Cryptococcus neoformans gene expression in minimal media of wild type (WT) and ALL2 mutant (all2Δ) cells

To examine the comparative transcription profile of WT and all2Δ mutant cells in in vitro conditions to determine genes responsible for hypervirulent phenotype of all2Δ cells. Using transcriptional profiling, we determined that many genes involved in transport were differentially transcribed between the wild type and the mutant strain. With the help of further experiments we could rule out that ALL2 has a unique function in maintaining the intracellular pH under acidic conditions. For arrays, total RNA was isolated from WT and all2Δ mutant using Qiagen RNeasy Kit. For RNA extraction, WT and all2Δ mutant were grown in minimal media. Cells were grown for overnight, diluted 1:100 and then again grown overnight or up to an OD of 0.6-0.7 with agitation (150rpm) at 37°C.