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Preparation of SDF-1γ/rhGM-CSF fusion protein and its chemotactic effect
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Abstract:
Objective : To prepare the fusion protein of SDF-1 and rhGM-CSF (SDF-1γ/rhGM-CSF) by genetic engineering technology, and investigate its hematopoietic and immune promotion functions in tumor patients. Methods: The expression vector for SDF-1γ/rhGM-CSF fusion protein, pPIC9k-SDF1-rhGM-CSF1, was constructed and the protein expression was induced by yeast transfection. SDF-1γ/rhGM-CSF fusion protein was further identified by Western blotting analysis. Colony-formation assay and chemoattract assay were used to study the roles of the prepared fusion protein in stimulating bone marrow cell colony-formation and in chemoattracting immature dendritic cells. Results: SDF-1γ/rhGM-CSF fusion gene vector, pPIC9k-SDF1-rhGM-CSF1, was successfully constructed and expressed high level of SDF-1γ/rhGM-CSF fusion gene. The molecular weight of the expressed protein was about 25 000 and was recognized by GM-CSF specific antibody. The fusion protein had a stronger effect in stimulating bone marrow cell colony-formation than GM-CSF (P<005) and in chemoattracting immature dendritic cells than SDF-1 (P<0.05). Conclusion: SDF-1γ/rhGM-CSF fusion protein can promote bone marrow cell colony-formation and chemoattraction of immature dendritic cells, which might be used for promoting hematopoiesis and immune function of tumor patients after chemotherapy.
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Project Supported:
Project supported by the Natural Science Foundation of Shanghai (No.06ZR14114), and the "11th Five-Year Plan" Program of PLA (No.10MA004)
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