pTrcHis-TOPO

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	The plasmids were transformed into BL21 (DE3) competent E. coli (New England BioLabs) using a standard protocol. Transformed bacterial cells were grown in Luria Bertani broth at 37°C with constant shaking until reaching optical density of A600=0.6. Protein production was induced in the cultured cells with the addition of 0.1 mM isopropyl-1-thio-β-d-galactopyranoside and 5 μM FeSO4. The cells were allowed to grow overnight at 16°C (human BCO1) or for 8 hours at 16°C (murine BCO2).

Protocol tips

The plasmids were transformed into BL21 (DE3) competent E. coli (New England BioLabs) using a standard protocol. Transformed bacterial cells were grown in Luria Bertani broth at 37°C with constant shaking until reaching optical density of A600=0.6. Protein production was induced in the cultured cells with the addition of 0.1 mM isopropyl-1-thio-β-d-galactopyranoside and 5 μM FeSO4. The cells were allowed to grow overnight at 16°C (human BCO1) or for 8 hours at 16°C (murine BCO2).

Publication protocol

The plasmids were transformed into BL21 (DE3) competent E. coli (New England BioLabs) using a standard protocol. Transformed bacterial cells were grown in Luria Bertani broth at 37°C with constant shaking until reaching optical density of A600=0.6. Protein production was induced in the cultured cells with the addition of 0.1 mM isopropyl-1-thio-β-d-galactopyranoside and 5 μM FeSO4. The cells were allowed to grow overnight at 16°C (human BCO1) or for 8 hours at 16°C (murine BCO2).

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