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Målretting cystein Thiols for in Vitro områdespesifikke glykosylering rekombinante 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

Biokjemiske og strukturelle analyser av glycosylated proteiner krever relativt store mengder homogen prøver. Her presenterer vi en effektiv kjemisk metode for områdespesifikke glykosylering rekombinante proteiner renset fra bakterier av målretting reaktive Cys thiols.

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