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The recombinant plasmid OmpA-TRAIL/pET22b was transformed into the E.coli BL21 (DE3) by treatment with 0.1 M cold CaCl2 and then heat shock at 42 °C for 1 minute. The transformed bacteria was cultured in Terrific Broth (TB) medium (1.2% peptone; 2.4% yeast extract; 72 mM K2HPO4; 17 mM KH2PO4; and 0.4% glycerol; pH 7.2) on a rotary shaker (200 rpm) at 37 °C until the optical density at 600 nm (OD600) of the culture reached 0.6. Then, isopropyl-β-D-thiogalacto-pyran-oside (IPTG) was added to a final concentration of 0.1 mM to induce the expression of the target protein until 24 h. |
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Protocol tips |
The recombinant plasmid OmpA-TRAIL/pET22b was transformed into the E.coli BL21 (DE3) by treatment with 0.1 M cold CaCl2 and then heat shock at 42 °C for 1 minute. The transformed bacteria was cultured in Terrific Broth (TB) medium (1.2% peptone; 2.4% yeast extract; 72 mM K2HPO4; 17 mM KH2PO4; and 0.4% glycerol; pH 7.2) on a rotary shaker (200 rpm) at 37 °C until the optical density at 600 nm (OD600) of the culture reached 0.6. Then, isopropyl-β-D-thiogalacto-pyran-oside (IPTG) was added to a final concentration of 0.1 mM to induce the expression of the target protein until 24 h. |
Publication protocol
The recombinant plasmid OmpA-TRAIL/pET22b was transformed into the E.coli BL21 (DE3) by treatment with 0.1 M cold CaCl2 and then heat shock at 42 °C for 1 minute. The transformed bacteria was cultured in Terrific Broth (TB) medium (1.2% peptone; 2.4% yeast extract; 72 mM K2HPO4; 17 mM KH2PO4; and 0.4% glycerol; pH 7.2) on a rotary shaker (200 rpm) at 37 °C until the optical density at 600 nm (OD600) of the culture reached 0.6. Then, isopropyl-β-D-thiogalacto-pyran-oside (IPTG) was added to a final concentration of 0.1 mM to induce the expression of the target protein until 24 h.
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