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. Author manuscript; available in PMC: 2020 Jul 30.
Published in final edited form as: J Vis Exp. 2019 Jun 20;(148):10.3791/59729. doi: 10.3791/59729

Figure 1. Amplification of competent E. coli and induction of protein expression.

Figure 1

(A) Insertion of the pET vector into competent BL21(DE3) pLysS E. coli bacteria, described in section 1. (B) Heat shock protocol and plating of the pET transformed E. coli bacteria, described in step 1 of the protocol. Transformed bacteria are plated on LB agar plates with antibiotics for selection. (C) Amplification of pET transformed E. coli bacteria, described in section 1. Colonies are picked from an LB agar plate and amplified in nourishing medium (LB or NZCYM) until the bacterial density reached the empirical threshold of 0.4. (D) Induction of protein expression via the DE3-IPTG-pET system, described in section 1 and sketched in Figure 2. Protein production is started by the application of the chemical IPTG. At the end of section 1, the bacterial pellet containing the protein is harvested. Please click here to view a larger version of this figure.