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. 2021 Apr 19;65(5):e01975-20. doi: 10.1128/AAC.01975-20

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Copyright © 2021 Leong et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 2 — Agrobacterium tumefaciens-mediated transformation (ATMT) of CYP51 mutation into wild-type CBS 7982 and mutant CBS 14141 M. furfur. (A and B) Phenotypes and genotypes of the M. furfur wild-type strain CBS 7982 (A) and the mutant strain CBS 14141 (B). (C and D) Vector maps for CBS 7982 (C) and CBS 14141 (D) ATMT. (E) Screening and selection of CBS 7982 transformant colonies with the correct CYP51 insert (∼700 bp) and 1.5-kb upstream/downstream flanking arms. (F and G) Sanger sequencing validation of the correct single nucleotide mutation and ITS sequences in CBS 7982 (F) and CBS 14141 (G). (H) Heat map of the MICs for CBS 7982 Y67F and CBS 14141 F67Y mutants. MICs were normalized from 0 to 1, with 1 being the highest MIC (red).