Construction of Gadd45a expression plasmid and its expression in human T cells
10.3969/j.issn.1672-7347.2011.05.005
- VernacularTitle:Gadd45a表达质粒的构建及在人类T细胞中的表达
- Author:
Yaping LI
;
Ming ZHAO
;
Qianjin LU
- Publication Type:Journal Article
- Keywords:
Gadd45a;
pcDNA3.1;
Jurkat cells;
CD4+T cells
- From:
Journal of Central South University(Medical Sciences)
2011;36(5):398-404
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct Gadd45a expression plasmid and induce its expression in human T cells.Methods Gadd45a was amplified by reverse transcription PCR from human embryonic stem cells,and cloned into the pcDNA3.1 vector.The recombinant plasmid or blank plasmid was transfected into Jurkat cells or normal human CD4+T cells using electroporation,and the expression of Gadd45a was detected by quantitative RT-PCR and Western blot.Results Human Gadd45a expression plasmid was constructed successfully.Gadd45a was overexpressed both in Jurkat cells and normal human CD4+T cells after these cells were transfected with pcDNA3.1-Gadd45a.Conclusion The construction of Gadd45a expression plasmid and induction of Gadd45a overexpression in human T cells lay the foundation for further research on the role of Gadd45a in the epigenetic mechanism.
