ential gene expression analysis was carried out on P. pelagicus crabs from the following moult stages, post moult, intermoult, early pre moult, late pre moult and ecdysis, using custom prepared P. pelagicus cDNA microarray slides. A loop design, in which consecutive moult stages were compared via dual channel microarray hybridisations, was employed for the analysis. This Regorafenib format enabled the generation of a time series plot of differentially expressed genes across the moult cycle. Analysis with GeneSpring using K means clustering revealed seven main subsets of transcripts that displayed profiles and hence collectively termed Cluster B. Inhibitors,Modulators,Libraries Inhibitors,Modulators,Libraries Table 2 describes the composition of Cluster Inhibitors,Modulators,Libraries B, each subset of Cluster B is referred to individually.
Hemocyanin makes up 33% of the sequenced transcript population, crypto cyanin 23%, trypsin 15%, cathepsin 6%, chymotrypsin 5%, carcinin 3%, while fatty acid binding protein, dehy drogenase, ATP synthase, fumarase and arginine kinase each make up 2%. Additionally 6% Inhibitors,Modulators,Libraries of the sequenced transcripts remain unable to be annotated. Cluster C is a relatively small group of transcripts, in which expression is down regulated in the moult and post moult stages, increases substantially in differential expression profiles across the moult cycle of P. pelagicus. They were grouped into clusters labelled A G according to their expression patterns. Not all of the 556 clones selected for sequencing fell into these clusters and many of the transcripts associated with these clusters were not sequenced.
The transcripts assigned to Cluster A are relatively down regulated at the time of moulting, expression sub sequently increases at each consecutive stage, peaking in early pre moult then plateauing or declining slightly in late pre moult in preparation for ecdysis. In this GSK-3 subset, four clusters were deemed to have similar expression profiles and hence collectively termed Cluster A, this group is shown in Figure 3. Table 1 describes the composition of Cluster A, referring to each individual cluster respectively. The largest proportion of sequenced transcripts in Cluster A are of mitochon drial origin, 15% are metallothionein, 10% actin, 9% myosin, 8% opsin, 4% ferritin, while ribosomal RNA and elongation factor make up 2. 5% each, hemocyanin, chiti nase and a CT repeat sequence contribute to 1% of the transcripts sequenced from Cluster A.
Cluster B consists of transcripts which are down regu lated in the http://www.selleckchem.com/products/17-DMAG,Hydrochloride-Salt.html moult and post moult stages, expression then increases dramatically in intermoult, remains high in early pre moult and begins to decline in late pre moult. This group is depicted in Figure 3 where three clusters were deemed to have similar expression intermoult, then drops dramatically in early and late pre moult. Of the sequenced transcripts from Cluster C, three are cuticle proteins. The transcript profile that is represented in Cluster D displays expression that peaks in the post moult stage, decreases dramatically in the intermoult and