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Construction of Gadd45a expression plasmid and its expression in human T cells
Yaping Li1, Ming Zhao, Qianjin Lu
1Department of Dermatology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
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.
