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. 2001 Jul;13(7):1609–1624. doi: 10.1105/TPC.010168

Figure 2.

Figure 2.

Analysis of Engineered hdc1 Mutants

(A) Restriction map of the genomic region of wild-type HDC1 and predicted map of the gene replacement mutant. The open box in the wild-type map indicates the fragment of HDC1 DNA replaced by the hph gene encoding hygromycin phosphotransferase in the mutant. The arrow indicates the location of the HDC1 coding region. B, BamHI; H, HindIII; P, PstI; C, ClaI; X, XhoI.

(B) DNA gel blot probed with hph. Lane 1, wild-type strain 367-2. Lanes 2 to 11, 10 independent transformants: lane 2, T702.1; lane 3, T702.2; lane 4, T702.3; lane 7, T702.4; lane 8, T702.5.

(C) The same blot probed with the deleted segment of HDC1. On the basis of the hybridization pattern, all of the transformants except those shown in lanes 5 and 6 have undergone gene replacement by simple double crossover homologous integration. Sizes of DNA markers in kilobases are shown at right.