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. 2010 Feb 1;1:8. doi: 10.1186/1759-8753-1-8

Figure 2.

Figure 2

Immunity of the linear Mu genome in vitro. (a) Transposition reactions were set up by incubating MuA, MuB and HU proteins with the mini-Mu plasmid pSP104 as donor and either linear Mu genome or pUC19 as target, as described in Methods. Lane 1, cleaved Type I complex assembled on pSP104; lanes 2 to 4, Type II or strand transfer reactions with: pSP104 as donor and either pUC19 (lane 2) or Mu (lane 3) as target or Mu as donor and pUC19 as target (lane 4); lanes 5 to 7, control substrates without added proteins. (b) Map of pSP104 showing restriction enzyme sites used for linearization and their position with respect to the attL and attR Mu ends. (c) Transposition reactions with linear mini-Mu as target. Reaction conditions were as in (a), except that Type I complexes were first assembled on pSP104 and added to indicated targets in a second step. Lane1, Type I reaction; Lane 2 to 7, Type II reactions. Lanes 8 to 13, DNA controls without added proteins. Restriction enzyme shown in the B panel are abbreviated to H, X and B. L = linear. O = open circular; S = supercoiled; Type I = cleaved complex; Type II = strand transfer complex; Type II (intra) = intramolecular strand transfer complexes.