Optical properties of the monomeric red fluorescent protein mRFP1

Vrzheshch, E. P.; Dmitrienko, D. V.; Rudanov, G. S.; Zagidullin, V. E.; Paschenko, Vladimir Z.; Razzhivin, A. P.; Saletsky, A. M.; Vrzheshch, P. V.

doi:10.3103/s0096392508030036

Cited by 4 publications

(2 citation statements)

References 18 publications

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“…Eluted protein was dialyzed against 20 mM Tris-HCl, 150 mM NaCl, 5 mM 2-mercaptoethanol, pH 8. Protein concentrations were determined from the extinction coefficients of GFPmut3b (ε 501 nm = 21,890 M −1 cm −1 ), 35 mCerulean (ε 433 nm = 43 000 M −1 cm −1 ), 35 and mRFP (ε 584 nm = 44 000 M −1 cm −1 ) 36 with an Agilent 8453 UV−vis. Spinach mRNA was purified by using acidified phenol extraction followed by an ethanol precipitation.…”

Section: Spinach and Fluorescent Proteins Allow For The Simultaneous ...mentioning

confidence: 99%

Gene Position More Strongly Influences Cell-Free Protein Expression from Operons than T7 Transcriptional Promoter Strength

et al. 2013

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The cell-free transcription−translation of multiple proteins typically exploits genes placed behind strong transcriptional promoters that reside on separate pieces of DNA so that protein levels can be easily controlled by changing DNA template concentration. However, such systems are not amenable to the construction of artificial cells with a synthetic genome. Herein, we evaluated the activity of a series of T7 transcriptional promoters by monitoring the fluorescence arising from a genetically encoded Spinach aptamer. Subsequently the influences of transcriptional promoter strength on fluorescent protein synthesis from one, two, and three gene operons were assessed. It was found that transcriptional promoter strength was more effective at controlling RNA synthesis than protein synthesis in vitro with the PURE system. Conversely, the gene position within the operon strongly influenced protein synthesis but not RNA synthesis.

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Section: Spinach and Fluorescent Proteins Allow For The Simultaneous ...mentioning

confidence: 99%

Gene Position More Strongly Influences Cell-Free Protein Expression from Operons than T7 Transcriptional Promoter Strength

et al. 2013

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“…Advances in fluorescent protein technology and FP-based biosensors enabled the real-time monitoring of cellular and molecular events in live cells with high spatiotemporal resolutions [17][18][19]. Different sensing strategies of FP-based biosensors have been developed [20], in particular biosensors to monitor the progression of autophagy are based on different FP properties such as pH-sensitivity, stability, brightness, spectral profile, and fluorescence resonance energy transfer (FRET) [12,14,[21][22][23][24][25][26][27][28][29][30][31][32][33][34].…”

Section: Introductionmentioning

confidence: 99%

Fluorescent Protein-Based Autophagy Biosensors

Kim

2021

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Autophagy is an essential cellular process of self-degradation for dysfunctional or unnecessary cytosolic constituents and organelles. Dysregulation of autophagy is thus involved in various diseases such as neurodegenerative diseases. To investigate the complex process of autophagy, various biochemical, chemical assays, and imaging methods have been developed. Here we introduce various methods to study autophagy, in particular focusing on the review of designs, principles, and limitations of the fluorescent protein (FP)-based autophagy biosensors. Different physicochemical properties of FPs, such as pH-sensitivity, stability, brightness, spectral profile, and fluorescence resonance energy transfer (FRET), are considered to design autophagy biosensors. These FP-based biosensors allow for sensitive detection and real-time monitoring of autophagy progression in live cells with high spatiotemporal resolution. We also discuss future directions utilizing an optobiochemical strategy to investigate the in-depth mechanisms of autophagy. These cutting-edge technologies will further help us to develop the treatment strategies of autophagy-related diseases.

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Macro To Nano Spectroscopy

Uddin¹

2012

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Optical properties of the monomeric red fluorescent protein mRFP1

Cited by 4 publications

References 18 publications

Gene Position More Strongly Influences Cell-Free Protein Expression from Operons than T7 Transcriptional Promoter Strength

Gene Position More Strongly Influences Cell-Free Protein Expression from Operons than T7 Transcriptional Promoter Strength

Fluorescent Protein-Based Autophagy Biosensors

Macro To Nano Spectroscopy

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