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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 .
In E. coli BL21(DE3) the expression of the T7-RNAP is suppressed by the constitutively expressed LacI repressor. LacI binds the lac operator, which is located downstream of the LacUV5 promoter, preventing the production of the T7-RNAP. However, upon supplementation of IPTG, the LacI repressor dissociates from the lac operator, allowing for the ...
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]
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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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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 ...