Prion diseases are a class of transmissible fatal disorders. In order to identify alterations associated with the pathogenesis of prion diseases, several studies have been conducted involving differential gene expression analysis using cDNA libraries, mRNA differential displays, and gene microarrays. These genomic approaches may be useful for identifying genes that are differentially expressed in prion diseases and that may participate directly or indirectly in the pathogenesis of the disease. In this study, we compared the gene expression profiles of normal and CWD-infected TgElk mice using the GeneFishing differentially expressed gene (DEG) screening system and real-time PCR analysis. DEGs were screened using the ACP-based PCR method with GeneFishing synthesis. In order to validate candidate genes, we used quantitative PCR (qPCR), and eleven DEGs were identified. Five of these eleven DEGs were upregulated and two were downregulated in the CWD mice. The DEGs newly identified in this study may be useful for diagnosing and studying the pathogenesis of prion diseases.
Arsenic (As) is a toxic element that easily taken up by plants root. Several toxic forms of As disrupt plant metabolism by a series of cellular alterations. In this study, we applied annealing control primer (ACP)-based reverse transcriptase PCR (polymerase chain reaction) technique to identify differentially expressed genes (DEGs) in alfalfa roots in response to As stress. Two-week-old alfalfa seedlings were exposed to As treatment for 6 hours. DEGs were screened from As treated samples using the ACP-based technique. A total of six DEGs including heat shock protein, HSP 23, plastocyanin-like domain protein162, thioredoxin H-type 1 protein, protein MKS1, and NAD(P)H dehydrogenase B2 were identified in alfalfa roots under As stress. These genes have putative functions in abiotic stress homeostasis, antioxidant activity, and plant defense. These identified genes would be useful to increase As tolerance in alfalfa plants.
Muscle satellite cell (SC) is responsible for postnatal muscle growth, repair, and regeneration. Satellite cell is an im-portant source of multi-potent stem cell process and differentiation into adipogenic, myogenic, and osteoblastogenic. The objective of this study was to identify alter of transcriptome during differentiation in porcine satellite cell and to elevated transcriptome at different stages of postnatal development to gain insight into the differences in differ-entiated PSC. We used RNA-seq technique to investigate the transcriptomes during differentiation in pig muscle. Sequence reads were obtained from Illumina HiSeq2000. Differentially expressed genes (DEG) were detected by EdgeR. Gene ontology (GO) terms are powerful tool for unification among representation genes or products. In study of GO biological terms, functional annotation clustering involved in cell cycle, apoptosis, extracellular matrix, phosphoryla- tion, proteolysis, and cell signaling in differences stage. Taken together, these results would be contributed to a better understanding of muscle biology and processes underlying differentiation. Our results suggest that the source of DEGs could be better understanding of the mechanism of muscle differentiation and transdifferentiation.
In the gingival tissues of patients with periodontitis, inflammatory responses are mediated by a wide variety of genes. In this study, we screened for differentially expressed genes (DEGs) in periodontitis compared with normal tissue using an annealing control primer (ACP) system. By ACP RT-PCR analysis, we obtained about 160 amplicons, 8 of which were found to be differentially expressed. DEGs in patients with periodontitis were thus successfully and reliably identified by the ACP-based RT PCR technique. The DEGs identified in the screen may also enhance our understanding of the pathogenesis of periodontitis.
Placenta is the main nutrition source for the fetus during pregnancy. Thus, it has a pivotal function in the pregnant process. Many functions of the placenta have been elucidated. An abnormal placenta is associated with a high rate of pregnancy failure in somatic cloned bovine. Differentially expressed genes (DEGs) were examined in a comparison between normal and cloned bovine placenta using annealing control primer (ACP)-based GeneFishing PCR. Using 120 ACPs, nearly 80 genes were identified and the fragments of 42 DEGs were sequenced. 38 of these genes were known genes and four were unknown. To determine the DEGs result, six target clones expressing on one-side of a normal and a clone placenta were selected. Through an analysis of the target genes using the real-time PCR, the expressing pattern was found to be somewhat different from the DEGs. Additionally, several genes appeared with the same expression pattern. Taken together, this suggests that the target genes would be essential for research into what influences the placental formative mechanisms during fetal development.
Silencing of Dicer1 by siRNA did not inhibit development up to the blastocyst stage, but decreased expression of selected transcription factors, including Oct‐4, Sox2 and Nanog, suggesting that Dicer1 gene expression is associated with differentiation processes at the blastocyst stage (Cui et al., 2007). In order to get insights into genes which may be linked with microRNA system, we compared gene expression profiles in Gapdh and Dicer1 siRNA‐microinjected blastocysts using the Applied Biosystem microarray technology. Our data showed that 397 and 737 out of 16354 genes were up‐ and down‐regulated, respectively, following siRNA microinjection (p<0.05), including 24 up‐ and 28 downregulated transcription factors. Identification of genes that are preferentially expressed at particular Dicer1 knock down embryos provides insights into the complex gene regulatory networks that drive differentiation processes in embryos at blastocyst stage.
Aging causes thymus involution, and genes in thymus play an important role in the development of the immune system. In this study, we compared genes expressed in thymus of neonatal and peripubertal rats using annealing control primers (ACPs)-based GeneFishing polymerase chain reaction (PCR) and semiquantitative reverse transcription (RT)-PCR. We identified 10 differentially expressed genes (DEGs) with 20 ACPs. Of 10 DEGs, bystin-like, collagen type V alpha 1 (COL5A1), and T-cell receptor beta-chain segment 2 (TCRB2) that are related to immune-function were detected in rat thymus. Bystin-like and TCRB2 were up-regulated, while COL5A1 was down-regulated in peripubertal thymus. Semiquantitative RT-PCR confirmed postnatal changes in expression of bystin-like, COL5A1, and TCRB2. These results suggest that bystin-like, COL5A1, and TCRB2 could regulate immune function controlled in thymus as age increases.
Space has many distinguishable characteristics from earth such as strong cosmic radiation, microgravity, supervaccum and weak magnetic field. For this reason, space environments can be used an efficient mutagen for plant breeding nowadays. To identify the affected genes by condition in space with outer space, Brachypodium seeds were placed in the Russia Segment (RS) Biorisk module of International Space Station (ISS). Brachypodium distachyon is a model system for temperature grass, because they represent the characteristics for annual winter grass. Seeds and organs of plants carried by satellite or spacecraft to space can be genetically mutated by exposing space environment. We performed a duplicated RNA sequencing to profile the differentially expressed genes. As a results, about 700 genes were upregulated and 250 genes were downregulated by cosmic environments, respectively. In the molecular function category, protein kinase and transcription activity related genes were upregulated. Among the many transcription factors (TFs), stress related TFs such as ERF, NAC and WRKY were differentially expressed in space exposed samples. In the future, their expression will be identified by using qRT_PCR.
Exposure to ionizing radiation is regarded as a kind of abiotic stresses that can change the expression of genes in living organisms. This study aimed on investigating the variations in gene expressions induced by two different types of irradiations with different doses, which were low linear energy transfer (LET) gamma rays (100, 200, and 400 Gy) and high LET ion-beams (20, 40, and 80 Gy) on rice. RNA sequencing was carried out using the Illumina HiSeq-2500 platform. The average amount of reads were 4.8 Gb per individual, and 5 to 8% of the reads were removed after quality control. More than 90% of the RNA-seq reads were mapped to the rice reference genome sequence (IRGSP-1.0). A total of 247 differentially expressed genes (DEGs) were identified by comparison of the gene expression levels between the wildtype and the irradiated individuals. The 247 DEGs were divided into five modules and 27 intra-modular hub genes were found using the weighted correlation network analysis (WGCNA) method. The MEturquiose module had the most number of genes with 75 related to carbohydrate and small molecule metabolic processes. The co-expression network reconstructed using ARACNE (algorithm for reconstruction of accurate cellular networks) showed specific up- or down-regulation of the genes in each module according to the types and doses of radiation. This study will contribute to understanding the gene expression responses to ionizing irradiation.
적색 과육 '홍양' 품종에서 차등발현하는 유전자를 찾기 위하여 mirror orientation selection (MOS)과 결합된 suppression subtractive hybridization (SSH) 실험을 수행하였다. 그 결과, 288개의 cDNA clone을 확보하였으며, colony PCR을 통해 192개의 positive clone을 선발하였고, 이들을 sequencing하였다. NCBI/Genbank 데이터베이스의 BLAST 검색를