Molecular cloning of chemotaxis genes and overproduction of gene products in the bacterial sensing system

Abstract

The chemotaxis genes cheR, cheB, cheY, cheZ, and tar of Salmonella typhimurium were cloned into bacteriophage lambda vectors and onto pBR322 plasmids by recombinant DNA techniques. The genes were linearly arranged in the order tar-cheR-cheB-cheY-cheZ (and were read from a promoter on the upstream side of the tar or cheR gene). However, their stoichiometries of expression were found to be 4:1:1:18:3, respectively. The overexpression of the cheY gene appeared to be a function of translational control. These five che genes were placed on a multicopy plasmid, and the gene products were overproduced in the cells, as shown by enzyme assays. The overproduction of the products of these five genes relative to those of the other che genes caused some changes in chemotactic properties, but no dramatic destruction of sensing ability.