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Engineering ‘Golden’ Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
JoVE 신문
생체공학
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JoVE 신문 생체공학
Engineering ‘Golden’ Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
DOI:

11:51 min

April 27, 2018

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Chapters

  • 00:05Title
  • 01:46Recombinant Protein Expression
  • 04:05Target Protein Purification via Immobilized Metal Ion Affinity Chromatography
  • 05:20Intact Protein Mass Analysis by High-performance Liquid Chromatography Coupled to Electrospray Ionization Time-of-flight Mass Spectrometry
  • 06:38Fluorescence Lifetime Measurements and Decay-associated Spectra of GdFP
  • 07:59Results: Gold Fluorescent Protein Analysis by Mass Spectrometry and Fluorescence
  • 10:13Conclusion

Summary

자동 번역

Synthetic biology enables the engineering of proteins with unprecedented properties using the co-translational insertion of non-canonical amino acids. Here, we presented how a spectrally red-shifted variant of a GFP-type fluorophore with novel fluorescence spectroscopic properties, termed "gold" fluorescent protein (GdFP), is produced in E. coli via selective pressure incorporation (SPI).

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