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. 2005 Feb;73(2):1239–1242. doi: 10.1128/IAI.73.2.1239-1242.2005

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Copyright © 2005, American Society for Microbiology

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FIG. 4. — CaNAT1 can mark a recyclable cassette for deletion of both alleles of a C. albicans gene. (A) Genomic DNA of the following strains was digested with NsiI: lane 1, wild-type SC5314; lane 2, JKC377, and lane 3, JKC378 (CaMOS10/Camos10::CaNAT1-FLP cassette); lane 4, JKC380, and lane 5, JKC382 (CaMOS10/Camos10::FRT); lane 6, JKC390, and lane 7, JKC392 (Camos10::FRT/Camos10::CaNAT1-FLP cassette); and lane 8, JKC393, and lane 9, JKC395 (Camos10::FRT/Camos10::FRT). The CaNAT1-FLP cassette is FRT-p-SAP2-FLP-t-CaACT1-p-AgTEF1-CaNAT1-t-AgTEF1-FRT. Left arrows show locations of molecular weight markers. Right arrows show genomic fragments. (B) Cartoon of the CaMOS10 locus, the deletion construct, the construct to reintegrate an intact CaMOS10 allele, and the probe used in Southern blotting. ORF, open reading frame. (C) Genomic DNA of the following strains was digested with NsiI: lane 1, wild-type SC5314; lane 2, JKC393, and lane 3, JKC395 (Camos10::FRT/Camos10::FRT); and lanes 4 and 5, transformants of JKC393 (lane 4, JKC504, and lane 5, JKC505), and lanes 6 and 7, transformants of JKC395 (lane 6, JKC506, and lane 7, JKC507) (Camos10::FRT/Camos10::FRT/CaMOS10 CaNAT1).