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. The primary role of the perisotracum is believed to be shell secretion and T0901317  mainte nance, T0901317  but there are other roles such as protection from the external environment, infestation and predatory GANT61 bor ers, The mantle tissue for this analysis was taken as a cross section of all mantle folds and included periostracum material. Human musculoskeletal system So far molecular work in L. elliptica has been limited to candidate genes concerned with understanding thermal tolerance via antioxidants and heat shock proteins, Truebano et al, submitted]. The aim of this study is to develop molecular resources for this species. We focus on the mantle, as it is the main shell secreting organ and we are particularly interested in the processes of calcification and comparative ossification, 454 pyrosequencing technologies enable the rapid generation of transcriptomes for non model species, This approach is exploited in the present study, the results of which will be the basis of future studies of calcium regu lation in L.
Lomeguatrib elliptica in relation to environmental change. Here we describe the transcriptome of the mantle tissue of L. elliptica, focussing on the datamining of genes involved in calcium regulation and shell deposition. This represents the first publicly available 454 data for an Ant arctic marine invertebrate and provides an important comparative resource for such studies in more euryther mal temperate mollusc species. Results and Discussion The non normalised laternula libraries were subjected to a full 454 run that yielded 1,034,155 reads totalling 381,838,384 bases with an average read length per tran cript of 369 bases.
After cleaning the data and removing small reads, 778,629 reads with an average size of 304 bp were entered into Newbler for assembly. These assem bled T0901317  into the 18,290 contigs which were used for further analysis. Because the aim of this project was to identify and characterise specific genes for future analyses, in particular the GPCRs and several gene fami lies, such as collagen and bone morphogenic proteins there was a require ment for longer sequences of good quality which would enable us to distinguish between gene family members. Hence the descriptive analysis presented here utilised only the contigs produced by the assembly.
Whilst the singletons potentially contain useful lowly expressed sequences, they also contain a substantial proportion of artefacts derived from cDNA synthesis, sequencing and contamination, PCR and re sequencing of singletons is essential Lomeguatrib in order to verify the gene products, The contigs ranged in size from 142 bp to 5591 bp, with an average size of 535 bp. 42 contigs were greater than 3 kb and 69 contigs comprised more than 300 reads, with the largest contig of 5591 bp comprising the most reads with 1000 sequences, Self BLAST of this dataset pro duced only 281 matches with a value of e 100, indicating a low level of redundancy in the assembly of the reads. The contigs contained 46,235 microsatellites, of which 1,608 comprised over 7 repeat units, There were 13,084 SNPs INDELS present in 2,475 contigs designated as high confidence by the Newbler program, although a further circa 25,000 SNPs were identified at lower confi dence level as defined by Newbler, These figures for microsatellite and SNP INDEL detection are at a similar relative level to those identified in the transcriptome of another non model species, the flesh fly, Given that L.
elliptica is a wild caught species and the S. crassipalpis material came from a long standing inbred laboratory stock, higher levels of these T0901317  genetic variants would be expected in the clam. However, analysis in this species was restricted to contigs only, potentially reducing the dataset. Sequence similarity Lomeguatrib searching of the GenBank non redundant database with BLAST produced matches against only 3,098 of the contigs using a 1e 10 cut off value. This poor level of sequence similarity matching has also been noted in previous work on this spe cies and is due to a lack of sequences from