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Generation and Assembly of Virus-Specific Nucleocapsids of the Respiratory Syncytial Virus
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生物化学
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JoVE 杂志 生物化学
Generation and Assembly of Virus-Specific Nucleocapsids of the Respiratory Syncytial Virus

Generation and Assembly of Virus-Specific Nucleocapsids of the Respiratory Syncytial Virus

DOI:
10.3791/62010-v

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09:08 min

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July 27, 2021

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, , , ,
1Department of Biochemistry,Emory University School of Medicine

Chapters

  • 00:04Introduction
  • 01:10Protein Expression and Purification
  • 04:12In Vitro Virus-Specific Nucleocapsid (NC) Assembly
  • 05:19Negative Stain Grid Preparation
  • 07:17Results: Representative N1-391P1-126 Complex Copurification
  • 08:08Conclusion

Summary

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For in-depth mechanistic analysis of the respiratory syncytial virus (RSV) RNA synthesis, we report a protocol of utilizing the chaperone phosphoprotein (P) for coexpression of the RNA-free nucleoprotein (N0) for subsequent in vitro assembly of the virus-specific nucleocapsids (NCs).

Tags

Virus-specific NucleocapsidsRespiratory Syncytial VirusRecombinant Viral Protein ExpressionRNA-free N ProteinLigation Independent CloningElectron MicroscopyBi-cistronic ConstructionProtein ExpressionChromatographyCobalt Gravity ColumnQ ColumnHPLCUV Detection

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