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

FIG. 5.

FIG. 5.

Overexpression of GIP2 in G. zeae ΔGIP2 strains. (A) Overexpression strategy. ΔGIP2, genomic DNA of Tzg2-1, the ΔGIP2 strain derived from Z03643; Odzg2-1, genomic DNA of Odzg2-1, the transgenic Tzg2-1 strain carrying the GIP2 gene under control of the G. zeae β-tubulin gene promoter; pSK660, a vector used for cotransformation; K, KpnI; Pb-tub, promoter region of the β-tubulin gene from Z03643; hygB, hygromycin resistance gene; gen and amp, genes conferring resistance to geneticin and ampicillin, respectively. The probe used for blot hybridization, which is amplified from genomic DNA of Z03643 with primers G2-for and G2-rev (Table 1), is indicated by a thick bar. GzORF1 and GIP3 are indicated by open arrows. (B) DNA gel blot of transformants derived from ΔGIP2 strains, hybridized with GIP2. Lane 1, Z03643; lanes 2, Tzg2-1, the ΔGIP2 recipient strains derived from Z03643; lanes 3, Odzg2-1, a transformant derived from Tzg2-1. The sizes of λDNA standards (in kilobases) are indicated on the left of the blot. (C) Pigmentation of transgenic strains. The upper and lower plates in each panel indicate the strains derived from Z03643 and SCKO4, respectively. WT, wild-type strains; ΔGIP2, the transgenic strains with GIP2 deleted (Tzg2-1 and Tsg2-2 derived from Z03643 and SCKO4, respectively); Odg2, the GIP2-overexpressing transformants from the ΔGIP2 strains (Odzg2-1 and Odsg2-2 derived from Tzg2-1 and Tsg2-2, respectively).