Expression and purification of GRIK3 intracellular region protein in prokaryotic cell system / 实用医学杂志

ABSTRACT

Objective To express and purify GRIK3 intracellular region protein in prokaryotic cell system. Methods The histamine(His)tagged GRIK3 intracellular region recombinant plasmid pCzn1-GRIK3-in-tra was constructed and transformed into Escherichia coli(E.coli). The His-GRIK3-intra recombinant protein was expressed after the induction with IPTG. The target protein was purified by Ni-NTA affinity chromatography column. Results The prokaryotic expression plasmid pCzn1-GRIK3- intra was successfully constructed and the GRIK3 intracellular region protein in E.coli were efficiently expressed after the induction with IPTG. The purified target pro-tein was successfully obtained by Ni-NTA affinity chromatography column. Conclusion The successful construc-tion of prokaryotic expression plasmid expressing GRIK3 intracellular region gene and preparation of high purity GRIK3 intracellular region protein have paved the way for further exploration of the intracellular signal transduction mechanism of membrane protein GRIK3.