• Title/Summary/Keyword: Target enrichment

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Development of the DGRS enriched in the high frequency range for APR1400 (고진등수 영역이 보강된 APR1400 설계지반응답스펙트럼의 개발)

  • 장영선;김태영;주광호;김종학
    • Proceedings of the Earthquake Engineering Society of Korea Conference
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    • 2001.09a
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    • pp.67-74
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    • 2001
  • This paper presents the Safe Shutdown Earthquake(SSE) input motion for the seismic design of the Advanced Power Reactor 1400(APR1400). The Design Ground Response Spectra(DGRS) far the SSE is based on the design spectrum specified in regulatory Guide(RG) 1.60 of U.S. Nuclear Regulatory Commission(US NRC), anchored to a Peak Ground Acceleration(PGA) of 0.3g and enriched in the high frequency range. This SSE seismic input motion is to be applied to the seismic analysis as the free-field seismic motion at the ground surface of both the rock and generic soil sites fur APRI1400. The enrichment for APR1400 seismic input motion is performed considering the current US NRC regulations, the seismic hazard studies performed by the Lawrence Livermore National Laboratory (LINL) and Electric Power Research Institute(EPRI) for the Central and Eastern United States nuclear power plant sites, and the seismic input motions used in the design certifications of the three existing U.S. advanced standard plants. It is represented by a set of DGRS and the accompanying Target Power Spectral Density(PSD) Function in both the horizontal and vertical directions.

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hnRNPK-regulated PTOV1-AS1 modulates heme oxygenase-1 expression via miR-1207-5p

  • Shin, Chang Hoon;Ryu, Seongho;Kim, Hyeon Ho
    • BMB Reports
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    • v.50 no.4
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    • pp.220-225
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    • 2017
  • Antisense transcripts were initially identified as transcriptional noise, but have since been reported to play an important role in the quality control of miRNA functions. In this report, we tested the hypothesis that heterogeneous nuclear ribonucleoprotein K (hnRNPK) regulates miRNA function via competitive endogenous RNAs, such as pseudogenes, long non-coding RNAs, and antisense transcripts. Based on analyses of RNA sequencing data, the knockdown of hnRNPK decreased the antisense PTOV1-AS1 transcript which harbors five binding sites for miR-1207-5p. We identified heme oxygenase-1 (HO-1) mRNA as a novel target of miR-1207-5p by western blotting and Ago2 immunoprecipitation. The knockdown of hnRNPK or PTOV1-AS1 suppressed HO-1 expression by increasing the enrichment of HO-1 mRNA in miR-1207-5p-mediated miRISC. Downregulation of HO-1 by a miR-1207-5p mimic or knockdown of hnRNPK and PTOV1-AS1 inhibited the proliferation and clonogenic ability of HeLa cells. Taken together, our results demonstrate that hnRNPK-regulated PTOV1-AS1 modulates HO-1 expression via miR-1207-5p.

Identification and Characterization of Calcineurin Targets in Cryptococcus neoformans

  • Park, Hee-Soo;Heitman, Joseph;Cardenas, Maria E.
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.17-17
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    • 2016
  • Calcineurin governs stress survival, sexual differentiation, and virulence of the human fungal pathogen Cryptococcus neoformans. Herein, we identified and characterized calcineurin substrates in C. neoformans by employing phosphoproteomic $TiO_2$ enrichment and quantitative mass spectrometry. The identified targets include the zinc finger transcription factor Crz1 and proteins whose functions are linked to P-bodies/stress granules (PBs/SGs) and mRNA translation and decay, such as Pbp1 and Puf4. We show that Crz1 is a bona fide calcineurin substrate, and localization and transcriptional activity of Crz1 are controlled by calcineurin. Several of the calcineurin targets localized to PBs/SGs, including Puf4 and Pbp1, and are required for survival at high temperature and for virulence. Genetic epistasis analysis revealed that Crz1 and the novel targets Lhp1, Puf4, and Pbp1 function in a branched calcineurin pathway that orchestrates stress survival and virulence. These findings propose that calcineurin controls thermal stress and virulence at the transcriptional level via Crz1 and post-transcriptionally by regulating target factors involved in mRNA metabolism.

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A Universal Analysis Pipeline for Hybrid Capture-Based Targeted Sequencing Data with Unique Molecular Indexes

  • Kim, Min-Jung;Kim, Si-Cho;Kim, Young-Joon
    • Genomics & Informatics
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    • v.16 no.4
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    • pp.29.1-29.5
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    • 2018
  • Hybrid capture-based targeted sequencing is being used increasingly for genomic variant profiling in tumor patients. Unique molecular index (UMI) technology has recently been developed and helps to increase the accuracy of variant calling by minimizing polymerase chain reaction biases and sequencing errors. However, UMI-adopted targeted sequencing data analysis is slightly different from the methods for other types of omics data, and its pipeline for variant calling is still being optimized in various study groups for their own purposes. Due to this provincial usage of tools, our group built an analysis pipeline for global application to many studies of targeted sequencing generated with different methods. First, we generated hybrid capture-based data using genomic DNA extracted from tumor tissues of colorectal cancer patients. Sequencing libraries were prepared and pooled together, and an 8-plexed capture library was processed to the enrichment step before 150-bp paired-end sequencing with Illumina HiSeq series. For the analysis, we evaluated several published tools. We focused mainly on the compatibility of the input and output of each tool. Finally, our laboratory built an analysis pipeline specialized for UMI-adopted data. Through this pipeline, we were able to estimate even on-target rates and filtered consensus reads for more accurate variant calling. These results suggest the potential of our analysis pipeline in the precise examination of the quality and efficiency of conducted experiments.

Dual-Toehold-Probe-Mediated Exonuclease-III-Assisted Signal Recycles Integrated with CHA for Detection of mecA Gene Using a Personal Glucose Meter in Skin and Soft Tissue Infection

  • Jiaguang Su;Wenjun Zheng
    • Journal of Microbiology and Biotechnology
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    • v.33 no.12
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    • pp.1692-1697
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    • 2023
  • Staphylococcus aureus integrated with mecA gene, which codes for penicillin-binding protein 2a, is resistant to all penicillins and other beta-lactam antibiotics, resulting in poor treatment expectations in skin and soft tissue infections. The development of a simple, sensitive and portable biosensor for mecA gene analysis in S. aureus is urgently needed. Herein, we propose a dual-toehold-probe (sensing probe)-mediated exonuclease-III (Exo-III)-assisted signal recycling for portable detection of the mecA gene in S. aureus. When the target mecA gene is present, it hybridizes with the sensing probe, initiating Exo III-assisted dual signal recycles, which in turn release numerous "3" sequences. The released "3" sequences initiate catalytic hairpin amplification, resulting in the fixation of a sucrase-labeled H2 probe on the surface of magnetic beads (MBs). After magnet-based enrichment of an MB-H1-H2-sucrase complex and removal of a liquid supernatant containing free sucrase, the complex is then used to catalyze sucrose to glucose, which can be quantitatively detected by a personal glucose meter. With a limit of detection of 4.36 fM for mecA gene, the developed strategy exhibits high sensitivity. In addition, good selectivity and anti-interference capability were also attained with this method, making it promising for antibiotic tolerance analysis at the point-of-care.

Serum exosomal miR-192 serves as a potential detective biomarker for early pregnancy screening in sows

  • Ruonan Gao;Qingchun Li;Meiyu Qiu;Su Xie;Xiaomei Sun;Tao Huang
    • Animal Bioscience
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    • v.36 no.9
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    • pp.1336-1349
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    • 2023
  • Objective: The study was conducted to screen differentially expressed miRNAs in sows at early pregnancy by high-throughput sequencing and explore its mechanism of action on embryo implantation. Methods: The blood serum of pregnant and non-pregnant Landrace×Yorkshire sows were collected 14 days after artificial insemination, and exosomal miRNAs were purified for high throughput miRNA sequencing. The expression patterns of 10 differentially expressed (DE) miRNAs were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The qRT-PCR quantified the abundance of serum exosomal miR-192 in pregnant and control sows, and the diagnostic power was assessed by receiver operating characteristic (ROC) analysis. The target genes of DE miRNAs were predicted with bioinformatics software, and the functional and pathway enrichment analysis was performed on gene ontology and the Kyoto encyclopedia of genes and genomes terms. Furthermore, a luciferase reporter system was used to identify the target relation between miR-192 and integrin alpha 4 (ITGA4), a gene influencing embryo implantation in pigs. Finally, the expression levels of miRNAs and the target gene ITGA4 were analyzed by qRT-PCR, and western blot, with the proliferation of BeWo cells detected by cell counting kit-8 (CCK-8). Results: A total of 221 known miRNAs were detected in the libraries of the pregnant and non-pregnant sows, of which 55 were up-regulated and 67 were down-regulated in the pregnant individuals compared with the non-pregnant controls. From these, the expression patterns of 10 DE miRNAs were validated. The qRT-PCR analysis further confirmed a significantly higher expression of miR-192 in the serum exosomes extracted from pregnant sows, when compared to controls. The ROC analysis revealed that miR-192 provided excellent diagnostic accuracy for pregnancy (area under the ROC curve [AUC]=0.843; p>0.001). The dual-luciferase reporter assay indicated that miR-192 directly targeted ITGA4. The protein expression of ITGA4 was reduced in cells that overexpressed miR-192. Overexpression of miR-192 resulted in the decreased proliferation of BeWo cells and regulated the expression of cell cycle-related genes. Conclusion: Serum exosomal miR-192 could serve as a potential biomarker for early pregnancy in pigs. miR-192 targeted ITGA4 gene directly, and miR-192 can regulate cellular proliferation.

Soil Contamination of Heavy Metals in National Industrial Complexes, Korea (국내 주요 국가산업단지에서 중금속에 의한 토양오염)

  • Jeong, Tae-Uk;Cho, Eun-Jeong;Jeong, Jae-Eun;Ji, Hwa-Seong;Lee, Kyeong-Sim;Yoo, Pyung-Jong;Kim, Gi-Gon;Choi, Ji-Yeon;Park, Jong-Hwan;Kim, Seong-Heon;Heo, Jong-Soo;Seo, Dong-Cheol
    • Korean Journal of Environmental Agriculture
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    • v.34 no.2
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    • pp.69-76
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    • 2015
  • BACKGROUND: Contamination of soils by heavy metals is the serious environmental problem. In particular, industrial processing is one of the main sources of heavy metal contamination. The objective of this study was to investigate the distribution characteristics of heavy metals in soils collected from industrial complex. METHODS AND RESULTS: In this study, the soil contamination and enrichment factor (EF) of heavy metals were investigated in three national industrial complexes such as Yeosu, Ulsan and Sihwa Banwal industrial complexes. The target heavy metals includes Cd, Cu, As, Hg, Pb, Cr, Zn, and Ni. The results showed that the contents of Cd, Hg, Pb, Zn and Ni in Yeosu and the contents of Cu, As and Cr in Sihwa Banwal were higher than in any other industrial complex. The results of principal component analysis(PCA) in Yeosu, Ulsan and Sihwa Banwal complex could be explained up to approximately 81.4, 69.1 and 70.9% by two factor, respectively. Enrichment factors of Cd, Pb and Zn in all the investigated industrial complexes were above 1.0 that was the value judged to be a high contamination. And EF of Cr was above 1.0 in Sihwa Banwal complex. EF of Zn in all sites was generally high from the other heavy metals. CONCLUSION: Therefore, soils maybe significantly affected by heavy metals (especially, Cd, Pb and Zn) present in the emissions from industrial complexes.

Preparation of L-cysteine Salicylaldehyde Schiff-base Modified Macroporous Polystyrene Resin and Its Application to Determination of Trace Cadmium and Lead in Environmental Water Samples

  • Xie, Fazhi;Zhang, Fengjun;Xuan, Han;Ge, Yejun;Wang, Yin;Li, Guolian;Zhu, Lei;Oh, Won-Chun
    • Journal of the Korean Ceramic Society
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    • v.51 no.5
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    • pp.472-476
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    • 2014
  • In this work, a new method that utilizes L-cysteine salicylaldehyde Schiff-base modified macroporous polystyrene resin (PS-CSC) as an effective sorbent has been developed for preconcentration of trace cadmium and lead in environmental water samples. The effect of pH, the contact time, the elution conditions, the flow rate, the initial concentration of target metal ions, and the effects of interfering ions on the preconcentration of the analytes were investigated. The maximum adsorption capacity of PS-CSC under optimum conditions for cadmium and lead were found to be 6.03 - 18.17 mg/g and 12.58 - 36.13 mg/g when the initial concentration of metal ions between 5.0 - 90 mg/L. The limits of detection for cadmium and lead were 2.46 ng/L and $0.52{\mu}g/L$, with a preconcentration factor of 200. The developed method has been validated by analyzing certified reference material and successfully applied for the enrichment and determination of trace cadmium and lead from environmental water samples.

Electrochemical Detection of $17{\beta}-estradiol$ by using DNA Aptamer Immobilized Nanowell Gold Electrodes

  • Kim, Yeon-Seok;Jung, Ho-Sup;Lee, Hea-Yeon;Kawai, Tomoji;Gu, Man-Bock
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.88-92
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    • 2005
  • Aptamer is the single-stranded oligonucleotide which binds to various target molecules such as proteins, peptides, lipids and small organic molecules with high affinity and specificity. DNA aptamers specific for the $17{\beta}-estradiol$ were selected by SELEX (Systematic Evolution of Ligands by EXponential enrichment) process from a random DNA library. These DNA aptamers have a high affinity to $17{\beta}-estradiol$ as an endocrine disrupting chemical. Nanowell and $200{\mu}m$ gold electrode were used as substrate for DNA aptamer immobilization and electrochemical analysis. Especially, nanowell gold electrode was fabricated by e-beam lithography. The size of single nanowell is 130nm and 40,000 nanowells were deposited on one gold electrode. The immobilization method was based on the interaction between the biotinylated aptamer and streptavidin deposited on gold electrode previously. Immobilization procedure was optimized by surface plasma resonance (SPR) and electrochemical analysis. After the immobilization of DNA aptamer on streptavidin modified gold electrode, $17{\beta}-estradiol$ solution was treated on aptamer immobilized gold electrode. The current of gold electrode was decreased by the binding of $17{\beta}-estradiol$ to DNA aptamer immobilized on gold electrode. However, in negative control experiments of 1-aminoanthraquinone and 2-methoxynaphthalene, the current was rarely decreased. And more sensitive data was obtained from nanowell gold electrode comparing with $200{\mu}m$ gold electrode.

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Comparative Genomics Reveals the Core and Accessory Genomes of Streptomyces Species

  • Kim, Ji-Nu;Kim, Yeonbum;Jeong, Yujin;Roe, Jung-Hye;Kim, Byung-Gee;Cho, Byung-Kwan
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1599-1605
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    • 2015
  • The development of rapid and efficient genome sequencing methods has enabled us to study the evolutionary background of bacterial genetic information. Here, we present comparative genomic analysis of 17 Streptomyces species, for which the genome has been completely sequenced, using the pan-genome approach. The analysis revealed that 34,592 ortholog clusters constituted the pan-genome of these Streptomyces species, including 2,018 in the core genome, 11,743 in the dispensable genome, and 20,831 in the unique genome. The core genome was converged to a smaller number of genes than reported previously, with 3,096 gene families. Functional enrichment analysis showed that genes involved in transcription were most abundant in the Streptomyces pan-genome. Finally, we investigated core genes for the sigma factors, mycothiol biosynthesis pathway, and secondary metabolism pathways; our data showed that many genes involved in stress response and morphological differentiation were commonly expressed in Streptomyces species. Elucidation of the core genome offers a basis for understanding the functional evolution of Streptomyces species and provides insights into target selection for the construction of industrial strains.