Project description:RNAsequening revolutionized the bacterial gene expression analysis. The objective of this study was to identify the genes involved in metabolism of Inulin in Ligilactobacillus agilis. We have obtained a list of genes upregulated in Ligilactobacillus agilis when it is grown in 1% Inulin
Project description:Proteins are ubiquitous macromolecules displaying a vast repertoire of chemical and enzymatic functions making them suitable candidates for chemosignals used in intraspecific communication. Proteins are present in skin gland secretions of vertebrates but their identity, and especially, their functions, remain largely unknown. Many species of lizards possess femoral glands, i.e. epidermal organs primarily involved in the production and secretion of chemosignals playing a pivotal role in mate choice and intrasexual communication. The lipophilic fraction of femoral glands has been well studied in lizards. In contrast, proteins have been the focus of only a handful of investigations. Here, we study the identity, inter-individual expression patterns and functionality of proteins present in femoral glands of sand lizards (Lacerta agilis) by applying mass-spectrometry proteomics. Our results showed that the total number of proteins varied substantially among individuals. None of the identified femoral gland proteins could be directly linked to chemical communication in lizards, although this result hinges on protein annotation in databases in which squamate semiochemicals are poorly represented. In contrast to our expectations, proteins consistently expressed across individuals were related to immune system, antioxidant activity and lipid metabolism as the main functions, adding support to the hypothesis that proteins in reptilian epidermal glands have other functions besides chemical communication. Interestingly, we found that major histocompatibility complex class I (MHC) expression is enriched in femoral gland secretions. Previously, MHC was hypothesized to have been coopted to serve a semiochemical function in sand lizards, specifically in partner recognition. We speculate with the possibility that MHC proteins could be linked to semiochemical function in sand lizards.
Project description:A laboratory colony of Phlebotomus perniciosus sand flies was maintained. Sand flies were infected with cultured Leishmania infantum promastigotes in stationary phase. Ten infected sand flies were dissected after 5 days and promastigotes within the gut pooled. The cells were immediately washed in PBS once and lysed in TRIzol reagent (Life Technologies). RNA isolation was completed according to the manufacturer's instructions, obtaining 63ng. RNA-seq libraries were generated using the spliced leader sequence for second strand synthesis (Cuypers et al., 2017; Haydock et al., 2015), thus allowing for specific amplification of sequences from L. infantum promastigotes, thus avoiding contamination with material from the sand fly gut. Single-end sequencing was performed in an Illumina HiSeq2500 instrument and data analysis was conducted using bowtie2, samtools, featureCounts and Geneious. The main findings are: i) substantial differences in differential gene expression between sand fly-derived (sfPro) and cultured (acPro) promastigotes; and ii) over-expression of genes involved in metacyclogenesis in sfPro vs. acPro, including gp63 genes, autophagy genes, etc.
Project description:In this study transcriptomic data of three life history stages of Orciraptor agilis was generated: 1) Gliding cells in absence of food ('gliding'), 2) Cells attached to the cell wall of its algal prey during perforation ('fattacking'), 3) Cells after acquisition of the algal plastid material ('digesting'). Furthermore, RNA-seq of the algal prey Mougeotia sp. was also performed. A de novo transcriptome assembly of the algal reads was performed in order to identify and substract algal reads of the Orciraptor samples by mapping the Orciraptor reads to the algal transcriptome. After this filtering step the remaining Orciraptor reads from all libraries were pooled for a de novo transcriptome assembly of Orciraptor agilis. This transcriptome was the basis for a comparative transcriptomic study in which transcript expression was compared between the three life history stages.