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. Author manuscript; available in PMC: 2016 Sep 1.
Published in final edited form as: Fungal Genet Biol. 2015 Jun 23;82:9–21. doi: 10.1016/j.fgb.2015.06.003

Figure 4. PCR and phenotypic validation of an insertion in the C. neoformans ACA1 gene (CNAG_05218) identified by AIM-Seq.

Figure 4

(a). Plot of the insertion site in the open reading frame of ACA1. Expanded view shows the region where insert flanking sequences align. Grey boxes indicate exons. (b). PCR confirmation of this site in strain WHM24. Primers flanking the putative insertion site were used to differentiate between wild type and mutated loci in AMT mutants. In lane 3, WHM24 shows a shift to a larger amplification product of 2.8 kb relative to the WT 500 bp product in lane 2. HyperLadder, 1kb, shown in lane 1 for reference (Bioline). (c). Phenotypic validation of WHM24. The sensitivity to pH 8 and growth defect on 1.5 M NaCl of the original AMT mutant, WHM24, are identical in an independent strain with an aca1Δ mutation.