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Målretning cystein dithioler for in Vitro lokationsspecifikke glykosylering af rekombinante proteiner
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Immunology and Infection
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JoVE Journal Immunology and Infection
Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins

Målretning cystein dithioler for in Vitro lokationsspecifikke glykosylering af rekombinante proteiner

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

Chapters

  • 00:05Title
  • 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

Automatic Translation

Automatic Translation

Biokemiske og strukturelle analyser af glykosyleret proteiner kræver relativt store mængder af homogene prøver. Her præsenterer vi en effektiv kemisk metode for lokationsspecifikke glykosylering af rekombinante proteiner renset fra bakterier ved at målrette reaktive Cys dithioler.

Tags

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

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