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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 .
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]
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]
The lactose operon (lac operon) is an operon required for the transport and metabolism of lactose in E. coli and many other enteric bacteria.Although glucose is the preferred carbon source for most enteric bacteria, the lac operon allows for the effective digestion of lactose when glucose is not available through the activity of β-galactosidase. [1]
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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Its production may be induced by a non-hydrolyzable analog of allolactose, IPTG, which binds and releases the lac repressor from the lac operator, thereby allowing the initiation of transcription to proceed. It is commonly used in molecular biology as a reporter marker to monitor gene expression.