procedures, a feature fulfilled by green fluorescent protein. Advantages include the ability to produce high-titer stocks, the high efficiency of gene transfer into a variety Originally isolated from the jellyfish Aquorea victoria as a 238 amino acid protein, 15 wild-type GFP emits a bright of cell types, and the ability to transduce cells that have a low mitotic index, eg the endothelium, kidney and cells green fluorescence upon excitation at both 395 and 475 nm. Crystallization has revealed a cylinder-like structure of the central nervous system. 4,5 Limitations of AdV include the relatively short time of expression of the of ␤-sheets wrapped around a single central helix. 16 The fluorophore results from the autocatalytic cyclization (therapeutic) gene in vivo (a few weeks to several months, depending on the organ) due to the episomal maintebetween amino acids of the polypeptide backbone and oxidation of the ␣-␤ bond of the central tyrosine. Due to nance of the adenoviral DNA in the cell, and the elicitation of an immune response by the host organism. [6][7][8] this rigid encapsulation, the fluorophore is remarkably insensitive to changes in environmental conditions, eg These features favor, in the first instance, applications where only a transient expression of the ectopic gene is pH, O 2 quenching of the excited state, and to proteolysis. The generation of mutants that show improved levels of desirable, eg inflammatory reactions.Despite its broad cell-type specificity, considerable diffluorescence as well as different excitation and emission maxima has further triggered the widesprea...

show abstract

Primary structure of the Aequorea victoria green-fluorescent protein

Cited by 1,960 publications

References 16 publications

Green fluorescent protein: applications in cell biology

Green fluorescent protein: applications in cell biology

Thermal Effect on Aequorea Green Fluorescent Protein Anionic and Neutral Chromophore Forms Fluorescence

Adenovirus-mediated expression of green fluorescent protein

Contact Info

Product

Resources

About