Comparison of in vitro DNA cleavage and DNA transposition of the donor Mboumar-9 transposons pEPMboLL (with two native IRLs) and pEPMboLL-G3′A (with two mutated IRLs).
A, agarose gel of the products of in vitro DNA cleavage. Experiments were performed twice. Lane 1, 1-kb DNA ladder of markers (M); lane 2, pEPMosLL linearized with XbaI; lane 3, pEPMboLL digested with SacI to excise the transposon; lane 4, pEPMboLL plasmid; lane 5, pEPMboLL plasmid incubated with Mboumar-9 transposase; lane 6, pEPMboLL-G3′A plasmid; lane 7, pEPMboLL-G3′A plasmid incubated with Mboumar-9 transposase. LL, two left IRs. B, quantification of in vitro cleavage and transposition of these transposons. Three repeats of the transposition reactions were performed. The percentage of backbone DNA released from the LL and LL-G3′A donor plasmids, as well as the relative efficiency of Mboumar-9 in vitro transposition, is normalized to that of LL. The error bars indicate S.D. between multiple measurements.