Project description:Entamoeba histolytica is the parasite causing amoebiasis, an infectious disease targeting the intestine and liver of humans. The molecules involved in amoeba adaptation to different iron conditions during the invasive process are unknown. To investigate the effects of iron availability on gene expression in E. histolytica, we determined by microarray experiments the gene expression profile of parasites grown in the presence of different iron concentrations. Conditions included low amounts of iron, iron starvation and iron starvation supplemented with hemoglobin (Hb). Genes encoding important proteins involved in iron metabolism, reported in other organism such as bacteria, were identified for the first time in E. histolytica. The influence of iron on the amount of these transcripts was confirmed by quantitative real-time PCR and their protein products were analyzed by Western blots
Project description:Entamoeba histolytica is a protozoan parasite which causes colitis and liver abscesses. A pilot microarray consisting of 360 unique parasite genes was constructed using identical methods to the larger array (1,971 unique genes). The four arrays in this data set were used to ascertain whether the microarrays would be useful in detecting changes in transcript abundance by exposing parasites to heat shock (42 0C for 1 hr). Approximately, 17% of the genes were regulated by at least two fold including many genes previously shown to be involved in heat shock response. This data confirmed that the genomic DNA arrays were useful in detecting changes in transcript abundance. Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: Logical Set
Project description:Little is known about the extent of genetic variability among Entamoeba strains and potential genotypic associations with virulence. Variable phenotypes have been identified for Entamoeba strains. E. histolytica is invasive and causes colitis and liver abscesses, but only in 10% of infected individuals; 90% of subjects remain asymptomatically colonized. E. dispar, a closely related species, appears to be incapable of causing invasive disease. In order to determine the extent of genetic diversity among Entamoeba strains we have developed an E. histolytica genomic DNA microarray and used it to genotype strains of E. dispar and E. histolytica. Based on the identification of divergent genetic loci, all six strains (four EH and two ED) had unique genetic fingerprints. Genomic regions with unusually high levels of divergence were identified indicating that structural or evolutionary pressures are molding selective regions of the Entamoeba genome. Comparison of divergent genetic regions allowed us to readily distinguish between EH and ED, identify novel genetic regions that may be used for strain and species typing, and identity a number of novel potential virulence determinants. Among these are Androgen Inducible Gene1, a CXXC receptor kinase, a peroxiredoxin 1-related gene, a Ras family member gene, a Rab geranylgeranyltransferase, and a gene with a UPF0034 domain. Among the four EH strains, an avirulent strain EH (Rahman) was the most divergent and phylogenetically distinct raising the intriguing possibility that genetic subtypes of E. histolytica may be at least partially responsible for the observed variability in clinical outcomes. Our approach shows the utility of a microarray-based genotyping assay to identify genetic variability between Entamoeba isolates and can readily be applied to the study of clinical isolates. A genotyping experiment design type classifies an individual or group of individuals on the basis of alleles, haplotypes, SNP's. Keywords: genotyping_design
Project description:Entamoeba histolytica is a protozoan parasite which causes colitis and liver abscesses. Using a genomic DNA microarray consisting of 1.6 - 2.0 kb genomic inserts we have generated a transcriptional profile of 1,971 unique parasite transcripts. The arrays in this experiment set were used to (1) estimate relative transcript abundance for Entamoeba histolytica (HM-1:IMSS) trophozoites in mid-logarithmic growth and (2) to examine changes in the transcriptional profile of Entamoeba histolytica when the parasite interacts with colonic epithelial cells (Caco-2). A time course was used such that RNA was isolated from ameba alone and ameba + Caco-2 cells at 3hrs, 6hrs and 9hrs corresponding to 10%, 50% and 90% cell monolayer destruction. At least two biological experiments and three replicates were used for each time point. Groups of assays that are related as part of a time series. Keywords: time_series_design