Eudiplozoon nipponicum (Monogenea): the molecules transcribed by hematophagous parasite of fish
| Autoři | |
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| Rok publikování | 2016 |
| Druh | Další prezentace na konferencích |
| Fakulta / Pracoviště MU | |
| Citace | |
| Popis | Ectoparasitic flatworms from the family Diplozoidae (Platyhelminthes, Monogenea) represent a serious bloodfeeding fish pathogens. Until now, the running research is focused mainly on morphological and phylogenetical characteristics of these worms and the information related to the biochemical and molecular nature of the physiological processes is rather sporadic. Therefore we started with a first complex transcriptomic followed by genomic analysis of monogenean representative - Eudiplozoon nipponicum Goto 1891 (Monogenea, Diplozoidae), which was performed by three sequencing strategies - 454/Roche, MiSeq Illumina and HiSeq Illumina. The mRNA of E. nipponicum adults, in the form cDNA was sequenced (454/Roche, MiSeq Illumina) and raw reads were processed in order to get high quality transcriptomics sequences for further annotation. Raw reads were filtered and low quality bases were removed as well as contaminations – reads related to host organisms (common carp). Sequencing errors and mismatches were corrected and finally the processed reads were assembled into the form individual transcripts. 454/Roche and Illumina sequences were pooled and several further bioinformatics approaches (especially searching for the closest homologous in non-redundant databases and prediction of proteins key properties) were used. We identified number of molecules of our interest (e.g. those related to hematophagy - proteolytic enzymes and their inhibitors, anticoagulant agents and immunomodulators). Some of these molecules are subjects of our further research. In order to reach the complete information of all E. nipponicum genes, the reads related to genomic DNA were generated by adoption of Illumina HiSeq platform. Quality of reads was checked, low quality reads were removed and total genome size and sequencing coverage were estimated. The estimation of genome size was performed also experimentally using DNA fluorescent double staining method, which represents simple and easy pointbased measurement of intensity of fluorescent signal given by amount of DNA. This method was originally designed for single-celled organisms and therefore we are optimizing the protocol for multicellular organism E. nipponicum. All obtained sequences on E. nipponicum have been compared with sporadic nucleotide sequential datasets from other members of the class Monogenea. |
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