Cloning and sequencing of the genes for N-acetylglucosamine use that construct divergent operons (nagE-nagAC) from Vibrio cholerae non-O1.

Abstract

A 7.2-kb genomic DNA fragment containing N-acetylglucosamine-6-phosphate deacetylase gene (nagA) was cloned from the chitinase-producing bacterium Vibrio cholerae non-O1 strain 1148A (IFO 15429). Sequence analysis of the DNA fragment found three other complete open reading frames (ORFs) and the 5' end of an ORF. Amino acid sequences of two ORFs, ORF2 and ORF4, showed similarity with that of NagC, the repressor of nag operons and that of NagE, N-acetylglucosamine-specific transporter II(Nag) of phosphoenolpyruvate transport system of Escherichia coli, respectively. In the presence of N-acetylglucosamine, nagA and ORF2 (nagC) were co-transcribed. ORF4 (nagE), which is upstream from nagAC but is expressed in the opposite direction was also transcriptionally induced in the presence of N-acetylglucosamine. These results indicated that nagE-nagAC existed as divergent operons in V. cholerae non-O1. Unlike E. coli, nagB and nagD were not in the operons.