The corresponding BL21 DE3) and BL21 strains were grown at 37 °C overnight in Luria-Bertani (LB) medium containing ampicillin.
Cells were grown at 37 °C in 1 L of LB medium containing ampicillin and kanamycin until an A600 = 0.6 0.8 was reached.
The cultures were diluted 1 100 with auto-inducible ZYP5052 medium containing ampicillin and subjected to further incubation at 20 °C until the culture density reached saturation.
Transformed E. coli BL21 (DE3) cells were incubated with medium containing ampicillin for 3 h at room temperature, and then IPTG (Sigma-Aldrich) was added to a final concentration of 100 μM.
For the Escherichia coli expression culture 100 ml LB medium containing ampicillin (100 µg/ml) and chloramphenicol (25 µg/ml) was inoculated with 5 ml of an overnight culture of the Rosetta (DE3) pLysS strain (Novagen) previously transformed with pMG307 and grown at 37 °C until an OD600 of 0.6 was reached.
A 1 % culture of the bacterial solution in LB medium containing ampicillin and kanamycin was prepared.
Luria-Bertani medium containing ampicillin (100 μg/ml) was used to cultivate E. coli.
Transformants were grown in 1200 mL of Luria-Bertani medium containing ampicillin (100 μg/mL) and kanamycin (50 μg/mL).
Then, 5 mL of the overnight cultures were transferred into 1 L fresh LB medium containing ampicillin in 2 L flask.
White clones were picked randomly and inoculated into 1 mL of Luria Broth (LB) medium containing ampicillin and incubated at 37°C for 24 h.
Escherichia coli DH5a cells were transformed with pUC19 plasmid DNA and then grown overnight in the LB medium containing ampicillin (50 μg/ml) at 37°C.
