Red fluorescent protein

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Red fluorescent protein (RFP) is a fluorophore that fluoresces red-orange when excited. Several variants have been developed using directed mutagenesis.[1] The original was isolated from Discosoma, and named DsRed. Others are now available that fluoresce orange, red, and far-red.[2]

RFP is approximately 25.9 kDa. The excitation maximum is 558 nm, and the emission maximum is 583 nm.[3]

The first fluorescent protein to be discovered, green fluorescent protein (GFP), has been adapted to identify and develop fluorescent markers in other colors. Variants such as yellow fluorescent protein (YFP) and cyan fluorescent protein (CFP) were discovered in Anthozoa.[4]

Issues with fluorescent proteins include the length of time between protein synthesis and expression of fluorescence. DsRed has an maturation time of around 24 hours,[1] which can make it unusable for many experiments that take place in a shorter time frame. Additionally, DsRed exists in a tetrameric form, which can affect the function of proteins to which it is attached. Genetic engineering has improved the utility of RFP by increasing the speed of fluorescent development and creating monomeric variants.[3][4] Improved variants of RFP include mFruits (mCherry, mOrange, mRaspberry), mKO, TagRFP, mKate, mRuby, FusionRed, mScarlet and DsRed-Express.[4][5]

References[edit]

  1. ^ a b Bevis, Brooke J.; Glick, Benjamin S. (2002). "Rapidly maturing variants of the Discosoma red fluorescent protein (DsRed)". Nature Biotechnology. 20 (1): 83–87. doi:10.1038/nbt0102-83. ISSN 1546-1696. PMID 11753367.
  2. ^ Miyawaki, Atsushi; Shcherbakova, Daria M; Verkhusha, Vladislav V (October 2012). "Red fluorescent proteins: chromophore formation and cellular applications". Current Opinion in Structural Biology. 22 (5): 679–688. doi:10.1016/j.sbi.2012.09.002. ISSN 0959-440X. PMC 3737244. PMID 23000031.
  3. ^ a b Remington, S. James (1 January 2002). "Negotiating the speed bumps to fluorescence". Nature Biotechnology. 20 (1): 28–29. doi:10.1038/nbt0102-28.
  4. ^ a b c Piatkevich, Kiryl D.; Verkhusha, Vladislav V. (2011). "Guide to Red Fluorescent Proteins and Biosensors for Flow Cytometry". Methods in Cell Biology. 102: 431–461. doi:10.1016/B978-0-12-374912-3.00017-1. ISBN 9780123749123. ISSN 0091-679X. PMC 3987785. PMID 21704849.
  5. ^ Bindels, Daphne S; Haarbosch, Lindsay (2017). "mScarlet: a bright monomeric red fluorescent protein for cellular imaging". Nature Methods. 14 (1): 53–56. doi:10.1038/nmeth.4074. ISSN 1548-7105. PMID 27869816.