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Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
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Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

DOI:
10.3791/56302-v

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11:25 min

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October 04, 2017

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, , , , ,
1Department of Physiology and Pharmacology, Schulich School of Medicine and Dentistry,University of Western Ontario

Capítulos

  • 00:05Título
  • 01:09Polymerase Chain Reaction (PCR)-mediated Site-directed Mutagenesis
  • 03:44Uniform 15N-labeled Protein Expression in BL21 ΔE3 E. coli
  • 05:24Purification of Recombinant Protein from E. coli
  • 07:50Chemical Attachment of Glucose-5-MTS to Protein by Dialysis
  • 08:48Results: Site-selective Cys Mutation Followed by In Vitro Glycosylation in STIM1 EF-SAM Protein
  • 10:11Conclusion

Summary

Tadução automática

Tadução automática

Biochemical and structural analyses of glycosylated proteins require relatively large amounts of homogeneous samples. Here, we present an efficient chemical method for site-specific glycosylation of recombinant proteins purified from bacteria by targeting reactive Cys thiols.

Tags

Keywords: Cysteine ThiolsSite-specific GlycosylationRecombinant ProteinsStructural BiologyGlycosylation PatternsDisease TherapyPCR MutagenesisDPN1 DigestionProtein Expression

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