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Inriktning cystein tioler för in Vitro platsspecifika Glycosylation av rekombinanta proteiner
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Targeting Cysteine Thiols for in Vitro Site-specific Glycosylation of Recombinant Proteins
DOI:

11:25 min

October 04, 2017

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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

Biokemiska och strukturella analyser av glykosylerat proteiner kräver relativt stora mängder homogena prover. Här presenterar vi en effektiv kemisk metod för platsspecifika glycosylation av rekombinanta proteiner renas från bakterier genom att rikta reaktiva Cys tioler.

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