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Fig. 8. Gap repair. (A) Recombineering-mediated gap repair. Two
homology arms, located at the 5' (Left, L) and 3' (Right, R) end
of a genomic region of interest present in a BAC, are cloned into the desired
plasmid. Restriction enzyme-mediated linearization between both homology arms
and subsequent transformation in bacteria competent for recombineering
functions allow the selective retrieval of the desired fragment from the BAC
into the plasmid through gap repair. The resulting plasmid can be used for
P transposase- (5'P and
3'P element termini) or 
C31 integrase-mediated
transgenesis (attB site). (B) In vivo gap repair. Two homology
arms, located at the 5' and 3' ends of a genomic region of
interest, are cloned within a P element. After P
transposase-mediated germ line transmission, the transgene is linearized in
vivo between both homology arms using the meganuclease I-SceI, potentially
resulting in the selective capture of the desired fragment from a wild-type
chromosome through homologous recombination-mediated gap repair.
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