Project description:The sheep (Ovis aries) plays a major socio-economic role in the world. Copy number variations (CNVs) are increasingly recognized as a key and potent source of genetic variation and phenotypic diversity, but little is known about the extent to which CNVs contribute to genetic variation in Chinese sheep breeds. Analyses of CNVs in the genomes of eight sheep breeds were performed using the sheep SNP50 BeadChip genotyping array. A total of 111 CNV regions (CNVRs) were obtained from 160 Chinese sheep breeds. These CNVRs covered 13.75 Mb of the sheep genome sequence. A total of 22 Go terms and 17 candidate genes were obtained from the functional analysis. Ten CNVRs were selected for validation, of which 7 CNVRs were further experimentally confirmed by quantitative PCR. Four candidate genes were selected to confirm the results of the functional analysis. These results provide a resource for furthering understanding of ruminant biology, and for further improving the genetic quality of sheep breeds.
Project description:We report the application of Solexa high-throuthput sequencing technology for miRNA differential expression profiles. Using an optimized data analysis workflow, we mapped about 30 million sequence reads per sample to Ovis aries genome. The results were validated by qRT-PCR. Our study represents the first detailed analysis of miRNA expression profiles in goat mammary gland tissues, with biologic replicates generated by RNA-seq technology.
Project description:Purpose: Aim of this study was to determine the miRNAs associated with average daily gain in Kangal Akkaraman lambs. Depending on the average daily gain, two groups were formed as low average daily gain (LDG) and high average daily gain (HDG). Methods: Peripheral blood mononuclear cells (pbmc) were separated from blood samples obtained from lambs and RNA was isolated with TRI Reagent (Sigma). Isolated total RNAs from LDG (n = 17) and HDG (n = 21) groups were pooled. miRNA libraries (LDG and HDG groups) were prepared with NEBNext® Multiplex Small RNA Library Prep Set for Illumina® (NEB, USA.) and sequenced with Illumina Novaseq. Readings were mapped with the Bowtie program using the Oar_v3.1 sheep reference genome. Differential expression analysis of the two groups was performed using the DESeq R package (1.8.3). Results: Using an optimized data analysis workflow, we mapped about 18 million sequence reads in HDG and 21 million sequence reads n LDG samples to the ovis aries genome (oar_v3.1). In total, 129 known miRNAs were identified, 134 novel miRNAs were predicted, and 40 differentially expressed miRNAs were found between the groups. Conclusions: Our study represents the first detailed analysis of Kangal type Akkaraman sheep PBMC miRNAs with RNA-seq technology. The obtained sequence reads would provide data for futher breeding studies related to average daily gain in Kangal Akkraman sheep.
Project description:Here, we analyzed and identified the miRNA expression profile of three different intestinal tissues (i.e., duodenum, cecum, and colon) of sheep (Ovis aries) using high-throughput sequencing and bioinformatic methods. In total, 128 known miRNAs were identified, 526 novel miRNAs were predicted, and 202 differentially expressed miRNAs were found between the different tissues. Additionally, 4,422 candidate target genes were predicted, and 185 non-redundant GO annotation terms were identified using enrichment analysis. A total of 529 target genes were found to participate in 37 KEGG biological pathways, and 270 of these genes were significantly enriched in the metabolism category.
Project description:Milk can mediate maternal-neonatal signal transmission by the bioactive component-extracellular vesicles (EVs), which select specific types of miRNA to encapsulate. The miRNA profiling of sheep milk EVs was characterized by sequencing and compared with that of cow milk. Sheep milk EVs contained various small RNAs, including tRNA, Cis-regulatory element, rRNA, snRNA, other Rfam RNA, and miRNA, which held about 36% of all the small RNAs. Totally 84 types of miRNAs were annotated with Ovis aries by miRBase (version 22.0) in sheep milk EVs, with 75 shared types of miRNAs in all samples. Fourteen sheep milk EV-miRNAs in the top 20, occupying 98% of the total expression, were immune-related.
Project description:Here we present a high-density in situ synthesized microarray for Ovis aries, named Aristaeus, designed by means of a pipeline of software instruments that, starting from non-annotated redundant EST sequences, selects oligonucleotides suitable for in situ generation on chip. The chip was tested by comparing the gene expression profiles of two sheep breeds with different phenotype, Sarda and Gentile di Puglia. We carried out microarray experiments on liver and udder tissues from lactating individuals and identified a relevant number of differentially expressed genes, all involved in metabolism pathways. The results are consistent with literature knowledge, while selected differential gene expressions have been confirmed by quantitative real-time polymerase chain reaction analyses. Tissue samples of liver were collected from 4 lactating individuals of two sheep (Ovis aries) breeds, Gentile di Puglia and Sarda. Biopsies of liver tissue were taken at second lactation stage (first record, stage 01: 6 days after lambing; second record, stage 02: 44 days after lambing) in both breeds. Tissues from liver were immersed in RNAlater (Sigma) immediately after biopsy and stored at -20°C. Samples were pooled by breed and then reverse labeled (cy5 and cy3), resulting in four raw data sets.
Project description:Milk and dairy products are an essential food and an economic resource in many countries. Milk component synthesis and secretion by the mammary gland involve expression of a large number of genes whose nutritional regulation remains poorly defined. We aim at understanding the genomic influence on milk quality and synthesis by comparing two sheep breeds, with different milking attitude, Sarda and Gentile di Puglia, using sheep-specific microarray technology. From sheep ESTs deposited at NCBI, we generated the first annotated microarray developed for sheep with a covering of most of the genome. Whole tissue samples of mammary gland were collected from 4 lactating individuals of two sheep (Ovis aries) breeds, Gentile di Puglia and Sarda. Biopsies of lactating mammary tissue were taken at two lactation stages (first record, stage 01: 6 days after lambing; second record, stage 02: 44 days after lambing) in both breeds. Tissues from mammary gland were immersed in RNAlater (Sigma) immediately after biopsy and stored at -20°C.
Project description:We report the application of Solexa high-throuthput sequencing technology for miRNA differential expression profiles. Using an optimized data analysis workflow, we mapped about 30 million sequence reads per sample to Ovis aries genome. The results were validated by qRT-PCR. Our study represents the first detailed analysis of miRNA expression profiles in goat mammary gland tissues, with biologic replicates generated by RNA-seq technology. miRNA expression profiles of goat mammary gland tissues between peak lactation and late lactation were generated by Illumina/Solexa sequencing