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Engineering 'Golden' fluorescentie door selectieve druk opneming van niet-canonieke aminozuren en eiwit analyse door massaspectrometrie en fluorescentie
JoVE Journal
Bioengineering
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JoVE Journal Bioengineering
Engineering ‘Golden’ Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
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Engineering 'Golden' fluorescentie door selectieve druk opneming van niet-canonieke aminozuren en eiwit analyse door massaspectrometrie en fluorescentie

Nederlands

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11,639 Views

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11:51 min

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April 27, 2018

DOI:

10.3791/57017-v

DOI:
10.3791/57017-v

•

11:51 min
April 27, 2018

•

1 Views
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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

Summary

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Synthetische biologie waardoor de engineering van eiwitten met ongekende eigenschappen met behulp van het co-translational inbrengen van niet-canonieke aminozuren. Hier, presenteerden we hoe een spectraal rood-verschoven variant van een GFP-type fluorophore met roman fluorescentie spectroscopische eigenschappen, genoemd "goud" fluorescent proteïne (GdFP), wordt geproduceerd in E. coli via selectieve druk opneming (SPI).

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Baumann, T., Schmitt, F., Pelzer, A., Spiering, V. J., Freiherr von Sass, G. J., Friedrich, T., Budisa, N. Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence. J. Vis. Exp. (134), e57017, doi:10.3791/57017 (2018).

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