Extraction, Purification and Properties of Aequorin, a Bioluminescent Protein from the Luminous Hydromedusan, <i>Aequorea</i>

Shimomura, Osamu; Johnson, Frank H.; Saiga, Yo

doi:10.1002/jcp.1030590302

Cited by 2,050 publications

(1,049 citation statements)

References 23 publications

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“…Green fluorescent protein (GFP) was originally isolated from the jellyfish Aequorea victoria [5]. It consists of 238 amino acids with a molecular weight of 26.9 kDa.…”

Section: Biological and Genetic Backgroundmentioning

confidence: 99%

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Teaching bioprocess engineering to undergraduates: Multidisciplinary hands‐on training in a one‐week practical course

Henkel

Zwick

Beuker

et al. 2015

Biochem Molecular Bio Educ

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Bioprocess engineering is a highly interdisciplinary field of study which is strongly benefited by practical courses where students can actively experience the interconnection between biology, engineering, and physical sciences. This work describes a lab course developed for 2nd year undergraduate students of bioprocess engineering and related disciplines, where students are challenged with a real-life bioprocess-engineering application, the production of recombinant protein in a fed-batch process. The lab course was designed to introduce students to the subject of operating and supervising an experiment in a bioreactor, along with the analysis of collected data and a final critical evaluation of the experiment. To provide visual feedback of the experimental outcome, the organism used during class was Escherichia coli which carried a plasmid to recombinantly produce enhanced green fluorescent protein (eGFP) upon induction. This can easily be visualized in both the bioreactor and samples by using ultraviolet light. The lab course is performed with bioreactors of the simplest design, and is therefore highly flexible, robust and easy to reproduce. As part of this work the implementation and framework, the results, the evaluation and assessment of student learning combined with opinion surveys are presented, which provides a basis for instructors intending to implement a similar lab course at their respective institution.

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“…Green fluorescent protein (GFP) was originally isolated from the jellyfish Aequorea victoria [5]. It consists of 238 amino acids with a molecular weight of 26.9 kDa.…”

Section: Biological and Genetic Backgroundmentioning

confidence: 99%

“…(5)]. The saturation concentration was provided by the instructor (7 mg L 21 ), which can be approximated by referring to saturation concentrations of oxygen in a watersaline system at 37 C or measured by O 2 -titration.…”

Section: Calculationsmentioning

confidence: 99%

Teaching bioprocess engineering to undergraduates: Multidisciplinary hands‐on training in a one‐week practical course

Henkel

Zwick

Beuker

et al. 2015

Biochem Molecular Bio Educ

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“…Fluorescent proteins such as green fluorescent protein (GFP), whose discovery and development led to the award of the Nobel Prize in Chemistry in 2008, are the most useful visual markers (Shimomura et al 1962). Fluorescent proteins enable direct observation and an easier and more precise evaluation of transformation events in real time under living conditions, without treatment with any exogenous substrate.…”

Section: Introductionmentioning

confidence: 99%

Visual selection allows immediate identification of transgenic rice calli efficiently accumulating transgene products

Saika

Toki

2009

Plant Cell Rep

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In genetic transformation systems, antibiotic resistance genes are routinely used as powerful markers for selecting transformed cells from surrounding non-transformed cells. However, simultaneous use of the gene encoding green fluorescent protein (GFP) and an antibiotic resistance gene facilitates the selection process, since it allows visible selection of transformed cells. Here, we report the development of a visual selection system for transformed cells using a GFP marker without selection against antibiotics after Agrobacterium-mediated transformation in rice. Both GFP protein levels and GFP fluorescence in calli isolated by visual selection were higher than in calli selected on hygromycin (Hyg), suggesting that transgenic calli hyper-accumulating GFP were efficiently obtained by selection using GFP fluorescence itself rather than Hyg resistance. Furthermore, gfp transcripts in calli isolated by visual selection were more abundant than under Hyg selection; in contrast, transcript levels of hpt in calli selected visually were comparable to those obtained under Hyg selection. These results suggest that there was no correlation between hpt and gfp expression levels, despite the fact that they are aligned in tandem on an integrated locus after selection by either GFP fluorescence or Hyg resistance. This fact indicates that positional effects can influence the expression of each transgene differently, even when they are located in tandem at the same locus. In summary, based on our results, we discuss a model system for rice cell culture transformation for the production of recombinant proteins using visual selection.

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“…Although Shimomura initially isolated a blue luminescent protein (termed 'aequorin') [2], it later became clear that the green glow of the jellyfish is due to the presence of an inherently fluorescent protein, later termed as green fluorescent protein. Two major developments in the early 1990's dramatically changed the course of future GFP research and revolutionized modern cell biology: (i) Cloning of the gene that encodes GFP by Douglas Prasher, Franklyn Prendergast and co-workers [3]; and (ii) successful demonstration by Martin Chalfie, Douglas Prasher and co-workers that GFP could be expressed in heterologous systems such as E. coli and C. elegans [4].…”

mentioning

confidence: 99%

The Green Journey

Haldar

Chattopadhyay

2008

J Fluoresc

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Extraction, Purification and Properties of Aequorin, a Bioluminescent Protein from the Luminous Hydromedusan, Aequorea

Cited by 2,050 publications

References 23 publications

Teaching bioprocess engineering to undergraduates: Multidisciplinary hands‐on training in a one‐week practical course

Teaching bioprocess engineering to undergraduates: Multidisciplinary hands‐on training in a one‐week practical course

Visual selection allows immediate identification of transgenic rice calli efficiently accumulating transgene products

The Green Journey

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