Project description:Transcriptome profiling of wild type and cfo1 mutant with fluconazole treatment in Cryptococcus neoformans var. grubii H99 Purpose: The goals of this study are to compare cfo1 mutant transcriptome profiling (RNA-seq) to wild-type with or without fluconazole treatment in Cryptococcus neoformans var. grubii H99. Methods: mRNA profiles of wild-type and cfo1 mutant with or without fluconazole treatment were generated by RNA-Seq, using Illumina GAIIx. The sequence reads that passed quality filters were mapped to reference genome and the normalized RPKM values were calculated by CLC Genomics Workbench. Results: Compared to wild-type, a number of genes were differentially expressed in the cfo1 mutant, especially genes involved in iron homeostasis and transport, ergosterol biosynthesis, mitochondrial function and respiration. Conclusions: Our data suggested reduced expression of the genes in the respiratory chain is the main reason for altered antifungal sensitivity of the cfo1 mutant. The results of our study revealed that iron uptake plays a key role in fluconazole sensitivity of C. neoformans.
Project description:Transcriptome profiling of wild type and cfo1 mutant with fluconazole treatment in Cryptococcus neoformans var. grubii H99 Purpose: The goals of this study are to compare cfo1 mutant transcriptome profiling (RNA-seq) to wild-type with or without fluconazole treatment in Cryptococcus neoformans var. grubii H99. Methods: mRNA profiles of wild-type and cfo1 mutant with or without fluconazole treatment were generated by RNA-Seq, using Illumina GAIIx. The sequence reads that passed quality filters were mapped to reference genome and the normalized RPKM values were calculated by CLC Genomics Workbench. Results: Compared to wild-type, a number of genes were differentially expressed in the cfo1 mutant, especially genes involved in iron homeostasis and transport, ergosterol biosynthesis, mitochondrial function and respiration. Conclusions: Our data suggested reduced expression of the genes in the respiratory chain is the main reason for altered antifungal sensitivity of the cfo1 mutant. The results of our study revealed that iron uptake plays a key role in fluconazole sensitivity of C. neoformans. mRNA profiles of wild-type and cfo1 mutant with fluconazole treatment were generated by RNA-Seq, using Illumina GAIIx.
Project description:In Cryptococcus neoformans, H99 is the wild type. TJ1854 is the chromosome 4 disomy strain dervied from H99. In this study, approximately 1 million cells of TJ1854 were spread on YPD plate supplemented with 32ug/ml fluconazole. Randomly 27 adaptors (TJ3249-TJ3275) were chosen. These adaptors were sequenced.
Project description:In Cryptococcus neoformans, H99 is the wild type. TJ1843 is a strains with segmental disomy of chromosome 11, which is derived from H99. In this study, approximately 1 million cells of TJ1843 were spread on YPD plate supplemented with 32 ug/ml fluconazole. Randomly 27 adaptors (TJ3305 and TJ3331) were chosen. These adaptors were sequenced.
Project description:We measured protein translation (by ribosome profiling) and RNA levels (by polyA-enriched RNA-seq) in Cryptococcus neoformans strain H99 and Cryptococcus neoformans strain JEC21. This is the first transcriptome-wide map of translation in this species complex.
Project description:Invasive fungal infections (IFIs) are difficult to treat. Few effective antifungal drugs are available and many have problems with toxicity, efficacy and drug-resistance. To overcome these challenges, existing therapies may be enhanced using more than one agent acting in synergy. Previously, we have found amphotericin B (AMB) and the iron chelator, lactoferrin (LF), were synergistic against Cryptococcus neoformans and Saccharomyces cerevisiae. This study investigates the mechanism of AMB+LF synergy using RNA-seq in Cryptococcus neoformans H99.
Project description:Comparison of transcriptional profiles of WT Cryptococcus neoformans (H99) and strain CM126 (pRPL2b-GAT201) which overexpresses the transcription factor GAT201 using a ribosomal protein promoter Keywords: Genetic modification
Project description:Three analyses performed by the Duke Proteomics and Metabolomics Shared Resource in support of the project entitled "". Sample ID21519 was called Rra1-GFP, a 1D coomassie-stained gel band from a GFP-Trap pulldown of Rra1-GFP from the membrane protein fraction of Cryptococcus neoformans. Sample ID21570 was called no-GFP control, and consisted a GFP-Trap pulldown from the insoluble fraction of C. neoformans H99 lysate (non GFP expressing). ID21571 was called Rra1-GFP, and consisted of a GFP-Trap pulldown from the insoluble fraction of C. neoformans H99 lysate containing Rra1-GFP.
Project description:Cryptococcus neoformans lab strain H99 was spread on YPD plate supplemented with 4ug/ml benomyl. Randomly 60 adaptors were sequenced.
