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[Expression of human interleukin-12 in Baculovirus expression system]
1Biopharmeutical Center, State Key Lab for Biocontrol, Zhongshan University, Guangzhou 510275, China.
Wei Sheng Wu Xue Bao = Acta Microbiologica Sinica
|January 29, 2003
Summary
Researchers successfully expressed and secreted human Interleukin-12 (hIL-12) p35 and p40 subunits in insect cells. The resulting proteins demonstrated immunogenicity, paving the way for further research into this key cytokine.
Area of Science:
- Immunology
- Molecular Biology
- Biotechnology
Context:
- Human Interleukin-12 (hIL-12) is a critical immunoregulatory cytokine.
- hIL-12 is unique as a disulfide-linked heterodimer, comprising p35 and p40 subunits.
Purpose:
- To construct recombinant transfer vectors for hIL-12 p35 and p40 genes.
- To express and secrete these subunits in insect cells using a baculovirus system.
- To confirm the successful expression and immunogenicity of the recombinant proteins.
Summary:
- Recombinant transfer vectors pAcAB3-p35 and pAcAB3-p40 were created.
- These vectors, along with linearized baculovirus DNA, were co-transfected into Sf9 insect cells.
- Recombinant viruses AcNPV-OCC(-)-hiL-12 (p35) and AcNPV-OCC(-)-hIL-12 (p40) were generated and used for infection.
- SDS-PAGE and Western blot analysis confirmed successful expression and secretion of both hIL-12 p35 and p40 subunits.
- The expressed proteins exhibited immunogenicity.
Impact:
- Demonstrates a viable method for producing immunogenic hIL-12 subunits in insect cells.
- Provides a foundation for further studies on hIL-12 function and therapeutic applications.
- Highlights the potential of baculovirus expression systems for cytokine production.