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Utilising a Defective IBV RNA for Heterologous Gene Expression with Potential Prophylactic Application

  • S. A. Evans
  • K. Stirrups
  • K. Dalton
  • K. Shaw
  • D. Cavanagh
  • P. Britton
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  • 31 Downloads
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 440)

Abstract

Based on the natural ability of coronaviruses to undergo homologous RNA recombination, we are working to produce infectious bronchitis virus (IBV) recombinants using RNA generated from recombinant fowlpox viruses (FPV). The aim is to replace the spike (S) gene of an existing IBV vaccine strain with the S gene of a heterologous strain. CD-61 is an IBV defective RNA (D-RNA) derived from a naturally occurring IBV D-RNA(CD-91). CD-61 D-RNA is being investigated as an RNA vector for the expression of heterologous genes. T7-derived RNA transcripts of CD-61 can be replicated and passaged in the presence of helper virus, following electroporation into IBV-infected cells. CD-61 cDNA was modified by the addition of the hepatitis delta virus ribozyme plus T7 terminator downstream of the 3’ UTR. This allowed the synthesis of discreet RNA transcripts. The complete cassette was cloned into an FPV transfer vector (pEFL10) for generating recombinant fowlpox viruses. FPV/CD-61 recombinants will be assessed for D-RNA production in IBV-infected cells. The luciferase reporter gene sequence has been inserted into the modified CD-61, under the control of the IBV transcription associated sequence (TAS) from gene 5. Luciferase has been successfully expressed from CD-61 in helper virus-infected cells.

Keywords

Infectious Bronchitis Virus Heterologous Gene Expression Infectious Bronchitis Virus Strain Chicken Embryo Fibroblast Cell Fowlpox Virus 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

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Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • S. A. Evans
    • 1
  • K. Stirrups
    • 1
  • K. Dalton
    • 1
  • K. Shaw
    • 1
  • D. Cavanagh
    • 1
  • P. Britton
    • 1
  1. 1.Division of Molecular Biology, Institute for Animal HealthCompton LaboratoryComptonUK

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