Project description:We explored transcriptional responses of the chytridiomycete Blastocladiella emersonii to some stress environmental conditions. Three distinct cDNA libraries constructed from mRNAs extracted from cells submitted to heat shock and different cadmium concentrations (50 and 100 mM) were sequenced and generated 6,350 high-quality EST sequences. These ESTs were assembled into 2,326 unigenes, being 51% of them not previously described in B. emersonii and consequently represent a worthy contribution of this work. We assigned to approximately 59% of the unigenes an orthologue in other organism, whereas 41% remained without a putative identification. Using the transcriptome data of B. emersonii we constructed a microarray slide containing 3,773 different cDNAs which comprise many biological processes of this fungus and analyzed its expression pattern in response to cadmium and heat shock. Through cDNA microarray assays we observed that 3,3% and 5,1% of the genes present in B. emersonii microarray slide were up-regulated in response to heat shock and cadmium, respectively. The main categories to which belong these genes were protein folding and proteolysis, proteins with antioxidant properties, cellular transport, carbohydrate and amino acid metabolism. Interestingly, in response to cadmium stress, B. emersonii induced six different genes codifying Glutathione S-transferases and six Metacaspases, as well as several proteins from sulfur amino acid metabolism. Keywords: stress response; heavy metal stress response; heat shock response
Project description:We performed shallow whole genome sequencing (WGS) on circulating free (cf)DNA extracted from plasma or cerebrospinal fluid (CSF), and shallow WGS on the tissue DNA extracted from the biopsy in order to evaluate the correlation between the two biomaterials. After library construction and sequencing (Hiseq3000 or Ion Proton), copy number variations were called with WisecondorX.
Project description:Due to its aquatic saprophytic lifestyle, the fungus Blastocladiella emersonii seems to be adapted to low aerated environments and thus can be an interesting model of study of the hypoxic stress response. Iron deprivation, a hypoxia-mimicking stimulus due to HIF-1 stabilization, and geldanamycin, that causes HIF-1 depletion by HSP90 inhibition, are helpful parameters to find an evidence of an oxygen-dependent regulatory mechanism in B. emersonii similar to the metazoan HIF-1 pathway.
Project description:Whole genome sequencing (WGS) of tongue cancer samples and cell line was performed to identify the fusion gene translocation breakpoint. WGS raw data was aligned to human reference genome (GRCh38.p12) using BWA-MEM (v0.7.17). The BAM files generated were further analysed using SvABA (v1.1.3) tool to identify translocation breakpoints. The translocation breakpoints were annotated using custom scripts, using the reference GENCODE GTF (v30). The fusion breakpoints identified in the SvABA analysis were additionally confirmed using MANTA tool (v1.6.0).
Project description:In principle, whole-genome sequencing (WGS) of the human genome even at low coverage offers higher resolution for genomic copy number variation (CNV) detection compared to array-based technologies, which is currently the first-tier approach in clinical cytogenetics. There are, however, obstacles in replacing array-based CNV detection with that of low-coverage WGS such as cost, turnaround time, and lack of systematic performance comparisons. With technological advances in WGS in terms of library preparation, instrument platforms, and data analysis algorithms, obstacles imposed by cost and turnaround time are fading. However, a systematic performance comparison between array and low-coverage WGS-based CNV detection has yet to be performed. Here, we compared the CNV detection capabilities between WGS (short-insert, 3kb-, and 5kb-mate-pair libraries) at 1X, 3X, and 5X coverages and standardly used high-resolution arrays in the genome of 1000-Genomes-Project CEU genome NA12878. CNV detection was performed using standard analysis methods, and the results were then compared to a list of Gold Standard NA12878 CNVs distilled from the 1000-Genomes Project. Overall, low-coverage WGS is able to detect drastically more (approximately 5 fold more on average) Gold Standard CNVs compared to arrays and is accompanied with fewer CNV calls without secondary validation. Furthermore, we also show that WGS (at ≥1X coverage) is able to detect all seven validated deletions larger than 100 kb in the NA12878 genome whereas only one of such deletions is detected in most arrays. Finally, we show that the much larger 15 Mbp Cri-du-chat deletion can be clearly seen at even 1X coverage from short-insert WGS.