Single colonies of E.coli BL21 (DE3) and E.coli Rosetta (DE3) transformed with the recombinant expression plasmid pET28a-VP2 were respectively inoculated in the tube containing 5 ml of LB medium (containing 50 µg/mL of kanamycin, 50 µg/mL of kanamycin and 34 µg/mL of chloramphenicol) and cultivated overnight at 37 °C in a shaking incubator. Until the OD600 of the culture was about 0.6–0.8, the expression of VP2 was induced by the addition of isopropyl β-D-thiogalactoside (IPTG). After induction, the expression of the fusion protein was analyzed by SDS-PAGE, and the cells were harvested at 25 °C by centrifugation at 10,000 rpm for 15 min. The supernatant was discarded and the cell pellet was washed, frozen, resuspended in phosphate-buffered saline (PBS, pH 7.4), and then disrupted by sonication. The eluted fractions were analyzed by 10% SDS-PAGE. The results of SDS-PAGE were obtained using a Universal Hood III gel imaging system (Biorad, US).

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