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Isopropyl β-d-1-thiogalactopyranoside (IPTG) is a molecular biology reagent. This compound is a molecular mimic of allolactose , a lactose metabolite that triggers transcription of the lac operon , and it is therefore used to induce protein expression where the gene is under the control of the lac operator .
While E. coli BL21(DE3) supports the expression of genes under the control of constitutive promoters, it is specifically engineered for IPTG induction of recombinant genes under the control of a T7 promoter. The realized induction strength depends on several factors, including the IPTG concentration and the timing of its supplementation. [5]
IPTG is a reagent which mimics the structure of allolactose, and can therefore bind to the lac repressor and prevent it from inhibiting gene expression. Once enough IPTG is added, the T7 gene is normally transcribed and so transcription of the gene of interest downstream of the T7 promoter also begins. [6]
Beta-gal is inhibited by L-ribose and by competitive inhibitors 2-phenylethyl 1-thio-β-D-galactopyranoside (PETG), D-galactonolactone, isopropyl thio-β-D-galactoside (IPTG), and galactose. [ 4 ] β-Galactosidase is important for organisms as it is a key provider in the production of energy and a source of carbons through the break down of ...
IPTG, a molecule similar to lactose, but with a sulfur bond that is not hydrolyzable so that E. coli does not digest it, is used to activate or "induce" the production of the new protein. Once the cells are induced, it is difficult to remove IPTG from the cells and therefore it is difficult to stop expression.
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