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用质谱和荧光法研究非规范氨基酸和蛋白质分析的选择性压力下工程 "金" 荧光
JoVE Journal
Bioengineering
This content is Free Access.
JoVE Journal Bioengineering
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

Automatic Translation

合成生物学使蛋白质的工程具有空前的特性, 使用非规范氨基酸的共平移插入。在这里, 我们介绍了如何一个光谱红色转移变种的 GFP 型荧光与新的荧光光谱性质, 称为 "金" 荧光蛋白 (GdFP), 是产生于大肠杆菌通过选择性压力成立 (SPI)。

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