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Genetic Incorporation of Biosynthesized L-dihydroxyphenylalanine (DOPA) and Its Application to Protein Conjugation
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Biochemistry
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JoVE Journal Biochemistry
Genetic Incorporation of Biosynthesized L-dihydroxyphenylalanine (DOPA) and Its Application to Protein Conjugation

Genetic Incorporation of Biosynthesized L-dihydroxyphenylalanine (DOPA) and Its Application to Protein Conjugation

DOI:
10.3791/58383-v

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10:24 min

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August 24, 2018

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,
1Department of Chemistry,Sogang University

Chapters

  • 00:04Title
  • 00:29Culture Preparation
  • 02:06Expression and Purification of GFP-E90DOPA by a Biosynthetic System
  • 04:25Oligomerization of Purified GFP-E90DOPA and SDS-PAGE Analysis
  • 05:47Conjugation of GFP-E90DOPA with an Alkyne Probe by SPOCQ, Purification of the Labeled GFP
  • 07:10SDS-PAGE Analysis and Fluorescence Gel Scanning of Labeled GFP
  • 08:21Results: Genetic Incorporation of Biosynthesized L-dihydroxyphenylalanine
  • 10:01Conclusion

Summary

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Automatic Translation

Here, we present a protocol for the genetic incorporation of L-dihydroxyphenylalanine biosynthesized from simple starting materials and its application to protein conjugation.

Tags

Keywords: Genetic IncorporationL-dihydroxyphenylalanine (DOPA)Protein ConjugationBiosynthetic SystemE. ColiExpression PlasmidElectroporationLB MediumPA-5052 MediumCell LysisProtein Purification

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