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Published on: May 10, 2017
[Cloning and expression of human Runx3 gene obtained from human peripheral blood CD8(+) T lymphocyte]
Ya-zhen Li1, Sheng-jun Wang, Yan Shi
1Department of Immunology, Institute of Clinical Laboratory Medicine, Jiangsu University, Zhenjiang, China. lyz-0115@163.com
Aim:
Runx3, a type of Runt family member, plays an important role in immune regulation. In this study, we cloned and analyzed the cDNA encoding human Runx3 from T lymphocyte, expressed Runx3 protein in E.coli system, and studied the relation between Runx3 and some immune disorders or tumors.
Methods:
The CD8(+) T were isolated from human peripheral blood with MACS, Runx3 cDNA was amplified by RT-PCR and cloned into pMD19-T vector, and recombinant was transformed into competent cells DH5alpha and recombinant sequencing were performed. The identical was subcloned into pQE30 vector and expressed in E.coli M15. The fusion protein was identified by Western blot.
Results:
The 1,248 bp fragment amplified by RT-PCR was the same as the anticipated one in size and encodes 415 amino acids. Runx3 protein was gained.
Conclusion:
Human Runx3 gene was cloned and expressed in E.coli system successfully, which brought a foundation for further research on its biological function.
