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109 results for “DNA sequence analysis”
Transcriptome analysis of the effect of over-expressing H2A.J mutants in proliferating WI38 fibroblasts for the paper entitled: The H2A.J histone variant contributes to Interferon-Stimulated Gene expression in senescence by its weak interaction with H1 and the derepression of repeated DNA sequences
<p>Abstract for overall study:</p> <p>The histone variant H2A.J was previously shown to accumulate in senescent human fibroblasts with persistent DNA damage to promote inflammatory gene expression, but its mechanism of action was unknown. We show that H2A.J accumulation contributes to weakening the association of histone H1 to chromatin and increasing its turnover. Decreased H1 in senescence is correlated with increased expression of some repeated DNA sequences, increased expression of STAT/IRF transcription factors, and transcriptional activation of Interferon-Stimulated Genes (ISGs). The H2A.J-specific Val-11 moderates the transcriptional activity of H2A.J, and H2A.J-specific Ser-123 can be phosphorylated in response to DNA damage with potentiation of its transcriptional activity by the phospho-mimetic S123E mutation. Our work demonstrates the functional importance of H2A.J-specific residues and potential mechanisms for its function in promoting inflammatory gene expression in senescence.</p> <p>Specific description for this dataset:</p> <p>H2A.J differs from canonical H2A only by a valine at position 11 instead of alanine, and the 7 C-terminal amino acids containing a potential minimal phosphorylation site SQ for DNA-damage response kinases. To test the functional importance of these H2A.J-specific sequences, we mutated Val-11 to Ala as is found in all canonical H2A sequences, and we mutated Ser-123 to either Glu to mimic a phospho-serine residue or to Ala to prevent phosphorylation. We also substituted the C-terminus of H2A.J with the C-terminus of H2A. These mutants, WT-H2A.J and canonical H2A-type1 were ectopically expressed in proliferating fibroblasts, and their microarray transcriptomes were compared to that of proliferating and senescent fibroblasts without ectopic histone expression. Genome-wide transcriptome analysis indicated that senescent fibroblasts clustered distinctly from proliferating fibroblasts, and proliferating fibroblasts expressing the H2A.J-V11A and H2A.J-S123E mutants clustered distinctly from fibroblasts expressing the other H2A.J mutants, WT-H2A.J, and H2A. Hallmark gene set enrichment analysis of the transcriptomes of fibroblasts expressing H2A.J-V11A or H2A.J-S123E versus control proliferating fibroblasts indicated that they showed the same highly significant enrichment for the Epithelial-Mesenchyme Transition, TNF-Alpha Signaling Via NF-kB, and Inflammatory Response gene sets. Notable inflammatory genes including IL1A, IL1B, IL6, CXCL8, and CCL2 are contained in these gene sets and are often induced in senescence as part of the senescence-associated secretory phenotype. Heat maps showed that the H2A.J-V11A and H2A.J-S123E mutants were particularly apt at activating the expression of these inflammatory genes in proliferating fibroblasts</p>
Figure 3 in Diceratocephala boschmai (Platyhelminthes: Temnocephalida) from crayfish farms in Thailand: investigation of the topographic surface and analysis of 18S ribosomal DNA sequences
Figure 3. The neighbor-joining phylogenetic tree based on the 18S rDNA gene, showing the relationships of D. boschmai with 29 other turbellarian species.
Figure 2 in Diceratocephala boschmai (Platyhelminthes: Temnocephalida) from crayfish farms in Thailand: investigation of the topographic surface and analysis of 18S ribosomal DNA sequences
Figure 2. Surface topography of D. boschmai. A–C) Unhatched and hatched eggs; D) ventral view of a specimen; E) mouth with protruding pharynx; F) thread-like filaments adhered to the pharynx; G, H) a higher magnification of write-dot boxes in 2E; I, J) a higher magnification of write-dot boxes in 2D; K) ventral view of a specimen at posterior end; L) dorsal view of a specimen; M) a higher magnification of write-dot box in Figure L. ad, adhesive disc; af, adhered filaments; ci, ciliated cell; ds, double spines; fi, filament; gr, groove; gv, gravel-like units; in, intertentacular flange; mo, mouth; op, opercular plate; pe, peduncle; pi, pit; sp, single spine; tb, trabecular meshwork; tc, tentacle; th, thread-like filaments; tr, trunk; vi, villi..
Figure 1. A in Diceratocephala boschmai (Platyhelminthes: Temnocephalida) from crayfish farms in Thailand: investigation of the topographic surface and analysis of 18S ribosomal DNA sequences
Figure 1. A) C. destructor harboring adult D. boschmai and eggs of flatworm; B) dorsal view of an extending body; C) diagram of organ structures in dorsal view; D) diagram of reproductive complex; E, F) photomicrograph and diagram of penial stylet, respectively; G) unhatched and hatched eggs. ad, adhesive disc; at, atrium; cv, contractile vesicle; ds, dorsal side; es, ejaculatory sac; ey, eye; ev, excretory vesicle; fi, filament; in, intertentacular flange; ine, intestine; int, introvert; mo, mount; ov, ovary; pe, peduncle; pf, plane of fracture; ph, pharynx; pn, subepidermal pigment network; ps, penial stylet; rv, resorbens vesicle; s, stalk; se, seminal vesicle; sr, seminal receptacle; sp, sclerotized papillae; tc, tentacle; te, testis; tg, tentacular gland; ve, vasa efferentia; vg, vagina; vi, vitellaria; vs, ventral side.
Text-fig. 4. Electrophoresis after amplification: Electrophoretical analysis of mitochondrial DNA. mtDNA sequences were amplified by primers F15.412 and R16.169 (450 bp), R16.269 (550 bp), R16.519 (800 bp). Lane 1 are primers F15.412 + R16.169, lane 2 primers F15.412 + R16.269, lane 3 primers F15.412 + R16.519, NC – negative control – water, L – 100 bp DNA ladder (band size from 100 bp to 1500 bp). in Genetic Analysis Of Possibly The Oldest Greyhound Remains Within The Territory Of The Czech Republic As Proof Of A Local Elite Presence At Chotěbuz-Podobora Hillfort In The 8 -9 Century Ad
Text-fig. 4. Electrophoresis after amplification: Electrophoretical analysis of mitochondrial DNA. mtDNA sequences were amplified by primers F15.412 and R16.169 (450 bp), R16.269 (550 bp), R16.519 (800 bp). Lane 1 are primers F15.412 + R16.169, lane 2 primers F15.412 + R16.269, lane 3 primers F15.412 + R16.519, NC – negative control – water, L – 100 bp DNA ladder (band size from 100 bp to 1500 bp).
Fig 1 in Evolutionary relationships of Macaca fascicularis fascicularis (Raffles 1821) (Primates: Cercopithecidae) from Singapore revealed by Bayesian analysis of mitochondrial DNA sequences
Fig 1. Map of Southeast Asia showing the approximate location of the new (Singapore and Bali) and GenBank sequences included in the study. Numbers correspond to the following locations (haplotype IDs in parentheses): ★, Singapore (Sing1–3); 1, Vietnam (Viet1 & 2); 2, Cambodia (Camb1 & 2); 3, Thailand (Thai1); 4, Thailand (Thai2); 5, Malaysia (Selangor1 & 2); 6, Malaysia (Johor); 7, south Sumatra, Indonesia (Sumatra1 & 2, Java1); 8, Java (Java1); 9, Kalimantan, Borneo (Borneo3); 10, Sarawak, Borneo (Borneo1); 11, Sepilok, Borneo (Borneo2); 12, Bali, Indonesia (Bali1 & 2); 13, Sibuyan, Philippines (Phil1); 14, Bangkok, Thailand (Thai3 & 4); 15, Malaysia (W. Malay); 16, Malaysia (E. Malay2); 17 Malaysia (E. Malay1);18, north Sumatra (Sumatra3–6, 9); 19, west Borneo, Indonesia (Borneo9); 20, west Borneo, Indonesia (Borneo4–7); 21, central Borneo, Indonesia (Borneo4 & 6); 22, Bangka, south Sumatra (Sumatra 7 & 8); 23, Java, Indonesia (Java2 & 3); 24, northeast Borneo, Indonesia (Borneo8); 25, Mindanao, Philippines (Phil2); 26, Timor (Timor). Several Borneo haplotypes appear in multiple locations.
Fig 4. Median-joining haplotype network for M in Evolutionary relationships of Macaca fascicularis fascicularis (Raffles 1821) (Primates: Cercopithecidae) from Singapore revealed by Bayesian analysis of mitochondrial DNA sequences
Fig 4. Median-joining haplotype network for M. fascicularis. The size of the circular nodes representing haplotypes is proportional to the number of sequences comprising the haplotype. Shading of circular nodes corresponds to general geographic groupings including Sundaic islands (white), mainland Indochina (gray), Malay Peninsula and northern Sumatra (dark gray), and Singapore (black). Haplotype identifications are presented in Table 1.
Fig 3 in Evolutionary relationships of Macaca fascicularis fascicularis (Raffles 1821) (Primates: Cercopithecidae) from Singapore revealed by Bayesian analysis of mitochondrial DNA sequences
Fig 3. Phylogenetic tree topology from Bayesian inference of 12S/tRNA-val/16S mtDNA sequences using a Birth-Death speciation tree prior, and HYK+G+I nucleotide substitution model in BEAST v2.1.3. Lettered identifications for clades are presented below the branches at major nodes. Posterior probabilities are displayed above the branches at nodes. Numbers in parentheses appearing with haplotype identifications are presented in Table 1, and correspond to numbered locations presented on the Figure 1 map. The Singapore haplotypes form two phylogenetic subgroupings, one from the Bukit Timah Nature Reserve (Sing1) and the other from the Central Catchment Nature Reserve (Sing2 & Sing3).
Fig 2 in Evolutionary relationships of Macaca fascicularis fascicularis (Raffles 1821) (Primates: Cercopithecidae) from Singapore revealed by Bayesian analysis of mitochondrial DNA sequences
Fig 2. Map of central Singapore showing the sampling locations in the Bukit Timah (BTNR) and Central Catchment (CCNR) Nature Reserves. Map created using ArcGIS® (ESRI® 2015).
Fig. 1 in Populations analysis of the Brazilian Sharpnose Shark Rhizoprionodon lalandii (Chondrichthyes: Carcharhinidae) on the São Paulo coast, Southern Brazil: inferences from mt DNA sequences
Fig. 1. Median-joining haplotype network. The haplotypes are represented by circles, with the width proportional to their frequencies. Black circles correspond to Praia Grande, white to Ubatuba, and gray to Itanhaém samples. Each branch corresponds to a single mutation, except line a (with 2 mutations) and line b (with 3 mutations).
Multimodal Epigenetic Sequencing Analysis (MESA) of Cell-free DNA for Non-invasive Colorectal Cancer Detection
<p>Processed data (feature-by-sample matrices) of non-disruptive bisulfite-free methylation sequencing for cfDNA samples from 4 clinical cohorts (Cohort 1, Cohort 2, Cohort 3, and cfTAPS dataset). Codes used to repeat the results in our paper can be found https://rpubs.com/LiYumei/926228 and https://github.com/ChaorongC/MESA. </p>
Comprehensive, targeted eukaryotic metagenomics analysis of environmental DNA biodiversity using Oxford Nanopore sequencing
<p><span>Metagenomics has become a prominent technology for studying the functional potential of all organisms in a microbial and eukaryotic community. The study of symbiotic organisms from different classes or kingdoms, including those previously unknown, is possible with simultaneous and equally efficient metagenomic analysis of these species. A variety of targeted primer sets are used for eukaryotic metagenomic biodiversity, including those that are universal for specific families, classes</span><span>,<span> or kingdoms. The most universal for all existing cellular organisms is the presence of ribosomal RNA encoding gene sequences. For eukaryotic sequences, these are 16S and 23s rDNA, </span>and <span>for eukaryotic sequences of nuclear (18S and 28S) and mitochondrial (12S and 16S) ribosomal RNA. Here we present the application of the eukaryotic metagenomics approach to the simultaneous, quantitative</span>,<span> and unbiased identification of most eukaryotic species. To achieve this, we have developed a universal PCR assay that targets the most conservative nuclear regions of the ribosomal gene for all cellular organisms, including plants, algae, fungi, protists, insects</span>,<span> and animals. The amplification product contains polymorphic regions of both ribosomal genes and the intergenic spacer. The size of the PCR products varies by class, kingdom</span>,<span> or domain, ranging from 2 kb for fungi to 7 kb for birds. This assay is also adapted for use with the Oxford Nanopore Rapid Barcoding Library Kit, which enables metagenomic biodiversity analysis. Our approach provides a rapid, sensitive</span>,<span> and equally efficient way to study the composition of eDNA from mixed species in the environment. This protocol reduces the time and cost of metagenomic biodiversity analysis using Oxford Nanopore sequencing. We can efficiently analyze the biodiversity of mixed species present in environmental samples.</span></span></p>
Oxford Nanopore sequencing for comprehensive, targeted eukaryotic metagenomics analysis of environmental DNA biodiversity
<p><span>The study of symbiotic organisms from different classes or kingdoms, including those previously unknown, is possible with simultaneous and equally efficient metagenomic analysis of these species. A variety of targeted primer sets are used for eukaryotic metagenomic biodiversity, including those that are universal for specific families, classes</span><span>,<span> or kingdoms. The most universal for all existing cellular organisms is the presence of ribosomal RNA encoding gene sequences. For eukaryotic sequences, these are 16S and 23s rDNA, </span>and <span>for eukaryotic sequences of nuclear (18S and 28S) and mitochondrial (12S and 16S) ribosomal RNA. Here, we present the application of the eukaryotic metagenomics approach to the simultaneous, quantitative</span>,<span> and unbiased identification of most eukaryotic species. </span></span></p>
DNA metabarcoding sequence data for diet analysis of caribou
<p>Woodland caribou (<em>Rangifer tarandus caribou</em>) are threatened in Canada due to the drastic decline in population size caused primarily by human-induced landscape changes that decrease habitat and increase predation risk. Conservation efforts have largely focused on reducing predators and protecting critical habitat, whereas research on dietary niches and the role of potential food constraints in lichen-poor environments is limited. To improve our understanding of dietary niche variability, we used a next-generation sequencing approach with metabarcoding of DNA extracted from faecal pellets of woodland caribou located on Lake Superior in lichen-rich (mainland) and lichen-poor (island) environments. Amplicon sequencing of fungal ITS2 region revealed lichen-associated fungi as predominant in samples from both populations, but amplification at the chloroplast <em>trnL </em>region, which was only successful on island samples, revealed primary consumption of yew based on relative read abundance (<em>Taxus spp.</em>; 83.68%) with dogwood (<em>Cornus spp</em>.; 9.67%) and maple (<em>Acer spp.</em>; 4.10%) also prevalent. These results suggest that conservation efforts for caribou need to consider the availability of food resources beyond lichen to ensure successful outcomes. More broadly, we provide a reliable methodology for assessing ungulate diet from archived faecal pellets that could reveal important dietary shifts over time in response to climate change.</p>
Data for paper: Analysis and control of untemplated DNA polymerase activity for guided synthesis of kilobase-scale DNA sequences
<p>This deposit contains data for the paper entitled: "<strong>Analysis and control of untemplated DNA polymerase activity for guided synthesis of kilobase-scale DNA sequences</strong>"</p> <p>Contents include:</p> <ul> <li><strong>00_sequencing-metadata.xlsx</strong> - Metadata for each sequencing sample.</li> <li><strong>01_raw-fastqs.zip</strong> - Raw basecalled FASTQ data.</li> <li><strong>02_clean-fastas.zip</strong> - Cleaned FASTA files (removal of adapters and barcode sequences).</li> <li><strong>03_read-statistics.zip</strong> - Read statistics for all samples.</li> <li><strong>04_sequence-analysis.zip</strong> - Sequence analysis output for all samples.</li> <li><strong>05_qpcr-data.xlsx</strong> - qPCR data for all the dNTP mixes studied.</li> <li><strong>06_analysis-scripts.zip</strong> - Analysis scripts.</li> </ul>
Data from: Adaptive radiation of the Callicarpa genus in the Bonin Islands revealed through double-digest restriction site–associated DNA sequencing analysis
Open the record for dataset details and reuse information.
DNA metabarcoding sequence data for diet analysis of caribou
Open the record for dataset details and reuse information.
Transcriptome analysis of WT versus H2A.J-KO MEFs for the paper entitled: The H2A.J histone variant contributes to Interferon-Stimulated Gene expression in senescence by its weak interaction with H1 and the derepression of repeated DNA sequences
<p>Abstract for overall study:</p> <p>The histone variant H2A.J was previously shown to accumulate in senescent human fibroblasts with persistent DNA damage to promote inflammatory gene expression, but its mechanism of action was unknown. We show that H2A.J accumulation contributes to weakening the association of histone H1 to chromatin and increasing its turnover. Decreased H1 in senescence is correlated with increased expression of some repeated DNA sequences, increased expression of STAT/IRF transcription factors, and transcriptional activation of Interferon-Stimulated Genes (ISGs). The H2A.J-specific Val-11 moderates the transcriptional activity of H2A.J, and H2A.J-specific Ser-123 can be phosphorylated in response to DNA damage with potentiation of its transcriptional activity by the phospho-mimetic S123E mutation. Our work demonstrates the functional importance of H2A.J-specific residues and potential mechanisms for its function in promoting inflammatory gene expression in senescence.</p> <p>Specific description for this dataset:</p> <p>We further tested a role for H2A.J in Interferon-Stimulated Gene expression by analyzing the transcriptome of WT and H2A.J MEFs induced into senescence by etoposide. TruSeq stranded DNA libraries were prepared from polyA-selected RNA and sequenced as 43 bp paired-end reads. The fastq sequences were mapped to Gencode.vM24.transcripts.fa.gz (GRCm38 transcriptome) with salmon. Read counts were then aggregated to the gene level with tximeta, and differential gene expression was analysed with DESeq2, edgeR, and limma-voom. Gene set enrichment analysis was performed with camera.</p> <p>The transciptomes of senescent WT and H2AFJ-KO showed strong separation from proliferating MEFs, and a weaker separation distinguished WT and H2A.J-KO MEFs. Strikingly, gene set enrichment analysis indicated highly significant defects in Interferon Response Gene Expression in the H2A.J-KO MEFs in senescence with significant down-regulation in senescent H2A.J-KO cells of a series of oligoadenylate synthase genes (Oas1g, Oas1a, Oasl1, Oas2, Oasl2) and several ISGs. Thus, H2A.J also contributes to ISG expression in the heterologous context of senescent MEFs.</p>
Data from: Estimation of a killer whale (Orcinus orca) population's diet using sequencing analysis of DNA from feces
Estimating diet composition is important for understanding interactions between predators and prey and thus illuminating ecosystem function. The diet of many species, however, is difficult to observe directly. Genetic analysis of fecal material collected in the field is therefore a useful tool for gaining insight into wild animal diets. In this study, we used high-throughput DNA sequencing to quantitatively estimate the diet composition of an endangered population of wild killer whales (Orcinus orca) in their summer range in the Salish Sea. We combined 175 fecal samples collected between May and September from five years between 2006 and 2011 into 13 sample groups. Two known DNA composition control groups were also created. Each group was sequenced at a ~330bp segment of the 16s gene in the mitochondrial genome using an Illumina MiSeq sequencing system. After several quality controls steps, 4,987,107 individual sequences were aligned to a custom sequence database containing 19 potential fish prey species and the most likely species of each fecal-derived sequence was determined. Based on these alignments, salmonids made up >98.6% of the total sequences and thus of the inferred diet. Of the six salmonid species, Chinook salmon made up 79.5% of the sequences, followed by coho salmon (15%). Over all years, a clear pattern emerged with Chinook salmon dominating the estimated diet early in the summer, and coho salmon contributing an average of >40% of the diet in late summer. Sockeye salmon appeared to be occasionally important, at >18% in some sample groups. Non-salmonids were rarely observed. Our results are consistent with earlier results based on surface prey remains, and confirm the importance of Chinook salmon in this population's summer diet.
APPENDIX. GenBank accession numbers of all DNA sequences of Cophyla used in this study. NA, not applicable. Asterisks mark cases where sequences from different samples were combined to chimeric terminals for analysis. in Description of the lucky Cophyla (Microhylidae, Cophylinae), a new arboreal frog from Marojejy National Park in north-eastern Madagascar
APPENDIX. GenBank accession numbers of all DNA sequences of Cophyla used in this study. NA, not applicable. Asterisks mark cases where sequences from different samples were combined to chimeric terminals for analysis.
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