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Escherichia coli BL21(DE3) is a commonly used protein production strain of the E. coli bacterium. This strain combines several features that allow for excessive expression of heterologous proteins . It is derived from the B lineage of E. coli .
The cells are competent and often used with calcium chloride transformation to insert the desired plasmid. A study of four transformation methods and six bacteria strains showed that the most efficient one was the DH5 strain with the Hanahan method. [2]
In 1885, Theodor Escherich, a German pediatrician, first discovered this species in the feces of healthy individuals and called it Bacterium coli commune because it is found in the colon and early classifications of Prokaryotes placed these in a handful of genera based on their shape and motility (at that time Ernst Haeckel's classification of Bacteria in the kingdom Monera was in place [3]).
Cells are typically rod-shaped, and are about 2.0 μm long and 0.25–1.0 μm in diameter, with a cell volume of 0.6–0.7 μm 3. [19] [20] [21] E. coli stains gram-negative because its cell wall is composed of a thin peptidoglycan layer and an outer membrane.
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Typically, a sterile, filtered 1 mol/L solution of IPTG is added 1:1000 to an exponentially growing bacterial culture, to give a final concentration of 1 mmol/L. The concentration used depends on the strength of induction required, as well as the genotype of cells or plasmid used.
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