Project description:Transcriptional profiling of Leishmania major parasites overexpressing LmSir2rp3 protein compared with wild-type cell line transfected with empty vector in normal growth conditions. Goal of this experiment was to evaluate the possible effect of LmSir2rp3 in the control of gene expression in this protozoan parasite that has a polycistronic transcription. Two-condition experiment, LmSir2rp3 overexpressor vs. Wild-type cell line bearing empty vector. Biological replicates, 3 LmSir2rp3 overexpressor replicates and 3 wild-type cell line bearing empty-vector
Project description:Transcriptional profiling of Leishmania major parasites overexpressing LmSir2rp3 protein compared with wild-type cell line transfected with empty vector in normal growth conditions. Goal of this experiment was to evaluate the possible effect of LmSir2rp3 in the control of gene expression in this protozoan parasite that has a polycistronic transcription.
Project description:The genomic DNAs of strains JPCM5 and 263 of L. infantum, strains LV39 and Friedlin of L. major and strains Parrot-TarII and S125 of L. tarentolae were used in comparative genomic hybridizations to reveal the intra-species and inter-species gene content, and to validate L. tarentolae Parrot-TarII genome sequencing results. Leishmania (Sauroleishmania) tarentolae was first isolated in the lizard Tarentola mauritanica. This species is not known to be pathogenic to humans but is often used as a model organism for molecular analyses or protein overproduction. The Leishmania tarentolae Parrot-TarII strain genome sequence was resolved by high-throughput sequencing technologies. The L. tarentolae genome was first assembled de novo and then aligned against the reference L. major Friedlin genome to facilitate contig positioning and annotation, providing a 23-fold coverage of the genome. This is the first non-pathogenic to humans kinetoplastid protozoan genome to be described, and it provides an opportunity for comparison with the completed genomes of the pathogenic Leishmania species. A high synteny was observed in de novo assembled contigs between all sequenced Leishmania species. A number of limited chromosomal regions diverged between L. tarentolae and L. infantum, while remaining syntenic with L. major. Globally, over 90% of the L. tarentolae gene content was shared with the other Leishmania species. There were 250 L. major genes absent from L. tarentolae, and interestingly these missing genes were primarily expressed in the intracellular amastigote stage of the pathogenic parasites. This implies that L. tarentolae may have impaired ability to survive as an intracellular parasite. In contrast to other Leishmania genomes, two gene families were expanded in L. tarentolae, namely the leishmanolysin (GP63) and a gene related to the promastigote surface antigen (PSA31C). Overall, L. tarentolae appears to have a gene content more adapted to the insect stage rather than the mammalian one. This may partly explain its inability to replicate within mammalian macrophages and its suspected preferred life style as promastigote in the lizards.