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Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography
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Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography

Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography

DOI:
10.3791/53845-v

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10:46 min

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April 19, 2016

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, , ,
1Schaller Research Group,University of Heidelberg and the German Cancer Research Center, 2Department of Infectious Diseases, Virology,University of Heidelberg

Chapters

  • 00:05Title
  • 00:47P Domain Expression
  • 02:251st Purification Step and Protease Cleavage
  • 05:142nd Purification Step
  • 06:283rd Purification Step
  • 07:28Crystallization of the P Domain
  • 09:01Results: Evaluation of Protein Elution
  • 09:46Conclusion

Summary

Automatic Translation

Automatic Translation

Here, we describe a method to express and purify high quality norovirus protruding (P) domains in E. coli for use in X-ray crystallography studies. This method can be applied to other calicivirus P domains, as well as non-structural proteins, i.e., viral protein genome-linked (VPg), protease, and RNA dependent RNA polymerase (RdRp).

Tags

Keywords: NorovirusProtruding DomainE. ColiX-ray CrystallographyProtein ExpressionProtein PurificationProtein CrystallizationMBP-His-P Domain Fusion ProteinTransformationIPTG InductionCell LysisNickel-NTA Agarose Purification

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