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. 2009 Oct 6;284(48):33683–33691. doi: 10.1074/jbc.M109.044024

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© 2009 by The American Society for Biochemistry and Molecular Biology, Inc.

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FIGURE 3. — Disruption of the native Der1_Ap locus. a, Der1_Ap locus was disrupted by homologous recombination using a targeting construct carrying a CAT marker flanked by 2 kb of gene-specific upstream and downstream sequences. Recombinants were isolated using chloramphenicol to select for CAT expression, and cell sorting to select against YFP expression. Wild type (RH-WT), parental (carrying the native (Der1_Ap) as well as an inducible minigene version (iDer1_Ap), and ΔDer1_Ap (carrying only the inducible version) were tested for the presence of the native locus by b, PCR (position of primers is indicated in a), or c, Southern blot using the radiolabeled coding sequence of Der1_Ap as probe and Nde1 restriction to distinguish the native and inducible locus.