Project description:In Europe, ticks are the most important vectors of diseases threatening humans, livestock, wildlife and companion animals. Nevertheless, genomic sequence information and functional annotation of proteins of the most important European tick, Ixodes ricinus, is limited. Here we present the first analysis of the I. ricinus genome and of the transcriptome of the unfed I. ricinus midgut. We combined and integrated data from genome, transcriptome and proteome. The de novo assembly of 1 billion paired-end sequences identified 6,415 putative genes providing an unprecedented insight into the I. ricinus genome. Mapping of our midgut mRNA reads to the assembled contigs let us estimate to cover around two third of the unique genomic sequences. In addition, more than 10,000 transcripts from naïve midgut were annotated functionally and/or locally. By combining the alignment-based with a motif-search based annotation approach, we could double the number of annotations throughout all groups without shifting the dataset. Moreover, 1,175 proteins expressed in the naïve midgut were identified by mass spectrometry confirming the high completeness of our transcriptome database, and 608 were significantly annotated for function and/or localization. This multiple-omics study vastly extends the publicly available DNA, RNA and protein databases for I. ricinus and ticks in general.
Project description:Here, we challenged rabbits with repeated feeding of Ixodes ricinus adults and observed the formation of specific antibodies against several tick salivary proteins. To identify the salivary antigens, isolated immunoglobulins from repeatedly infested rabbits were utilised for a pull-down from the saliva of pilocarpine-treated ticks. Eluted antigens were separated on 1D SDS-PAGE and analysed by peptide mass fingerprinting. To increase the authenticity of immunogens identified, we also performed, for the first time, de novo assembly of the sialome from I. ricinus females fed for six days, a timepoint used for pilocarpine-salivation.