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. 2004 Nov;24(21):9305–9316. doi: 10.1128/MCB.24.21.9305-9316.2004

FIG. 3.

FIG. 3.

Expression of RAD51-K133R increases gene conversion of markers distant from the DSB during HDR. (a) Structure of the H-DR-8mu reporter and summary of gene conversion tracts in neo+ recombinants. The pneo-8mu fragment contains the wild-type NcoI site at the site of the DSB, along with 1-bp silent mutations, which create the following restriction site polymorphisms: A, ApaI; L, ApaL1; P, PstI; B, BamHI; X, XbaI; Nr, NruI; and Pm, PmlI. HDR of the I-SceI-generated DSB in S2neo, using the pneo-8mu gene as the template for repair, results in restoration of a wild-type neo+ gene, so that all recombinants have an NcoI site incorporated. HDR may be associated with incorporation of the other restriction sites. The filled bars represent the incorporation of all restriction site markers up to and including the indicated site for various recombinants derived from the parental cell line or after expression of RAD51-WT or RAD51-K133R. (b) Summary of gene conversion frequencies of each restriction site marker for the recombinants shown in panel a. The conversion frequency for each restriction site is graphed as a function of distance from the DSB.