Construction of a eukaryotic vector containing human TACI-linker-BR3-IgGFc double receptor fusion gene and its expression in COS-7 cells
- VernacularTitle:人TACI-linker-BR3双受体融合基因真核表达载体构建及其在COS-7细胞中的表达
- Author:
Yan WANG
- Publication Type:Journal Article
- Keywords:
BAFF;
APRIL;
TACI;
BR3;
fusion protein;
eukaryotic expression
- From:
Academic Journal of Second Military Medical University
2000;0(11):-
- 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 bioactive,which paves a way for further research on the pathogenesis of autoimmune diseases and biotherapy strategy.
