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Figure 3 | BMC Microbiology

Figure 3

From: Molecular characterization of KU70 and KU80 homologues and exploitation of a KU70-deficient mutant for improving gene deletion frequency in Rhodosporidium toruloides

Figure 3

KU70 deletion strategy and Southern blot results. (A) Schematic illustration of KU70 deletion strategy. LB and RB are the left border and right border sequences of T-DNA derived from pPZP200, respectively; P GPD1 : R. toruloides GPD1 promoter; hpt-3: codon-optimized hygromycin phosphotransferase gene; T nos : transcriptional terminator of A. tumefaciens nopaline synthase gene; LoxP: recognition sequences of Cre recombinase; Rg70Lf and Rg70Rr: primers to amplify KU70 gene deletion region; Rg70f3 and Rg70r2: primers for fungi colony PCR; Rt100 and Rt101: primers to amplify probe used for Southern blot analysis. Unique restriction enzyme digest sites used are shown. (B) Southern blot results of putative ∆ku70 transformants. Genomic DNA was digested with Pvu I and a band shift from 2.2 kb (WT) to 2.7 kb indicates successful deletion of KU70.

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BMC Microbiology

ISSN: 1471-2180

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