Ptrchis ta vector

His-tag protein expression vectors were constructed using pTrcHis TA vector (Themo Fisher Scientific) according to the manufacturer’s instructions as previously described [3 (link)]. In brief, PCR amplified target genes were TA-cloned into the pTrcHis TA vector (Themo Fisher Scientific) and inserted DNA was verified by sequencing (Genewiz, South Plainfield, NJ, USA). Proteins were expressed in TOP10 competent cells (Themo Fisher Scientific) with IPTG (Themo Fisher Scientific) at 1 mM for 2.5 h at 37 °C, and were purified using a Nuvia IMAC Nickel-charged column (Bio-Rad) on an NGC Quest chromatography system (Bio-Rad). Proteins underwent dialysis against PBS using Slide-A-Lyzer Dialysis Cassettes, 10 K MWCO (Themo Fisher Scientific).