Construction of a eukaryotic vector containing human TACI-linker-BR3-IgGFc double receptor fusion gene and its expression in COS-7 cells
- Author:
Yan WANG
1
Author Information
1. Department of Clinical Diagnosis
- Publication Type:Journal Article
- From:
Academic Journal of Second Military Medical University
2006;27(11):1190-1195
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To construct a eukaryotic vector containing human TACI-linker-BR3-IgGFc double receptor fusion gene and to investigate its expression in COS-7 cells. Methods: The standard cloning technology was employed to construct the title vector, which was then used to transfect COS-7 cells by lipofectamine 2000. The expression of target protein and their interaction with BAFF and APRIL protein were examined by Western blot, ELISA, and immunohistochemistry methods. Results: The expression of the fusion protein was observed in transiently transfected COS-7 cells, with the transfection efficiency being 48%. The fusion protein was detected in the supernatant. It suggested that the fusion protein interacted well with BAFF and APRIL proteins; the IgGFc tag did not influence the natural assembly and the biological activity of fusion proteins. Conclusion: A vector containing TACI-linker-BR3 double receptor and IgGFc fusion gene has been successfully constructed and the subsequent fusion protein is bloactive, which paves a way for further research on the pathogenesis of autoimmune diseases and biotherapy strategy.
