Introduction of IPTG [367-93-1]

IPTG induction principle(http://www.hnhbsj.com)
First, the lactose operon (element) of, E. coli contains three structural genes, ZHY and A, which encode galactosidase, osmotic enzyme and acetyltransferase, respectively. A priming sequence P and a regulatory gene I.I gene encode a repressor protein, which binds to the O sequence and causes the operon (element) to be repressed and closed. There is also a (CAP) binding site in the catabolic (metabolite) gene upstream of the priming sequence P. The regulatory region of lac operon is composed of P sequence, O sequence and CAP binding site. A regulatory region is regulated to achieve the coordinated expression of gene products.

Secondly, when lactose is not present, the lac operon (element) is in the repressor state. At this time, the Lac repressor protein expressed by the I sequence under the manipulation of the PI priming sequence binds to the O sequence, hinders the binding of the RNA polymerase to the P sequence and inhibits the transcriptional initiation. When lactose is present, the lac operon can be induced. In this operon system, the real inducer is not lactose itself. Lactose enters cells and is converted to isolactose catalyzed by β-galactosidase. The latter is a molecular binding repressor protein as an inducer, which changes the conformation of the protein, resulting in the dissociation of the repressor protein from the O sequence and the transformation of the repressor protein. Record. Isopropylthiogalactoside (IPTG) has the same effect as isolactose and is a powerful inducer, which is not metabolized by bacteria and is very stable, so it is widely used in laboratory.

usage method(http://www.hnhbsj.com)
Firstly, IPTG was prepared into 23.83 mg/ml (100 mM) aqueous solution. Then the IPTG,X-Gal,Amp plate medium was prepared by adding 100 μ l X-Gal (20 mg/ml dimethylformamide (DMF) solution) and 100 μ l Amp (100 mg/ml) into 100 ml Agar medium. When the DNA fragment is inserted into the pUC family of vectors (or any other vector with the lacZ,Amp gene), it is then transformed into the After the lacZ deletion cells were coated with the IPTG,X-Gal,Amp plate medium, the gene recombinant could be conveniently selected according to the blue and white growth of the cell (white is the gene recombinant with DNA insert fragment).

act on(http://www.hnhbsj.com)
The expression of exogenous gene is generally used in prokaryotic expression system, and the expression level is increased, the product is stable and easy to identify.

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