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Expression of a bioactive bovine interleukin-12 using baculovirus
K Takehara1, T Nagata, R Kikuma
1Laboratory of Poultry Diseases, School of Veterinary Medicine and Animal Sciences, Kitasato University, Towada, 034-8628, Aomori, Japan. takehara@vmas.kitasato-u.ac.jp
Veterinary Immunology and Immunopathology
|November 9, 2000
Summary
Researchers developed a baculovirus system to produce bioactive bovine interleukin-12 (bovine IL-12). This method enables large-scale production of functional bovine IL-12 for potential therapeutic applications.
Area of Science:
- * Molecular Biology
- * Immunology
- * Virology
Background:
- * Bovine interleukin-12 (bovine IL-12) is a critical cytokine for immune responses.
- * Efficient and scalable production of bioactive bovine IL-12 is essential for research and potential therapeutic use.
Purpose of the Study:
- * To construct recombinant baculoviruses for expressing bovine IL-12 subunits (p35 and p40).
- * To achieve large-scale production of bioactive recombinant bovine IL-12 (rboIL-12) using a baculovirus expression system.
Main Methods:
- * Construction of recombinant baculoviruses expressing p35 and p40 subunits of bovine IL-12.
- * Infection of insect cells and subsequent analysis of protein expression (monomers, dimers) via immunoblotting.
- * Superinfection strategy to facilitate heterodimer formation and functional assessment.
Main Results:
- * Recombinant baculovirus expressing the p40 subunit produced 40kDa monomers and 80kDa homodimers.
- * The p35 subunit was expressed as a 30kDa monomer in infected cells.
- * Superinfection yielded a 70kDa disulfide-bonded heterodimer in the supernatant, which induced IFN-gamma mRNA synthesis and production in bovine peripheral blood mononuclear cells.
Conclusions:
- * A baculovirus expression system can efficiently produce functional recombinant bovine IL-12.
- * The developed system allows for large-scale production of bioactive rboIL-12, suitable for further immunological studies and potential applications.