Recombinant Eg-Grx1 Protein Expression

The full coding sequence of Eg-Grx1 was amplified from E. granulosus cDNA using primers 5ʹ-CCG GAA TTC ATG TGG CGC TTT TTA TC-3ʹ and 5ʹ-CCG CTC GAG CTC TAA AAG TTC AGC AAG TG-3ʹ, and then integrated into the EcoRI/XhoI restriction sites of vector pET28a (Novagen, Heidelberg, Germany). Recombinant protein was expressed and purified as previously described [30 (link)]. Briefly, the plasmid was transformed into Escherichia coli BL21 (DE3) cells (Cowin Biotech, Beijing, China) and induced with 0.8 mM isopropyl-1-thio-β-D-galactopyranoside at 37 °C for 6 h. Proteins were then purified using a Ni²⁺ affinity column (Bio-Rad, Hercules, CA) following the manufacturer’s instructions. The purified rEg-Grx1 protein was monitored by 15 % SDS-PAGE, and the protein concentration was estimated with a bicinchoninic acid protein assay kit (Pierce, Rockford, IL).