Detection of ERG11 gene mutation in coding and non-coding region of clinical Candida glabrata isolates from Pakistan, a short communication
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Azoles inhibit cytochrome P450 dependent enzyme lanosterol-14α-demethylase (CYP51) that is encoded by ERG11gene. Mutations in the ERG11gene confer azole resistance in Candida species through different mechanisms, like ERG11 overexpression, CDR1,2 overexpression, tolerance to methylation, biofilm formation and import of host cholesterol. We sequenced ERG11 gene to determine mutations in coding and non-coding region of ERG11 in clinical isolates of Candida glabrata from Pakistan. Eight C. glabrata strains with fluconazole minimum inhibitory concentrations ranging from 2-256 mg/L were included in this study. The ERG11 gene was amplified by PCR and sequenced using Sanger methodology and analyzed using bioinformatic tools. We identified non-synonymous mutation at position I166S in non-coding region of ERG11gene in C. glabrata. Two novel synonymous mutations at position I100 and D278, and previously reported synonymous mutations D256, L341, P519 and R527 were also observed. This is the first study evaluating ERG11 mutations in C. glabrata species from Pakistan. In this study, we reported I166S mutation in non-coding region of ERG11 gene. Such ERG11 gene mutations which are unique to geographical regions should be investigated through resistance and genotypic surveillance.
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Thank you for submitting your manuscript to Access Microbiology. The study is interesting for the understanding of regional fuconazol resistance in C. glabrata. However, a major revise is needed before progressing to peer-review. The methods need a more comprehensive description and a division in different subsections for each experiment. The results could be further described (for example, the phylogenetic analysis). The DNA sequences obtained throughout the work need to be made publicly available in a related repository and the respective reference should be cited in the Data availability section. Figure 2 is included in supplementary material needs to be cited in the main text. Figures included in the supplementary material need to be named accordingly (Supplementary Figure S...). Can a mutation occurring in in a non-coding region …
Thank you for submitting your manuscript to Access Microbiology. The study is interesting for the understanding of regional fuconazol resistance in C. glabrata. However, a major revise is needed before progressing to peer-review. The methods need a more comprehensive description and a division in different subsections for each experiment. The results could be further described (for example, the phylogenetic analysis). The DNA sequences obtained throughout the work need to be made publicly available in a related repository and the respective reference should be cited in the Data availability section. Figure 2 is included in supplementary material needs to be cited in the main text. Figures included in the supplementary material need to be named accordingly (Supplementary Figure S...). Can a mutation occurring in in a non-coding region of a gene be synonymous or non-synonymous if that region would not be translated to a protein? The use of English and the grammar need an exhaustive revision (please contact the Editorial Office if you would like to explore English proofing options). Please provide a revised version of the manuscript (including a tracked-changes document) addressing these comments, along with a point-by-point response to them within 2 months.
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