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Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins
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Imunologia e Infecção
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JoVE Journal Imunologia e Infecção
Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins

Simplified Reverse Genetics Method to Recover Recombinant Rotaviruses Expressing Reporter Proteins

DOI:
10.3791/61039-v

•

11:40 min

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April 17, 2020

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, , ,
1Department of Biology,Indiana University

Capítulos

  • 00:04Introduction
  • 00:32Generation of Recombinant Virus
  • 04:00Plaque Isolation of Recombinant Viruses
  • 07:02Gel Electrophoresis of Viral dsRNA
  • 09:11Results: Characteristics of the Recombinant Strains rSA11/wt and rSA11/NSP3-2A-3xFL-UnaG
  • 10:58Conclusion

Summary

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Generation of recombinant rotaviruses from plasmid DNA provides an essential tool for the study of rotavirus replication and pathogenesis, and the development of rotavirus expression vectors and vaccines. Herein, we describe a simplified reverse genetics approach for generating recombinant rotaviruses, including strains expressing fluorescent reporter proteins.

Tags

Keywords: Reverse GeneticsRecombinant RotavirusReporter ProteinsBHKT7 CellsMA104 CellsTransfectionTrypsinPlasmid MixtureFluorescent Marker Proteins

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