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Produktion af Menneskelige Norovirus fremspringende domæner i<em> E. coli</em> Til røntgenkrystallografi
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JoVE Journal Biologia
Production of Human Norovirus Protruding Domains in E. coli for X-ray Crystallography

Produktion af Menneskelige Norovirus fremspringende domæner i<em> E. coli</em> Til røntgenkrystallografi

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

Capítulos

  • 00:05Título
  • 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

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Tadução automática

Her beskriver vi en fremgangsmåde til at udtrykke og oprense kvalitet norovirus høj udragende (P) domæner i E. coli til anvendelse i røntgenkrystallografi undersøgelser. Denne metode kan anvendes på andre calicivirus P domæner, såvel som ikke-strukturelle proteiner, dvs.., Viralt protein genom-linked (VPG), protease, og RNA-afhængige 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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