The transposon Tn916 was evaluated as a tool for generalized mutagenesis of the genome of This was achieved in by searching the genome sequence of Rd for the published Tn916 target site consensus sequence 5′ TT/ATTTT(N)AAAAAA/TA. This search identified 16 putative target sites. In subsequent experiments, integration of Tn916 did not occur at any of these sites. Using the nucleotide sequences of these observed integration sites, a new consensus sequence, 5′ TTTTT(N)AAAAA (4x7), was derived. This sequence reflects the curve-twist-curve DNA topology which is a feature common to all Tn916 integration sites. A search of the Rd genome using the new consensus sequence identified 167 potential target sites, representing approximately 1% of the total genome. Only 80 of these sites were located within ORFs. The presence of such a limited number of target sites places severe constraints on the use of Tn916 as a tool for generalized mutagenesis of the genome of


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