Project description:Genome organization involves cis and trans chromosomal interactions, both implicated in gene regulation, development, and disease. Here, we focused on trans interactions in Drosophila, where homologous chromosomes are paired in somatic cells from embryogenesis through adulthood. We first addressed the long-standing question of whether pairing extends genome-wide and, to this end, developed a haplotype-resolved Hi-C approach that uses a new strategy to minimize homolog misassignment and thus robustly distinguish trans-homolog from cis contacts. This approach revealed striking genome-wide pairing in Drosophila embryos. Moreover, we discovered pairing to be surprisingly structured, with trans-homolog domains and interaction peaks, many coinciding with the positions of analogous cis features. We also found a significant correlation between pairing and the chromatin accessibility mediated by the pioneer factor Zelda. Our findings reveal a complex, highly structured organization underlying homolog pairing, first discovered more than a century ago. Overall design: This submission contains data from a Hi-C experiment on Early Drosophila embryos (2 biological replicates).
Project description:Influenza A(H1N1)pdm virus caused the first human pandemic of the 21st century. Although various probiotic Lactobacillus species have been shown to have anti-microbial effects against pneumonia-inducing pathogens, the prophylactic efficacy and mechanisms behind their protection remain largely unknown. Here, we evaluated the prophylactic efficacy of heat-killed Lactobacillus pentosus b240 against lethal influenza A(H1N1)pdm virus infection in a mouse model. To further define the protective responses induced by b240, we performed virologic, histopathologic, and transcriptomic analyses on the mouse lungs. Although we did not observe an appreciable effect of b240 on virus growth, cytokine production, or histopathology, gene expressional analysis revealed that oral administration of b240 differentially regulates antiviral gene expression in mouse lungs. Our results unveil the possible mechanisms behind the protection mediated by b240 against influenza virus infection and provide new insights into probiotic therapy. Six-week-old female BALB/c mice were used in the study. Oral administration of b240 was initiated in mice at six weeks of age. Mice were orally administered heat-killed Lactobacillus pentosus b240 every day at a dose of 10 mg/mouse in 200 μl of buffered saline for 5 weeks. The control group received saline. To investigate the effects of oral administration of b240 on host immune responses to CA04 virus infection, 9 mice per group were infected with 10 MLD50 of CA04 virus on day 21 post-b240 administration. Three mice per group were euthanized on days 1, 3, and 6 post-infection and their lungs were collected. To investigate the immune responses induced by oral administration of b240 in the lungs of uninfected mice, 15 mice per group were mock-infected with PBS on day 21 post-b240 administration. Three mice per group were euthanized on days 14, 21, 22, 24, and 27 post-b240 administration (-7, 0, 1, 3, and 6 days post-mock infection) and their lungs were collected. These lung tissues were subjected to microarray analysis (three biological replicates per each group).
Project description:Two winter wheat cultivars possessing a high tolerance to Al (Triticum aestivum L. cv. Atlas66 and the near isogenic line OK91G106, named Century-T in this work ) and two winter wheat cultivars with low tolerance to Al (T. aestivum L. cv. Bounty and the near isogenic line OK91G108, named Century-S in this work ) were grown as previously described, and treated under conditions where Al remains mostly under the Al3+ form . To reduce pH variations and ensure that Al speciation was stable throughout the experiment, at least 100 ml of solution was used for each plant. The root growth inhibition (RGI) is expressed as 100 × [1- (root growth of Al-treated seedling divided by the root growth of control seedlings)]. Root tips (5-10 mm) were isolated after 24 hours of exposure to Al and frozen on dry ice. Total RNA was isolated from the root tips (5 mm long) of 50 plants collected from the various genotypes using the RNeasy Plant Mini Kit (Qiagen). The RNA quality was assessed on agarose gels and with the Bioanalyzer 2100 (Agilent). Microarray profiling was performed according to Affymetrix protocols at the Functional Genomics Platform of the McGill University and Génome Québec Innovation Centre using the Affymetrix GeneChip Wheat Genome Array. Plants of the different lines were treated with Al concentrations resulting in 50% RGI: 50 ?M for the tolerant cultivars Atlas66 and Century-T and 5 ?M for the sensitive cultivars Bounty and Century-S. Three biological replicates were used and the results were analyzed using the Robust Multiarray Analysis (RMA version 0.2) freeware. Atlas66 and Bounty plants not exposed to Al were used as control for all the other conditions. Genes showing a differential expression greater than two-fold (RMA differential expression of log2 ? 1) between the Atlas66 and Bounty controls were excluded from the reference set as these may represent cultivar dependent differences in basal expression levels that could bias the analysis. Overall, this excluded 1.26% of the genes on the microarray (773 of 55,052) demonstrating that most genes are expressed at a similar level between the two cultivars. The microarray headings are the following : For each array headings: The first term represents the cultivar: A = Atlas66; B = Bounty, 106 = Century-T, 108 = Century-S. The second twerm represents the Al concentration: 0 = no aluminum 5 = 5 ?M aluminum and 50 = 50 ?M aluminum. The last term represents the sample replicate number S1, S2 or S3.
2008-06-30 | E-TABM-454 | ArrayExpress
Project description:Sixth Century Szólád and Collegno whole genomes
| PRJNA433631 | ENA
Project description:Sixth Century Szólád and Collegno 1240K capture
| PRJNA433632 | ENA
Project description:Resequencing Solanaceae (Potato and Tomato) 19th century samples
Project description:Sindbis virus is a mutagenic ssRNA arbor virus. The structural genome component of this virus can be exchanged for any transgene of < 6kb in size. Here we sequence transgenic elements inserted to the virus. Our EGFP transgene was analyzed as a function of time during viral replication to quantify the mutagenic rate of the virus. Our nanobody transgene was analyzed to assess cDNA library complexity and identity. Overall design: In this study we packaged transgenic virus carrying either EGFP or a library of nanobody cDNA sequences obtained from a 5HT2A receptor immunized llama. The EGFP packaged virus was applied to BHK21 (hamster) cells harboring the plasmid pCMV-SSG, a mammalian expression plasmid to drive constitutive production of the sindbis virus structural genome (SSG). This allows the virus to freely proliferate in the transfected culture. We collected virus from cell culture at 3, 6, 24, and 36 hours after infection and amplified the EGFP transgene. The amplicons were sequenced and sequence integrity of individual nucleotide positions was determined. The nanobody-containing Sindbis genome was directly amplified without culturing and sequenced. The resulting sequences were aligned to the FASTA files provided in this submission to determine if the FASTA deposited nanobody sequences were present in the originating viral titer.
Project description:Illinois protein lines (IPL) comprise lines known as Illinois high protein (IHP), Illinois low protein (ILP), Illinois reverse-low protein (IRLP) and Illinois reverse-high protein (IRHP) lines. These lines represent over a century old experiment containing lines that represent various levels of nitrogen metabolism potential as a function of their grain protein concentration. Microarray experiment was conducted using 16 DAP seeds from the above mentioned four IPLs. Keywords: Genotype response Overall design: Technical replicates, dye swaps, technical replicates of dye swaps, no biological replicates
Project description:Food and diet were class markers in nineteenth-century Ireland, which became evident as nearly one million people, primarily the poor and destitute, died as a consequence of the notorious Great Famine of 1845–1852. Famine took hold after a blight (Phytophthora infestans) destroyed the only means of subsistence—the potato crop—for a significant proportion of the population. This study seeks to elucidate the variability of diet in mid-nineteenth-century Ireland through microparticle and proteomic analysis of human dental calculus samples (n = 42) from victims of the Famine. The sample derives from remains of people who died between August 1847 and March 1851 while receiving poor relief as inmates in the union workhouse in the city of Kilkenny (52°39’N, -7°15’W). The results corroborate the historical accounts of food provisions before and during the Famine, with evidence of corn (maize), potato and cereal starch granules from the microparticle analysis and milk proteins from the proteomic analysis. Unexpectedly, there is also evidence of egg proteins—a food source generally reserved only for export and the better-off social classes—which highlights the variability of the Famine experience for those who died. Through historical contextualisation, this study shows how the notoriously monotonous potato diet of the nineteenth-century Irish poor was supplemented by other foodstuffs on an opportunistic basis. While the Great Irish Famine was one of the worst subsistence crises in history, it was foremost a social disaster induced by the lack of access to food and not the lack of food availability.
Project description:Long non-coding RNAs (lncRNAs) are a new arm of gene regulatory mechanism as discovered by sequencing techniques and follow-up functional studies. There are only few studies on lncRNAs as related to gene expression regulation and anti-viral activity during influenza virus infection. We sought to identify and characterize lncRNAs involved in influenza virus replication. In the current study, we identified dys-regulated lncRNAs in influenza virus-infected human lung epithelial A549 cells using RNA sequencing in A549 cells. Overall design: RNA sequencing of A549 cells with Influenza virus and MOCK infection.