The vector is a suicide mini-transposon delivery plasmid and R6K based, being unable to survive in recipients lacking the Pir protein. Selection for the kanamycin or gentamycin resistance encoded by the mini-transposons yields recipients in which the mini-transposon, but not the cognate transposase gene present on the suicide plasmid, has integrated into one of the endogenous replicons. The modified GFP exhibits a 45-fold-increased fluorescent signal compared with that of the natural GFP. The enhanced sensitivity of this vector, gfp-based promoter-probe mini-transposon, allows the direct detection of transconjugants harbouring monocopy transcriptional fusions by means of a UV handy-lamp at 366 nm by the naked eye and the detection of single gfp-tagged bacterial cells by epifluorescence microscopy. Restriction digests of the clone give the following sizes (kb): NotI--5.7; ClaI--5.7; KpnI--4.1, 1.6. |
