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. 2006 Feb;72(2):1645–1652. doi: 10.1128/AEM.72.2.1645-1652.2006

FIG. 3.

FIG. 3.

Complementation of pigmentation in the G. zeae ΔGIP2 strain. (A) Complementation strategy with GIP2 from G. zeae. ΔGIP2, genomic DNA from the GIP2-deleted strain of Z03643 (Tzg2-1); Rg2-3, genomic DNA of the transgenic Tzg2-1 strain carrying GIP2; K, KpnI; hygB, hygromycin resistance gene; gen and amp, genes conferring resistance to geneticin and ampicillin, respectively. GzORF1 and GIP3 are indicated by open arrows. (B) Pigmentation of the transgenic strains examined by DNA gel blot analysis. Left, Z03643; middle, Tzg2-1; right, Rg2-3. (C) DNA gel blots of transformants derived from the ΔGIP2 strain by using an intact copy of the GIP2 ORF. Genomic DNAs of these transformants were digested with KpnI and hybridized with the probes indicated in panel A. The probe for hybridization with GIP2 was amplified from genomic DNA of Z03643 with primers G2-for and G2-rev (Table 1). Lane 1, wild-type strain Z03643; lane 2, ΔGIP2 recipient strain Tzg2-1; lane 3, transformants (Rg2-3) carrying GIP2. The sizes of λDNA standards (in kilobase) are indicated on the left of the blot. (D) RNA gel blot of strain Z03643 and the transgenic ΔGIP2 strain probed with GIP2. Z03643 and ΔGIP2, total RNAs from wild-type strain and the ΔGIP2 recipient strain Tzg2-1, respectively; Rg2-3, total RNAs from transgenic Tzg2-1 strains carrying GIP2. The probe and the incubation time are indicated on the left and above, respectively. The ethidium bromide-stained rRNAs are indicated as a loading control.