In-frame deletion of Escherichia coli essential genes in complex regulon

A conditional knockout-rescue system was developed to construct an in-frame deletion strain of Escherichia coli essential genes. The target was flanked with marker genes and FRT (FLP recognition target) sites, and a plasmid containing arabinose-induced FLP recombinase was transformed. After arabinose induction, cells could survive only when target protein activity was provided in trans. We selected three essential genes as targets, yaeT, fabZ, and dnaE, which are components of the complex eight-gene regulon yaeT-hlpA-lpxD-fabZ-lpxA-lpxB-rnhB-dnaE. Deletion of these three genes exhibit no polar effects on their adjacent genes in terms of cell viability, meaning that this system not only allows for the simplified study of protein interactions and homolog screening in other organisms, but also facilitates the null mutant construction of essential genes.