BioProject

To investigate the systemic molecular changes occurring as a result of Dhr96 knockdown or over-expression, a comparison between knockdown or overexpression lines and their genetic controls were performed. More...

To investigate the systemic molecular changes occurring as a result of Dhr96 knockdown or over-expression, a comparison between knockdown or overexpression lines and their genetic controls were performed. 0-3 day old adult males or females were reared on 3 separate batches of diet (this was the standard diet we used for culturing Drosophila melanogaster and was made up of 10L water, 100g agar (USP #7060 Bio-serve), 350g Brewer’s dried yeast (Sunshine Health), 300g black treacle (Lyles), 150g sucrose (Tate & Lyle), 300g Difco dextrose (Becton Dickinson), 150g cornmeal (#1151, Bioserve), 100g wheatgerm (#1659, Bioserve), 200g soya bean flour (#S9633 Sigma Aldrich), 10g methyl-4-hydroxybenzoate (#H3647 Sigma Aldrich) in 10ml ethanol, 50ml proprionic acid (#P5561 Sigma Aldrich)). Each of these 3 batches was considered to represent independent biological replication. The RNA samples were hybridized to the Affymetrix Drosophila GeneChip 2. Previous Q-PCR analysis had indicated that there was differential Dhr96 gene expression in the knockdown or overexpression strains compared to their controls. (This was also inferred from phenotypes resulting from other distinct crosses.) However the first analysis carried out on these samples was to verify the levels of differential Dhr96 gene expression. From this analysis we found no evidence that Dhr96 is down-regulated in the knockdown line compared to its control for either Males or Females, as the confidence intervals for the fold change include the value of one and the p-values are large. Therefore the knockdown line and its control samples were excluded from further analysis. The overexpression line did however show significant up-regulation of Dhr96 expression relative to its control in both Males and Females, and was further analyzed. Overall design: Flies of the appropriate genotype (Dhr96 knockdown or over-expression and their genetic controls) were reared on 3 different batches of diet (same recipe, prepared on different days). 0-3 day old adults were collected (males and females stored seperately) and each sample was hybridised independently to a chip. Less...