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Rescue of Infectious Salmon Anemia Virus (ISAV) from Cloned cDNA

  • Daniela Toro-Ascuy
  • Marcelo Cortez-San MartínEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1602)

Abstract

The piscine Orthomyxovirus called Infectious Salmon Anemia Virus (ISAV) is one of the most important emerging pathogens affecting the salmon industry worldwide. The first reverse genetics system for ISAV, which allows the generation of recombinant ISA virus (rISAV), is an important tool for the characterization and study of this fish virus. The plasmid-based reverse genetics system for ISAV includes the use of a novel fish promoter, the Atlantic salmon internal transcribed spacer region 1 (ITS-1). The salmon, viral and mammalian genetic elements included in pSS-URG vectors allow the expression of the eight viral RNA segments. In addition to four cytomegalovirus (CMV)-based vectors that express the four proteins of the ISAV ribonucleoprotein complex, the eight pSS-URG vectors allowed the generation of infectious rISAV in salmon cells.

Key words

Infectious Salmon Anemia Virus ISAV Viral RNA Reverse genetics Salmon cells Transfection ITS-1 RNA polymerase I and RNA polymerase II 

Notes

Acknowledgements

We thank Prof. Daniel R. Perez for technical and scientific support during ISAV reverse genetics development. We are grateful to Dee Sagawe for proofreading the manuscript. We acknowledge financial support Fondecyt 11110212, Fondecyt 1161006, VIU 14E025. M.C.-S.M. thanks to DICYT-USACH and DGT-USACH. D.T.-A. thanks Conicyt-Chile for a PhD fellowship.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Daniela Toro-Ascuy
    • 1
  • Marcelo Cortez-San Martín
    • 1
    Email author
  1. 1.Laboratory of Molecular Virology, Faculty of Chemistry and BiologyUniversity of Santiago of ChileSantiagoChile

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