Mini-Tn7 vectors for studying post-transcriptional gene expression in Pseudomonas

We describe the construction and validation of five mini-Tn7 vectors for analysis of post-transcriptional gene expression in Pseudomonas. Four vectors allow construction of translational fusions to β-galactosidase (lacZ), while the fifth is designed for functional analysis of noncoding RNA genes. Translational fusions can be constructed without a functional promoter in the vector or from an inducible promoter of either P(tac) or P(dctA). We show that promoterless fusions have value for determining levels of translation, whereas fusions to inducible promoters have utility in the analysis of mRNA-binding factors.