pET-28a(+)-g-α13

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	The recombinant plasmids were transformed into E. coli BL21(DE3) (TransGen Biotech, Beijing, China). Freshly transformed bacteria were inoculated into LB medium (50 μg/mL kanamycin) and grew at 37°C with continuous shaking cultivation at 200 rpm until the absorbance at 600 nm reached 0.4~0.6. The α-13 giardin fusion protein was expressed under the induction of isopropyl-1-thio-β-D-galactopyranoside (IPTG).

Protocol tips

The recombinant plasmids were transformed into E. coli BL21(DE3) (TransGen Biotech, Beijing, China). Freshly transformed bacteria were inoculated into LB medium (50 μg/mL kanamycin) and grew at 37°C with continuous shaking cultivation at 200 rpm until the absorbance at 600 nm reached 0.4~0.6. The α-13 giardin fusion protein was expressed under the induction of isopropyl-1-thio-β-D-galactopyranoside (IPTG).

Publication protocol

The recombinant plasmids were transformed into E. coli BL21(DE3) (TransGen Biotech, Beijing, China). Freshly transformed bacteria were inoculated into LB medium (50 μg/mL kanamycin) and grew at 37°C with continuous shaking cultivation at 200 rpm until the absorbance at 600 nm reached 0.4~0.6. The α-13 giardin fusion protein was expressed under the induction of isopropyl-1-thio-β-D-galactopyranoside (IPTG); firstly, the expression conditions were optimized by adjusting IPTG concentrations (0.2, 0.4, 0.6, 0.8, 1.0, and 1.2 mM), induction time (1, 3, 5, and 7 h), and culturing temperature (16, 20, 25, 30, and 37°C). Then, they were expressed by autoinduction expression system (ZYM-5052); the bacterium solutions cultivated by ZYM-5052 for 3~16 h were collected, respectively. For details, refer to Studier [15] with few modifications. The cultures were harvested at 9600 ×g for 10 min at 4°C after washing the cell pellet in phosphate buffer (containing 0.1% Tween-20).

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