Fig. 4.

A novel mutation, T297M, detected by direct sequencing of the CYP51 PCR product of T. cruzi Y strain H10-R clones. The T297M mutation is associated with broad-spectrum resistance to CYP51 inhibitors. Left panel: the codon for amino acid 297 (boxed) is mutated from ACG in the wild-type to AYG in the resistant clones, where Y = C + T. Right panel: the previously described mutation in the codon for amino acid 355, which would change CCA to TCA, (boxed) was not present. The TcCYP51 genes were sequenced of sensitive (H10-LT) and the resistant (H10-R 500 nM ravuconazole and H10-R-NP) clones, along with other clones that were isolated, either earlier (H10- 4 nM and H10-R 8 nM) or later (H10-R 1 μM) during the process of in vitro selection of resistance.