Expression of bioactive recombinant bovine interferon-gamma using baculovirus

Area of Science:

• Immunology
• Molecular Biology
• Biotechnology

Background:

• The role of bovine interferon-gamma (BoIFN-gamma) in disease and therapy is not well understood.
• BoIFN-gamma is implicated in immune defense against pathogens and potentially tumors.
• A functional recombinant form is needed to study its therapeutic potential.

Purpose of the Study:

• To express a fully functional recombinant bovine interferon-gamma (rBoIFN-gamma) using a baculovirus expression system.
• To characterize the recombinant protein and assess its biological activity.
• To enable further research into BoIFN-gamma as a therapeutic agent or vaccine adjuvant.

Main Methods:

• Bovine interferon-gamma cDNA was amplified from peripheral blood mononuclear cell (PBMC) RNA via reverse transcription-polymerase chain reaction (RT-PCR).
• The open reading frame (ORF) was cloned into the baculovirus transfer vector pBlueBac 4.5/V5 His.
• Recombinant baculovirus (AcBoIFN-gamma) was generated and used to infect Spodoptera frugiperda (Sf9) cells for protein expression.

Main Results:

• Recombinant BoIFN-gamma (rBoIFN-gamma His) was successfully expressed and secreted into the culture medium.
• Purified rBoIFN-gamma His was confirmed as a glycosylated protein of 20-22 KDa by SDS-PAGE and glycan analysis.
• The recombinant protein demonstrated in vitro antiviral activity against bovine viral diarrhoea-mucosal disease virus (BVD/MD).

Conclusions:

• The baculovirus system provides a method for producing bioactive rBoIFN-gamma His.
• This recombinant protein can be utilized for further investigation into its therapeutic applications in bovine diseases.
• rBoIFN-gamma His holds potential as a vaccine adjuvant or treatment for various bovine conditions.