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Expression of MIP-2γ/GM-CSF Fusion Protein and Preliminary Study of Its Biological Bifunction
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Abstract:
Objective: To generate a new kind of cytokine MIP-2γ/GM-CSF fusion protein by genetic engineering technology and investigate its biological bifunction. Methods: The eukaryotic expressing vector for MIP-2γ/GM-CSF fusion protein was constructed by primer design and step by step cloing. MIP-2γ gene and GM-CSF gene were linked via a linker sequence（Gly4Ser）3. After transient expression， the cell supernatants of 293-T cells containing the aimed protein were obtained. The hematopoietic and chemotactic activities of the fusion protein were investigated in vitro.Results: MIP-2γ/GM-CSF fusion gene vector was confirmed by restriction analysis and sequencing analysis. The molecular weight of the protein was about 24.9 kD. MIP-2γ/GM-CSF fusion protein could stimulate the proliferation of TF-1 cells like GM-CSF. The specific activity of the fusion protein was 2.2×107IU/mg. The fusion protein was found to be effective in stimulating bone marrow cell colony-formation and chemoattracting immature dendritic cells and CD3+T cells. Conclusion: MIP-2γ/GM-CSF fusion protein exhibits bifunctions which including the promotion of hematopoiesis and chemoattration of immune cells, suggesting that the fusion protein can be used to regulate hematopoiesis and antitumor immune response as a new kind of cytokine.
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