FIGURE 5.

In vitro transposition efficiency of Mos1 and Mboumar-9 transposons flanked with different IRR and IRL combinations. A, schematic of the in vitro transposition assay. The donor plasmid is incubated with a target plasmid, containing an ampicillin resistance (ampR) gene and a colE1 origin of replication, and purified transposase. The donor plasmid has a conditional origin of replication (oriR6K) and is unable to replicate in the recipient strain E. coli DH10B. The products of transposition are scored by counting the number of colonies carrying the kanR marker. The transposition efficiency was calculated as the number of kanR colonies per 1 μg of the donor plasmid divided by the transformation efficiency (CFU/μg). B, relative in vitro transposition efficiencies of Mos1 and Mboumar-9 donor plasmids containing different combinations of left and right IRs (as in Fig. 3B). These experiments were performed using MgCl₂. For ease of comparison, the in vitro transposition efficiency was normalized to 1.4 × 10⁻⁴, the transposition efficiency of the Mboumar-9 pEPMboRR donor plasmid. Each experiment was conducted 4 times, and the error bars indicate the S.D. between measurements. LL, two left IRs; LR, one left and one right IR; RR, two right IRs.