CONSTRUCTION OF A TRANSLATIONAL LACZ FUSION SYSTEM TO STUDY GENE-REGULATION IN NEISSERIA-GONORRHOEAE

A translational lacZ reporter system to study gene regulation in Neiss eria gonorrhoeae (Ng) was developed. The pUC18-based vector pLES94 tra nsforms Ng and recombines into the Ng chromosome at the site of the pr oAB genes. The vector contains a restriction site for cloning promoter s that will result in a lacZ gene fusion. Initial cloning and characte rization of promoters can be done in Escherichia coli. The vector cont ains both Ap(R) and Cm-R genes, however the Ap(R) gene is lost when th e insert combines into the Ng chromosome. This system gives single cop y expression of the fusion and does not result in the inactivation of the gene of interest.