Project description:Transcriptome analysis of Sodalis glossinidius derived from Trypanosoma brucei gambiense infection self cleared and infected Glossina palpalis gambiensis. At 3 time points (3, 10 and 20 days) after infectived blood meal, flies were analysed by PCR to isolate the infected and infection self cleared flies. Then, infected and infection self cleared flies midgut were dissected for RNA extraction.
Project description:Transcriptome analysis of Sodalis glossinidius derived from uninfected (controls) and Trypanosoma brucei gambiense infection self cleared Glossina palpalis gambiensis. 10 days after infectived blood meal, flies anal drop were analysed by PCR to isolate the infected self cleared flies. Then, uninfected (controls) and infection self cleared 10 days-flies midgut were dissected for RNA extraction.
Project description:Transcriptome analysis of Sodalis glossinidius derived from uninfected (controls) and Trypanosoma brucei gambiense infection self cleared Glossina palpalis gambiensis. 10 days after infectived blood meal, flies anal drop were analysed by PCR to isolate the infected self cleared flies. Then, uninfected (controls) and infection self cleared 10 days-flies midgut were dissected for RNA extraction. Total RNAs were extracted from 8 samples including: 4 control and 4 infection self-cleared flies.
Project description:Transcriptome analysis of Sodalis glossinidius derived from Trypanosoma brucei gambiense infection self cleared and infected Glossina palpalis gambiensis. At 3 time points (3, 10 and 20 days) after infectived blood meal, flies were analysed by PCR to isolate the infected and infection self cleared flies. Then, infected and infection self cleared flies midgut were dissected for RNA extraction. Total RNAs were extracted at 3 time points (3, 10 and 20 days) from 24 samples including, for each time, 4 infected and 4 infection self-cleared flies.
Project description:The majority of bacterial genomes have high coding efficiencies, but there are an few genomes of the intracellular bacteria that have low gene density. The genome of the endosymbiont Sodalis glossinidius contains almost 50% pseudogenes containing mutations that putatively silence them at the genomic level. We have applied multiple omic strategies: combining single molecule DNA-sequencing and annotation; stranded RNA-sequencing and proteome analysis to better understand the transcriptional and translational landscape of Sodalis pseudogenes, and potential mechanisms for their control. Between 53% and 74% of the Sodalis transcriptome remains active in cell-free culture. Mean sense transcription from Coding Domain Sequences (CDS) is four-times greater than that from pseudogenes. Core-genome analysis of six Illumina sequenced Sodalis isolates from different host Glossina species shows pseudogenes make up ~40% of the 2,729 genes in the core genome, suggesting are stable and/or Sodalis is a recent introduction across the Glossina genus as a facultative symbiont. These data further shed light on the importance of transcriptional and translational control in deciphering host-microbe interactions, and demonstrate that pseudogenes are more complex than a simple degrading DNA sequence. For this reason, we show that combining genomics, transcriptomics and proteomics represents an important resource for studying prokaryotic genomes with a view to elucidating evolutionary adaptation to novel environmental niches.
Project description:Tsetse flies (Glossina spp.) are major vectors of African trypanosomes, causing either Human or Animal African Trypanosomiasis (HAT or AAT). Several approaches are developed to control the disease among which the anti-vector Sterile Insect Technique. Another approach in the frame of anti-vector strategies could consist in controlling the fly’s vector competence which needs identifying factors (genes, proteins, biological pathways, …) involved in this process. The present work aims to verify whether protein candidates identified under experimental controlled conditions on insectary-reared tsetse flies have their counterpart in field-collected flies. Glossina palpalis palpalis flies naturally infected with Trypanosoma congolense were sampled in two HAT/AAT foci in Southern Cameroon. After dissection, the proteome from guts of parasite-infected flies were compared to that from uninfected flies in order to identify quantitative and/or qualitative changes associated to infection. A total of 3291 proteins were identified of which 1818 could be quantified. The comparative analysis allowed identifying 175 proteins with significant decreased abundance in infected as compared to uninfected flies, while 61 proteins displayed increased abundance. Among the former are RNA binding proteins, kinases, actin, ribosomal proteins, endocytosis proteins, oxido-reductases, as well as proteins that are unusually found such as tsetse salivary proteins (Tsal) or Yolk proteins. Among the proteins with increased abundance are fructose-1,6-biphosphatase, serine proteases, membrane trafficking proteins, death proteins (or apoptosis proteins), and SERPINs (inhibitor of serine proteases, enzymes considered as trypanosome virulence factors) that displayed highest increased abundance. Sodalis, Wiggleswothia and Wolbachia proteins are strongly under-represented, particularly when compared to data from similar experimentation conducted under controlled conditions on T. brucei gambiense infected (or uninfected) G. palpalis gambiensis insectary reared flies. Comparing the overall recorded data, 364 proteins identified in gut extracts from field flies were shown to have a homologue in insectary flies. Discrepancies between the two studies may arise from differences in the species of studied flies and trypanosomes as well as in differences in environmental conditions in which the two experiments were carried out. Finally, the present study together with former proteomic and transcriptomic studies on the secretome of trypanosomes, on the gut extracts from insectary reared and on field collected tsetse flies, provide a pool of data and information on which to draw in order to perform further investigations on, for example, mammal host immunization or on fly vector competence modification via para-transgenic approaches.