An erythromycin resistance cassette and mini-transposon for constructing transcriptional fusions to cat

A new cassette (Er-Cm cassette) and mini-transposon (mTn) (Tn MaxErCm) based on the previously described mTn, Tn Max2 [Haas et al., Gene 130, 23–31.], have been constructed. The cassette and mTn make use of an erythromycin resistance (Er R) marker encoded by ermC′. Both the Er-Cm cassette and Tn MaxErCm also carry a promoterless cat gene to allow the construction of transcriptional fusions and the measurement of transcriptional activity by assaying for chloramphenicol acetyltransferase. We show the function of these genetic elements by analyzing the regulation of expression of the mglA gene of Francisella novicida and by using Tn MaxErCm to probe for promoter activity within an F. novicida recombinant clone. The reporter cassette and mTn described here further expand the family of Tn Max transposons and facilitate the study of gene expression in organisms where direct Tn mutagenesis methods are unavailable.