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Engineering 'Golden' fluorescens vid selektionstryck inkorporering av icke-kanoniska aminosyror och proteinanalys av masspektrometri och fluorescens
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1Institute of Chemistry L 1, Department of Biocatalysis,Technical University of Berlin, 2Institute of Chemistry PC 14, Department of Bioenergetics,Technical University of Berlin, 3Institute of Chemistry TC 7, Department of Physical Chemistry/Molecular Material Sciences,Technical University of Berlin
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
Syntetisk biologi möjliggör konstruktion av proteiner med oöverträffade egenskaper med hjälp av co-translational införande av icke-kanoniska aminosyror. Vi presenterade här, hur en spektralt röd-skiftat variant av en GFP-typ fluorophore med romanen fluorescens spektroskopiska egenskaper, kallas ”guld” fluorescerande protein (GdFP), produceras i E. coli via selektionstryck inkorporering (SPI).
