Reporter

Part:BBa_K3165019

Designed by: Mohit Das   Group: iGEM19_IISc-Bangalore   (2019-10-19)
Revision as of 11:09, 19 October 2019 by Mohitdas (Talk | contribs)


mOrange v 2.0

mOrange is an mRFP derived fluorescent protein used as a reporter in various biological studies. Due to its high quantum yield and sufficiently large extinction coefficient, it produces a bright mature protein (an important attribute of reporter proteins)


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Usage and Biology

Biology

mRFP is a monomeric fluorescent protein artificially derived from the naturally occurring DsRed protein synthesized by Discosoma sp. To overcome the toxicity of the tetrameric DsRed, a monomeric form, mRFP was developed which quickly became popular as a reliable fluorophore. Since the introduction of mRFP, an enormous range of protein has come into existence by specific mutations in the mRFP sequence. mFruits is a class of fluorescent proteins artificially derived from mRFP which host a range of fluorophores with varying excitation and emission wavelengths. mOrange due to its high quantum yield and respectably large excitation coefficient is one of the brightest true monomers among the mRFP derivatives. It was synthesised after six rounds of directed evolution from mRFP. Although the protein has a slightly longer maturation time than mCherry, it exhibits substantial acid sensitivity which makes it a favourable choice for various biological studies.

Usage

mOrange is used as a non-invasive reporter for gene expression. It can be placed under a gene regulatory system (like the optogenetic T7 RNA Polymerase system) to characterise the functioning of the system via fluorescence analysis.

References :

(1) Shaner, N. C., Campbell, R. E., Steinbach, P. A., Giepmans, B. N. G., Palmer, A. E., & Tsien, R. Y. (2004). Improved monomeric red, orange and yellow fluorescent proteins derived from Discosoma sp. red fluorescent protein. Nature Biotechnology, 22(12), 1567–1572. doi:10.1038/nbt1037 


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