Transcription profiling by high throughput sequencing of Rana chensinensis oviduct from the initial growth period for de novo gene expression analysis
ABSTRACT: To obtain an overview of the Oviductus Ranae gene expression profile during initial growing period, a cDNA sample was prepared from Oviductus Ranae and sequenced using the Illumina sequencing platform
Project description:In order to analyze the transcriptome of ginseng root during leaf-expansion period and discover the genes during development, a cDNA sample was prepared from the leaf-expansion period of ginseng root and sequenced using the Illumina sequencing platform.The transcriptomic sequencing technology was set up the first time for five years the transcription of the ginseng root in the leaf-expansion period.
Project description:In order to research the ginseng leaf-stem gene expression profiles of and dig out its function genes in the leaf-expansion period, the transcriptomic sequencing technology was set up the first time for five years the transcription of the Panax ginseng leaf-stem in the leaf-expansion period.
Project description:Panax ginseng C.A. Meyer is one of the most popular medicinal herbs. In order to research the genes that related to the flowering period of ginseng, and find out the antifungal proteins and transcription factors that combat various biotic and abiotic stress, a cDNA sample was prepared from the flowering period ginseng root of a five-year-old plant and sequenced using the Illumina sequencing platform. In this study, we produced nearly 40 million sequencing reads. These reads were assembled into 134,045 contigs using Trinity software (mean size: 282 bp). Based on a similarity search with known proteins, we identified 79,307 sequences with a cut-off E-value of 10-5. Assembled sequences were then annotated using gene ontology (GO) terms, clusters of orthologous group (COG) classifications and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways respectively.
Project description:Background: Current evidence indicates that even low-level lead (Pb) exposure can have detrimental effects, especially in children. We tested the hypothesis that Pb exposure alters gene expression patterns in peripheral blood cells and that these changes reflect dose-specific alterations in the activity of particular pathways. Methodology/Principal Finding: Using Affymetrix Mouse Genome 430 2.0 arrays, we examined gene expression changes in the peripheral blood of female Balb/c mice following exposure to per os lead acetate trihydrate or plain drinking water for two weeks and after a two-week recovery period. Data sets were RMA-normalized and dose-specific signatures were generated using established methods of supervised classification and binary regression. Pathway activity was analyzed using the ScoreSignatures module from GenePattern. Conclusions/Significance: The low-level Pb signature was 93% sensitive and 100% specific in classifying samples a leave-one-out crossvalidation. The high-level Pb signature demonstrated 100% sensitivity and specificity in the leave-one-out crossvalidation. These two signatures exhibited dose-specificity in their ability to predict Pb exposure and had little overlap in terms of constituent genes. The signatures also seemed to reflect current levels of Pb exposure rather than past exposure. Finally, the two doses showed differential activation of cellular pathways. Low-level Pb exposure increased activity of the interferon-gamma pathway, whereas high-level Pb exposure increased activity of the E2F1 pathway. We isolate total RNA from 72 mouse whole blood samples. These included samples following a 2-week exposure to lead acetate trihydrate (untreated controls = 7; Low Pb 5ug/mL drinking water = 15; High Pb 50ug/mL drinking water = 15) and additional samples following a 2-week recovery period with plain drinking water (untreated controls = 7; Low Pb group = 15; High Pb group = 13).
Project description:Abiotic stress is a major factor affecting the growth, development, yield and quality in plants including the important biomass yield such as in a bioenergy feedstock crops. In a first of its kind, RNA-seq based high resolution survey of abiotic stress-induced transcriptome of bioenergy feedstock model plant western poplar (Populus trichocarpa accesion Nisqually-1) was carried out by way of 81 libraries made from total RNA isolated from three tissues, mature vascular leaf, stem xylem and root sampled from plants subjected to untreated control and treated with four individual stress treatments cold, heat, drought and high salinity. For every tissue and treatment type including controls, three individual plants were used per treatment as biological replicates. This research project is supported by the DOE Office of Science, Office of Biological and Environmental Research (BER), USA, grant no. DE-SC0008570
Project description:PURPOSE Diseases that involve choroidal or retinal endothelial vascular cells are leading causes of vision loss: age-related macular degeneration, retinal ischemic vasculopathies and non-infectious posterior uveitis. Proteins differentially expressed by these endothelial cell populations are potential drug targets. We used deep proteomics to define the molecular phenotype of human choroidal and retinal endothelial cells at the protein level. METHODS Choroidal and retinal endothelial cells were separately isolated from 5 human eye pairs by selection on CD31. Total protein was extracted and digested, and peptide fractions were analysed by reverse-phase liquid chromatography tandem mass spectrometry. Peptide sequences were assigned to fragment ion spectra and proteins were inferred using public protein databases. Protein abundance was determined by spectral counting. Protein expression in choroidal versus retinal endothelial cells was compared using edgeR package in R, and annotation enrichment analysis was performed. RESULTS Human choroidal or retinal endothelial cells expressed 5042 non-redundant proteins. Setting the false discovery rate at 5%, 498 proteins (14.4%) of 3454 quantifiable proteins with minimum mean spectral counts of 2.5 were differentially expressed between cell populations. Choroidal and retinal endothelial cells were enriched in angiogenic proteins, and retinal endothelial cells were also enriched in immunologic proteins. CONCLUSIONS This work demonstrates the protein heterogeneity of human choroidal and retinal vascular endothelial cells and provides multiple candidates for further study as novel treatments or drug targets for posterior eye diseases.
Project description:We conducted micro-array analysis to quantify the global transcriptome variations in floral organs of a male and female tree allowing for identification of sex-linked transcripts. We used RNA samples from male floral buds in August and female floral buds in September. Bud scale were removed. While the sampling time differed, the developmental stage of the floral organs was similar between the male and female. Five independent samples of floral bud tissues with bud scales removes were collected from the upper crown of a sexually mature male tree and female tree. RNA was extracted from tissues and hybridized on Affymetrix Genechip Poplar Genome Array.
Project description:This experiment was to look at the change in gene expression in oral epithelial cells infected for 6 h or 24 h with Candida albicans. The intent was to determine what changes were driven by early and late recognition of wild-type. invasive Candida albicans
Project description:Russeting of apple fruit is a non-invasive physiological disorder. It occurs mainly in 'Golden Delicious' apple and its hybrids, while understanding of its molecular mechanism is still limited. In this study, we used mRNA sequencing and an isobaric tag for relative and absolute quantitation-based quantitative (iTRAQ) proteomic analysis to detect changes in the expression levels of genes and proteins during russeting formation in russeted and non-russeted skin of 'Golden Delicious' apple. We set up three comparison groups representing the three developmental stages in the russeting formation process. With the formation of fruit russeting, there were 2856 differentially expressed genes and 942 differentially expressed proteins in the comparison groups as detected at the transcript level and protein level, respectively. A correlation analysis of the transcriptome and proteome data revealed related-genes involved in lignin biosynthesis are significant changes at different developmental stages during apple russeting formation. Some other transcription factors, such as MYBs, NACs and LIMs were also involved in apple russeting formation. In this study, one LIM transcription factor was preliminarily determined to be involved in lignin biosynthesis by combining to PAL-box element. Studying the identified genes and proteins will provide further insights into the molecular mechanisms controlling apple russeting formation.
Project description:DNA methylation is a key epigenetic modification in mammals, have essential and important roles in muscle development. Here, using cattle as a model, we investigate the systematic association between DNA methylation and muscle development. We sample longissimus thoracis tissues from a famous elite native breed of Chinese Qinchuan cattle living within comparable environments under fetal and adult stages. Using methylated DNA immunoprecipitation sequencing (MeDIP-Seq) , we generate and provide a genome-wide landscapes of DNA methylomes for muscle studies. The analysis shows global similarity and difference among fetal and adult stage and identifies the differentially methylated regions. The differentially methylated regions in promoters are highly associated with muscle development via expression repression of both known muscle-related genes and novel genes. This comprehensive map provides a solid basis for exploring epigenetic mechanisms of muscle growth and development. Examination of 2 different developmental stages of the Longissimus Muscle