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2.1. Protein expression rBoNT/A was produced by Claruvis Pharmaceuticals (Chongqing). The gene encoding the amino acids of BoNT/A1 was cloned into pGex vector with a GST tag introduced at the N-terminus. Two peptide linkers containing an HRV-3C protease cleavage site were inserted between the LC and HC and between the GST-tag and LC. The recombinant protein was expressed in E. coli BL21 Star (DE3) cells (One Shot, Invitrogen).
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