Identification of Campylobacter jejuni Promoter Sequences

Abstract

ABSTRACT A promoterless lacZ shuttle vector, which allowed screening of promoters by β-galactosidase activity in Campylobacter jejuni and Escherichia coli , was developed. Chromosomal DNA fragments from C. jejuni were cloned into this vector; 125 of 1,824 clones displayed promoter activity in C. jejuni . Eleven clones with strong promoter activity in C. jejuni were further characterized. Their nucleotide sequences were determined, and the transcriptional start sites of the putative promoters in C. jejuni were determined by primer extension. Only 6 of these 11 promoters were functional in E. coli . The 11 newly characterized and 10 previously characterized C. jejuni promoters were used to establish a consensus sequence for C. jejuni promoters. The 21 promoters were found to be very similar. They contain three conserved regions, located approximately 10, 16, and 35 bp upstream of the transcriptional start point. The −10 region resembles that of a typical ς 70 E. coli promoter, but the −35 region is completely different. In addition a −16 region typical for gram-positive bacteria was identified.