Cloning vectors for gene delivery, integration and expression in Campylobacter jejuni

Abstract

Campylobacter genetics research is negatively impacted by a shortage of molecular tools for expressing DNA elements. A previous technique coupled an antibiotic resistance gene and its promoter to a gene of interest, inserting this expression unit into a conserved chromosomal location. Here the authors describe two new plasmids for construction and gene integration utilizing aspects of the previous type of expression unit. pBlueKan+cysMPro allows for the assembly of amplified DNA targets behind a kanamycin resistance marker and a constitutively transcribed cysM promoter. Transfer of the transcription unit to plasmid pCJR01 adds flanking regions of Campylobacter rRNA homology for recombination into conserved rRNA regions. System utility was demonstrated by restoring function of a flaAB deletion (RM3194ΔflaAB::tet) with a flaA gene or flaA/flaB combination.

Keywords

• Campylobacter
• fimbriae
• gene expression
• plasmids
• restored function