FIG. 4.

Different assembly rates for nucleoprotein complexes mediating full-site and half-site integration. (A) IN was assembled with the “G” 4.5-kbp donor at 14°C. At various time intervals (bottom), aliquots were taken and added to tubes containing target DNA at room temperature. Following strand transfer at 37°C, the products were subjected to agarose gel electrophoresis and quantified. (B) The same assembly experiment was repeated as described in panel A, except assembly was on ice. (C) The same assembly experiment was repeated as described in panel A at 14°C, except IN was at 10 nM and the donor was the wt 3.4-kbp DNA containing ∼30 bp of wt U5 and wt U3 sequences at its ends.