Gene Expression Arrays comparing C. dubliniensis CEM002 during planktonic and biofilm growth in different media

The goal was to identify genes that change their expression in biofilm vs. planktonic growth. Cultures for the extraction of total RNA under biofilm growth conditions were performed on biofilms grown on the bottom of 6-well polystyrene plates for 48 hours at 37°C and 200 rpm as previously described (Nobile et al. 2012). The media used was Spider 1% glucose and Spider. Planktonic cultures for total RNA were grown in the same media by inoculating with cells from an overnight 30°C YPD culture to an OD600 of 0.05. Cultures were then grown in flasks at 37°C with shaking at 225 rpm until they reached an OD600 of 1.0. Expression was measured relative to pooled references, two replicates of each experiment were performed.

本研究旨在鉴定在生物膜（biofilm）与浮游（planktonic）生长状态下表达水平发生改变的基因。生物膜生长条件下总RNA提取所用的培养物，参照此前报道（Nobile等，2012），由37℃、200 rpm条件下在6孔聚苯乙烯培养板底部生长48小时的生物膜制备得到，所用培养基为Spider 1%葡萄糖培养基与Spider培养基。用于总RNA提取的浮游培养物，采用经30℃过夜培养的YPD培养基中的细胞接种至OD600为0.05的初始浓度，采用与生物膜培养相同的培养基进行培养；随后将培养物置于摇瓶中，于37℃、225 rpm振荡培养至OD600达到1.0。基因表达量以混合参照样本作为相对定量基准，每个实验均设置两次重复。