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. 1997 Jun 24;94(13):6851–6856. doi: 10.1073/pnas.94.13.6851

Figure 1.

Figure 1

Liberation of a LEU2 fragment and subsequent repair of the broken chromosome. (A) pJH825 is pBR322 plasmid containing URA3 inserted at the HindIII site, LEU2 surrounded by two 117-bp HO endonuclease recognition sites (cs), all inserted at the BamHI site, and a 1-kb internal fragment of the HIS4 gene inserted at the EcoRI site. When this plasmid is integrated at HIS4, it creates a His− duplication of part of the HIS4 gene (I). Expression of the HO endonuclease from a galactose-inducible promoter liberates the LEU2 fragment (II). The broken chromosome III is repaired by single-strand annealing to produce a His+ recombinant (III). The liberated LEU2 fragment may recombine with the leu2–3,112 allele at its normal chromosomal location approximately 22 kb away from HIS4 (IV). (B) Liberation and capture of a LEU2 fragment containing a 1.9-kb insertion of the ADE1 gene. Integration of this entire fragment into a wild-type LEU2 target produces a Leu+ Ade+ recombinant.