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Free keywords:
gene regulation; lacZ; noncoding RNAs; translational fusion; mini-Tn7;
Pseudomonas
Abstract:
We describe the construction and validation of five mini-Tn7 vectors for analysis of post-transcriptional gene expression
in Pseudomonas. Four vectors allowconstruction 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 Ptac or PdctA. 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