Project description:Amplification of small subunit rRNA sequences from Brazilian clinical isolates of Trypanosomatidae spp.
| PRJEB25557 | ENA
Project description:Amplification of glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) partial sequences from promastigote clones of Brazilian clinical isolates of Trypanosomatidae sp.
| PRJEB33769 | ENA
Project description:Amplification of tubulin partial sequences from Brazilian clinical isolates of Trypanosomatidae sp.
| PRJEB25558 | ENA
Project description:Amplification of ribosomal internal transcribed spacer 1 (ITS1) sequences from Brazilian clinical isolates of Trypanosomatidae sp.
Project description:Leishmania infantum (Kinetoplastida:Trypanosomatidae) is the etiological agent of zoonotic visceral leishmaniasis in the Mediterranean basin. The motile promastigote stage infects the hematophagous sand fly vector host and amastigotes survives and multiplies within phagocytes of the mammalian host. Promastigotes are routinely cultured in liquid undefined media and are considered to mimic the environment within the sand fly gut. We have put this to the test by high-throughput gene expression profiling by shotgun DNA microarrays generated in our laboratory. This has been possible thanks to RNA amplification.
Project description:The DEAD/H RNA helicase LINF_220021200 (DEVH1) gene from Leishmania infantum (Kinetoplastida:Trypanosomatidae) was cloned in the pTEX expression plasmid vector for trypanosomatids. Leishmania infantunm promastigotes were transfected and a knock-in L. infantum promastigote cell line was selected with geneticin (G418). A pTEX control promastigote line was also generated. Then, three independent biological replicate cultures of each pTEX-DEVH1 and pTEX promastigote lines were performed in the presence of the selective agent. The parasites were harvested on day 7 (stationary phase). Total mRNA samples were obtained. Cyanine dye-labelled samples were obtained from the knock-in and the control line (Cy5 and Cy3, respectively) and they were hybridized with custom whole-genome L. infantum DNA microarrays. This platform is included in GEO (GPL6781) and has also been repeatedly used in different experiments from 2009. Hybridization analysis allowed for finding differentially expressed genes due to the effect of induced over-expression of the DEVH1-encoding gene in the knock-in promastigote line compared to the control line. Genes involved in parasite infectivity and survival such as the HASP/SHERP gene cluster and an amastin gene or redox homeostasis genes are significantly down-regulated in the pTEX-DEVH1 knock-in promastigote line, whereas genes related to growth are up-regulated. This is in agreement with previous experimental data supporting that L. infantum DEVH1 knock-in promastigotes are able to recover the growth rate when stress conditions are removed.
Project description:Amplification of small subunit rRNA sequences of trypanosomatids from human samples of a patient diagnosed with visceral leishmaniasis.
Project description:Mgf files of the analysis of 66 extracts of 22 strains of Streptomyces spp. actinobacteria isolated in association to Anthurium spp. in brazilian oceanic islands, plus culture media blanks.
Project description:Methylation differences between SIC 5 cocoa seedlings germinated in a common environment before growth for 74 days under semic controlled environmental conditions set to mimic the Malaysian or Brazilian temperature environment. Genomic DNA sonicated and the methylated fraction enriched using a 5-methylcytosine antibody before whole genome amplification and hybridisation to the array. Two-condition experiment, Methyl enriched gDNA Malaysian vs. Brazilian leaf samples. 7 Biological replicates independently grown and harvested. Environmental (temperature) effect
Project description:An Easy Operating Pathogen Microarray (EOPM) was designed to detect almost all known pathogens and related species based on their genomic sequences. For effective identification of pathogens from EOPM data, a statistical enrichment algorithm has been proposed and further implemented in a user-friendly interface. A microarray was designed with probes for vertebrate-infecting virus sequences in EMBL, 18S rRNA fungi and parasite sequences from EMBL, and 16S rRNA sequences of bacteria from RDP, synthesized on the Agilent platform. The array was tested using 2 color dyes on cultured microbes and on clinical samples from sick and healthy people, looking for differences in clinically ill people compared to a number of healthy "controls".
