Expression of feline recombinant interferon-gamma in baculovirus and demonstration of biological activity

D J Argyle, M Harris, C Lawrence, K McBride, R Barron, C McGillivray, D E Onions

Research output: Contribution to journalArticlepeer-review

Abstract

We have previously reported the cloning of the coding sequence for feline-specific interferon-gamma. Here, we describe the expression of this sequence in a baculovirus system and demonstrate the biological activity of the recombinant protein. The coding sequence for feline interferon was directionally cloned into the baculovirus transfer vector pAcCL29-1. Transfer vector and linearized wild-type AcMNPV (BacPAK6) were used to co-transfect Sf9 cells by calcium phosphate coprecipitation. Subsequently, wild-type and recombinant viruses were separated by plaque assay. Recombinant plaques were expanded and a master stock of virus is produced. Production of biologically active interferon-gamma from infected Sf9 cells was demonstrated using a standard cytopathic effect reduction assay, utilising vesicular stomatitis virus (VSV), and an MHC class II induction assay.
Original languageEnglish
Pages (from-to)97-105
Number of pages9
JournalVeterinary Immunology and Immunopathology
Volume64
Issue number2
Publication statusPublished - 8 Jul 1998

Keywords

  • Animals
  • Cats
  • Cell Line
  • Cloning, Molecular
  • Cytopathogenic Effect, Viral
  • Gene Expression
  • Genetic Vectors
  • Histocompatibility Antigens Class II
  • Interferon-gamma
  • Nucleopolyhedrovirus
  • Recombinant Proteins
  • Spodoptera
  • Vesicular stomatitis Indiana virus

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