Figure 7.
Effect of reassociation of INZn on in vitro integration activity. Assays were performed with 100 ng receptor DNA and 10 ng pre-processed (A) or unprocessed donor (C) with 1 pmol of INzn 12.5μm or 0.125 µM diluted in standard dilution buffer (7.5 mM MnCl2, HEPES 20 mM, pH 7.6) without preincubation (respectively lanes 1 and 2) or after preincubation with 1 pmol of specific 21 bp viral ODN (lanes 3) or random 21 bp ODN (lane 4) under standard conditions (7.5 mM MnCl2, HEPES 20 mM, pH 7.6). The reaction products were then loaded on 1% agarose gel. The position and the structure of the different products obtained after half-site (HSI), full-site (FSI) and donor/donor integration (d/d) are reported in the figure. Effect of reassociation on full site integration. Reaction products obtained with the pre-processed (B) or the unprocessed (D) donor were cloned in MC1060/P3 E. coli strain as described in Materials and methods section. The number of resistant selected colonies obtained after integration reaction under the different conditions is reported in the figure. Results are the mean ± SD (error bars) of three independent experiments.