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Recombinant OmpA22-350 (rOmpA) protein was expressed with hexahistidine affinity tags at their N termini using the expression vector TAGZyme pQE2 (Qiagen Sciences, Germantown, MD). The gene fragment was amplified using PCR and Pfu polymerase (Promega KK, Tokyo, Japan), E. coli strain ATCC 25922T genomic DNA as the template, and the primers Fw_rOmpA22 and Rv_rOmpA350. The PCR fragments were cloned into pQE2 at the SalI and XhoI sites, and the resulting plasmids were transformed into E. coli BL21 (DE3) (GE Healthcare, Tokyo, Japan) for protein expression. The recombinant proteins were purified using Ni2 + -chelate chromatography. |
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using PCR and Pfu polymerase (Promega KK, Tokyo, Japan), E. coli strain ATCC 25922T genomic DNA as the template, and the primers Fw_rOmpA22 and Rv_rOmpA350. The PCR fragments were cloned into pQE2 at the SalI and XhoI sites, and the resulting plasmids were transformed into E. coli BL21 (DE3) (GE Healthcare, Tokyo, Japan) for protein expression. The recombinant proteins were purified using Ni2 + -chelate chromatography.
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