Mobile website
Basic Research
Expression of mSDF-1γ/GM-CSF fusion protein and its protective effect on radiotherapyinduced hematopoiesis damages
- Article
- Figures
- Metrics
- Preview PDF
- Reference
- Related
- Cited by
- Materials
Abstract:
Objective: To express mSDF1γ/GMCS fusion protein in the Pichia pastoris expression system and investigate its in vivo promoting effects on hematopoiesis function and immune function. Methods: The mSDF1γ/GMCS fusion gene was chemically synthesized and was cloned into pichia expression vector; the vector was then transfected into Pichia pastoris expression system. Then the expressed products were detected by SDSPAGE，Western blotting and purified by ion exchange columns．Rat irradiation model was established by 60Co γ ray and were subcutaneously injected with the fusion protein. The hematopoietic and chemotactic activities of the fusion protein were investigated in vivo. Results: The expression vector pPIC9KSDF1LGMCSF was successfully constructed, and mSDF1γ/GMCS fusion protein was successfully expressed by Pichia pastoris strain GSl15．The molecular weight of the protein was about 32000 D, with a concentration of 78 ng/ml. The numbers of bone marrow mononuclear cells and GMCFU were significantly increased in rats after subcutaneous injection of the fusion protein (P<0.01). The apoptotic rate of bone marrow cells was significantly decreased（P<0.05 or P<0.01）, and the numbers of CD3+CD4+T lymphocytes in the peripheral blood were significantly increased (P<0.05). Conclusion: The mSDF1γ/GMCS fusion protein can improve the hematopoietic function and immune function in irradiated rats, which might be a promising cytokine for regulating hematopoiesis and antitumor immune response．
Keywords:
Project Supported:
Supported by the Natural Science Foundation of Shanghai（No. 06ZR14114）
Clc Number:
