Project description:We performed whole-transcriptome RNA sequencing (RNA-Seq) to compare transcript abundances in the skin of DENV- or ZIKV-infected AG6 mice to those in the skin of uninfected mice.
Project description:Crab is one of the major source for V. parahaemolyticus outbreak among aquatic products in Northeast Asian due to improperly cooking and wound infection by mishandling. However, there is no report on whole genome sequence of V. parahaemolyticus isolated from contaminated crab, thus no information is available for major virulence factors about V. parahaemolyticus obtained from crab. Therefore, the analysis of transcriptome of isolated V. parahaemolyticus from crab products are necessary to investigate potential risk of foodborne illness by contaminated products.
Project description:A holistic approach to examine differential gene expression profiles of transcripts relevant to the moulting process, across all moult cycle stages, was used in this study. Custom cDNA microarrays were constructed for P. pelagicus. The printed arrays contained 5000 transcripts derived from both the whole organism, and from individual organs such as the brain, eyestalk, mandibular organ and Y-organ from all moult cycle stages. A total of 556 clones were sequenced from the cDNA libraries used to construct the arrays. These cDNAs represented 217 singlets and 62 contigs, resulting in 279 unique putative genes. Moult cycle-related differential expression patterns were observed for many transcripts. Keywords: cyclic moult stage comparison of the crab Portunus pelagicus
2008-01-03 | GSE6997 | GEO
Project description:Identifying neuropeptides and their GPCRs in Central Nervous System Transcriptome of swimming crab, Portunus trituberculatus.
Project description:Purpose:The mud crab Scylla paramamosain is an economically important marine crab in China suffering from severe outbreaks of infectious disease caused by marine bacteria such as Vibrio Parahaemolyticus, resulting in great economic losses. However, the mechanisms involved in the immune response of this crab to bacterial infection are not fully understood. To understand the molecular mechanisms underlying the immune response to such pathogenic bacteria, we used high-throughput deep sequencing technology to investigate the transcriptome and comparative expression profiles of the mud crab S.paramamosain infected with V.parahaemolyticus. Methods: The hemocytes sampled at 0-24h after infection with V.parahaemolyticus were used for transcriptome analysis. The hemocytes sampled at 24 h after injections with V.parahaemolyticus and no injected 0h(as control) were used for gene expression profiling analysis. Results: A total of 52,934,042 reads were obtained and assembled into 186,193 contigs in transcriptional responses of the V.parahaemolyticus-infected mud crab. Via annotation to the NCBI database and the Swissprot database, we obtained 48,934 identified unigenes. In total, 10,139(20.7%) unigenes were classified into Gene Ontology, and 25,349 unigenes were found in 20 KEGG categories. These genes included representatives from almost all functional categories. By using Solexa/Illumina's DeepSAGE, 1213 differentially expressed genes (P value < 0.05) were detected in comparative analysis of the expression profiles between V.parahaemolyticus-infected crabs and control crabs, including 538 remarkably upregulated genes and 675 remarkably downregulated genes. Conclusions: Based on our results, we conclude that the inflammatory response may play an important role in the early stages of infection. The signaling cascades such as the chemokine, JAK-STAT, and MAPK pathways are regulated by V.parahaemolyticus infection. These results revealed changes of multiple signaling pathways involved in immunity during V.parahaemolyticus infection, which will facilitate our comprehensive understanding of the mechanisms involved in the immune response to bacterial infection in the mud crab.
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular pathways. The objective of this study was to compare transcriptome analysis (RNA-seq) with microarray and quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) for HIV-1-infected and uninfected MDMs and to evaluate the optimal protocol for high-throughput data analysis Methods: The mRNA profiles of HIV-1-infected and uninfected MDMs were generated by deep sequencing, in triplicate, using Illumina GAIIx. The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays. Results: Using an optimized data analysis workflow, we mapped about 20 million sequence reads per sample to the human genome and identified 60505 transcripts in the HIV-1-infected or uninfected MDMs. RNA-seq data confirmed stable expression of 15 known housekeeping genes, and 10 of these were validated with qRT–PCR. RNA-seq data had a linear relationship with qRT–PCR for more than four orders of magnitude and a goodness of fit (R2) of 0.8953. Approximately 10% of the transcripts showed differential expression between the HIV-1-infected or uninfected MDMs, with a fold change ≥1.5 and p value <0.05. Altered expression of 48 genes was confirmed with qRT–PCR, demonstrating the high degree of sensitivity of the RNA-seq method. Conclusions: Our study represents the detailed analysis of HIV-1-infected macrophages transcriptomes, with biologic replicates, generated by RNA-seq technology. Our results show that NGS offers a comprehensive and more accurate quantitative and qualitative evaluation of mRNA content within a cell or tissue. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions.