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 language | English |
|---|---|
| Pages (from-to) | 97-105 |
| Number of pages | 9 |
| Journal | Veterinary Immunology and Immunopathology |
| Volume | 64 |
| Issue number | 2 |
| Publication status | Published - 8 Jul 1998 |
Keywords / Materials (for Non-textual outputs)
- 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