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Updated: Jul 19, 2026
Proteomics to Identify Proteins Interacting with P2X2 Ligand-Gated Cation Channels
Published on: May 18, 2009
[Construction of pGEX4T-1-Cox7a2 and expression, purification and identification of the recombinant protein]
Liang Chen1, Zhong-cheng Xin, Xue-jun Jiang
1Andrology Center, the First Hospital of Peking University, Beijing 100009, China. bdyychenliang@163.com
Objective:
To clone and express Cox7a2, one mitochondrial respiratory chain related gene, and to identify its recombinant protein.
Methods:
The coding region of Cox7a2 was amplified from primary cultured mouse Leydig cells by RT-PCR. The PCR product was cloned into pGEX4T-1 vector by BamH I and EcoR I sites, and confirmed by DNA sequencing. The recombinant fusion protein vector was transformed and expressed into BL21. The recombinant fusion protein was identified by Western blotting.
Results:
The entire coding region of Cox7a2 was cloned and expressed. The fusion protein was identified by anti-GST monoclonal antibody using Western blotting.
Conclusion:
The cloning of Cox7a2 and the expression of the recombinant protein would help to study the detailed function of Cox7a2, one respiratory chain related and highly differently expressed gene in the tissues of aging testes.
