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Isopropyl-beta-D-thiogalactopyranoside, short as IPTG, is a very powerful inducer, stable in nature and not metabolized by bacteria, so it is widely used in laboratories. IPTG is often used in cloning experiments that require the induction of β-galactosidase activity. It is often used in combination with X-Gal or Bluo-Gal for blue-and-white screening of recombinant bacterial colonies, which can induce the expression of the lac operon in E. coli. IPTG binds to lacI repressor protein and changes its conformation to play a role, preventing the inhibition of β-galactosidase encoding gene lacZ.
1. Molecules that can induce the synthesis of enzymes but are not broken down are called comfort inducers. Although lactose can induce the synthesis of enzymes, it decomposes with it, resulting in many complicated kinetic problems. Therefore, people often use comfort inducers for various experiments.
2. IPTG is not metabolized by bacteria and is a lactose analogue. Once it enters the cell, it will have a long-lasting induction effect. Therefore, the induction efficiency of IPTG is high, and often only a small amount (1mmol/L) is needed to achieve the ideal induction effect. . Since IPTG is not metabolized, it specifically induces the expression of foreign proteins in the cell. Unaffected by cell metabolism, the induction effect remains stable. IPTG is suitable for experimental research due to its excellent stability.
3. IPTG can be copied effectively without lacY gene.
4. Sulfur replaces oxygen in the galactosidic bond and loses its hydrolytic activity. However, thiogalactoside and homologous oxo compounds have the same affinity for the enzyme site. Although IPTG is not controlled by β-galactosidase Recognition, but it is a very effective inducer of the lac gene cluster.
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