Expression, Purification and Characterization of Amantadine Receptor in Escherichia coli

Abstract:

In order to obtain large quantities of broadly selective receptor as one diagnose agent to detect amantadine residue, the M2 protein gene with a His-tag was ligated into pET11a and transferred into E. coli BL21 (DE3) cell. The recombinant E. coli was cultured in liquid LB culture. SDS-PAGE result showed the recombinant M2 protein (rM2) was expressed as insoluble inclusion bodies with about 18KDa in molecular size. rM2 protein was further recognized by Western blot and purified by Ni Sepharose 6 Fast Flow and then refolded. The equilibrium dialysis result showed the rM2 protein had the binding constant of 1.1×10⁵, and stoichiometry of 4.2. The above result showed the rM2 has the potential as biological diagnose agent to the detection of amantadine residue.

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