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Establishment of a low-dosage-IPTG Inducible Expression System Construction Method in Escherichia Coli

Establishment of a low-dosage-IPTG Inducible Expression System Construction Method in Escherichia Coli

Ming Zhao, Xin-Yi Tao, Feng-Qing Wang, Yu-Hong Ren, Dong-Zhi Wei

J Basic Microbiol. 2018 Sep;58(9):806-810.

PMID: 29962051

Abstract:

The lac operon is a delicate inducible gene expression element in bacteria. To efficiently induce gene expression, a sufficient dosage of an inducer, usually that of 500-1000 µM isopropyl β-D-1-thiogalactopyranoside (IPTG), is required to keep repressor LacI from its binding sites, which is a heavy cost burden in low-value-added products. So we propose a strategy to reduce the required dosage of IPTG by restricting LacI expression. To test this strategy, we employed a reconstructed IPTG inducible expression system based on lac operon, Promoter(lacO)-target gene-PtacL-lacI, where a modified promoter, Ptac, with a random synthetic library (PtacL) to instead of PlacI to optimize LacI expression in Escherichia coli. Finally, the PtacL mutant, PtacL4, which could maintain the same repression effect as the original PlacI while reducing the required dosage of IPTG from 500 to 20 µM, was selected. This method is simple and efficient and can be of a good reference point for attempts to reduce inducer concentration in the IPTG or similar inducible expression systems.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP367931-B	Isopropyl β-D-1-thiogalactopyranoside		367-93-1	Price

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