• Title/Summary/Keyword: Target enrichment

Search Result 84, Processing Time 0.023 seconds

A Simple and Efficient Subtractive Cloning Method

  • Min, Hyun-Jin;Park, Sang-Soo;Cho, Tae-Ju
    • BMB Reports
    • /
    • v.34 no.1
    • /
    • pp.59-65
    • /
    • 2001
  • In subtractive hybridization, target sequences in the tester are enriched by hybridizing with an excess amount of driver, followed by removing the tester hybridized with the driver. All of existing subtractive cloning methods are designed to remove the tester/driver hybrid. The removal of hybrid, however, is often unsatisfactory For various reasons. In this study we developed a subtractive enrichment protocol in which the tester/driver can be completely removed by selecting only the tester/tester after hybridization. In this protocol both the tester and driver DNAs are ligated with same linker DNAs and amplified by polymerase chain reaction (PCR). The tester DNA is then digested with two different enzymes and used in subsequent hybridization with an excess driver. After hybridization, the DNA is ligated with the adaptor that is only compatible with the tester/tester. Since only the tester/tester can have the new adaptor, no tester/driver can be amplified by PCR in this protocol. Unlike other methods, a 100% subtraction efficiency can be achieved even though the enzymatic treatments used in the enrichment procedure are incomplete. Furthermore, only the hybridized tester DNA can have the new adaptor and be amplified by PCR, resulting in 100% denaturation in effect. The efficacy of this novel method was verified with the model system in which a known amount of the target sequence is included.

  • PDF

MicroRNA Expression Profile Analysis Reveals Diagnostic Biomarker for Human Prostate Cancer

  • Liu, Dong-Fu;Wu, Ji-Tao;Wang, Jian-Ming;Liu, Qing-Zuo;Gao, Zhen-Li;Liu, Yun-Xiang
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.13 no.7
    • /
    • pp.3313-3317
    • /
    • 2012
  • Prostate cancer is a highly prevalent disease in older men of the western world. MicroRNAs (miRNAs) are small RNA molecules that regulate gene expression via posttranscriptional inhibition of protein synthesis. To identify the diagnostic potential of miRNAs in prostate cancer, we downloaded the miRNA expression profile of prostate cancer from the GEO database and analysed the differentially expressed miRNAs (DE-miRNAs) in prostate cancerous tissue compared to non-cancerous tissue. Then, the targets of these DE-miRNAs were extracted from the database and mapped to the STRING and KEGG databases for network construction and pathway enrichment analysis. We identified a total of 16 miRNAs that showed a significant differential expression in cancer samples. A total of 9 target genes corresponding to 3 DE-miRNAs were obtained. After network and pathway enrichment analysis, we finally demonstrated that miR-20 appears to play an important role in the regulation of prostate cancer onset. MiR-20 as single biomarker or in combination could be useful in the diagnosis of prostate cancer. We anticipate our study could provide the groundwork for further experiments.

Brain Plasticity and Stroke Rehabilitation (뇌가소성과 뇌졸중 재활)

  • Kim, Sik-Hyun
    • PNF and Movement
    • /
    • v.6 no.2
    • /
    • pp.39-50
    • /
    • 2008
  • Purpose : This article reviewed the advances in the understanding of the effect of motor rehabilitation and brain plasticity on functional recovery after CNS damage. Methods : This is literature study with Pubmed, Medline and Science journal. Results : The inability of CNS neurons to regenerate is largely associated with nonneuronal aspects of the CNS environment. Especially, this neuronal growth inhibition is mediated by myelin associated glycoprotein, olygodendrocyte-myelin glycoprotein, and NOGO. Enriched environment, motor learning, forced limb use have been utilized in scientific studies to promote functional reorganization and brain plasticity. Especially, enriched environment and motor enrichment may prime the brain to respond more adaptively to injury, in part by expressed neurotrophic factors. Conclusions : These reviews suggest that activity-induced neural plasticity occur in damaged brain areas in order to functional reorganization, where it could contribute to motor recovery, and represent a target for stroke rehabilitation.

  • PDF

Ultra-Specific Enrichment of GST-Tagged Protein by GSH-Modified Nanoparticles

  • Lee, Yeon-Ji;Park, Jong-Moon;Huh, Ji-Young;Kim, Min-Sik;Lee, Je-Sun;Palani, Arudra;Lee, Kwang-Yeol;Lee, Sang-Won
    • Bulletin of the Korean Chemical Society
    • /
    • v.31 no.6
    • /
    • pp.1568-1572
    • /
    • 2010
  • The selective isolation of specific proteins from complex protein mixtures by nanoparticles is reported. Glutathionemodified superparamagnetic nanoparticles were used to purify specific proteins fused with glutathione S-transferase via enzyme-substrate interactions. They demonstrated greatly improved selectivity and efficiency over micron sized capturing beads. The ultra-specific enrichment of target proteins was confirmed by both SDS-PAGE and LC/MS/MS experiments.

Inferring candidate regulatory networks in human breast cancer cells

  • Jung, Ju-Hyun;Lee, Do-Heon
    • Bioinformatics and Biosystems
    • /
    • v.2 no.1
    • /
    • pp.24-27
    • /
    • 2007
  • Human cell regulatory mechanism is one of suspicious problems among biologists. Here we tried to uncover the human breast cancer cell regulatory mechanism from gene expression data (Marc J. Van de vijver, et. al., 2002) using a module network algorithm which is suggested by Segal, et. al.(2003) Finally, we derived a module network which consists of 50 modules and 10 tree depths. Moreover, to validate this candidate network, we applied a GO enrichment test and known transcription factor-target relationships from Transfac(R) (V. Matys, et. al, 2006) and HPRD database (Peri, S. et al., 2003).

  • PDF

Target engagement of ginsenosides in mild cognitive impairment using mass spectrometry-based drug affinity responsive target stability

  • Zhu, Zhu;Li, Ruimei;Qin, Wei;Zhang, Hantao;Cheng, Yao;Chen, Feiyan;Chen, Cuihua;Chen, Lin;Zhao, Yunan
    • Journal of Ginseng Research
    • /
    • v.46 no.6
    • /
    • pp.750-758
    • /
    • 2022
  • Background: Mild cognitive impairment (MCI) is a transitional condition between normality and dementia. Ginseng is known to have effects on attenuating cognitive deficits in neurogenerative diseases. Ginsenosides are the main bioactive component of ginseng, and their protein targets have not been fully understood. Furthermore, no thorough analysis is reported in ginsenoside-related protein targets in MCI. Methods: The candidate protein targets of ginsenosides in brain tissues were identified by drug affinity responsive target stability (DARTS) coupled with label-free liquid chromatography-mass spectrometry (LC-MS) analysis. Network pharmacology approach was used to collect the therapeutic targets for MCI. Based on the above-mentioned overlapping targets, we built up a proteineprotein interaction (PPI) network in STRING database and conducted gene ontology (GO) enrichment analysis. Finally, we assessed the effects of ginseng total saponins (GTS) and different ginsenosides on mitochondrial function by measuring the activity of the mitochondrial respiratory chain complex and performing molecular docking. Results: We screened 2526 MCI-related protein targets by databases and 349 ginsenoside-related protein targets by DARTS. On the basis of these 81 overlapping genes, enrichment analysis showed the mitochondria played an important role in GTS-mediated MCI pharmacological process. Mitochondrial function analysis showed GTS, protopanaxatriol (PPT), and Rd increased the activities of complex I in a dose-dependent manner. Molecular docking also predicted the docking pockets between PPT or Rd and mitochondrial respiratory chain complex I. Conclusion: This study indicated that ginsenosides might alleviate MCI by targeting respiratory chain complex I and regulating mitochondrial function, supporting ginseng's therapeutic application in cognitive deficits.

Nucleic Acid Aptamers: New Methods for Selection, Stabilization, and Application in Biomedical Science

  • Kong, Hoon Young;Byun, Jonghoe
    • Biomolecules & Therapeutics
    • /
    • v.21 no.6
    • /
    • pp.423-434
    • /
    • 2013
  • The adoption of oligonucleotide aptamer is well on the rise, serving an ever increasing demand for versatility in biomedical field. Through the SELEX (Systematic Evolution of Ligands by EXponential enrichment), aptamer that can bind to specific target with high affinity and specificity can be obtained. Aptamers are single-stranded nucleic acid molecules that can fold into complex three-dimensional structures, forming binding pockets and clefts for the specific recognition and tight binding of any given molecular target. Recently, aptamers have attracted much attention because they not only have all of the advantages of antibodies, but also have unique merits such as thermal stability, ease of synthesis, reversibility, and little immunogenicity. The advent of novel technologies is revolutionizing aptamer applications. Aptamers can be easily modified by various chemical reactions to introduce functional groups and/or nucleotide extensions. They can also be conjugated to therapeutic molecules such as drugs, drug containing carriers, toxins, or photosensitizers. Here, we discuss new SELEX strategies and stabilization methods as well as applications in drug delivery and molecular imaging.

Systems-level mechanisms of action of Panax ginseng: a network pharmacological approach

  • Park, Sa-Yoon;Park, Ji-Hun;Kim, Hyo-Su;Lee, Choong-Yeol;Lee, Hae-Jeung;Kang, Ki Sung;Kim, Chang-Eop
    • Journal of Ginseng Research
    • /
    • v.42 no.1
    • /
    • pp.98-106
    • /
    • 2018
  • Panax ginseng has been used since ancient times based on the traditional Asian medicine theory and clinical experiences, and currently, is one of the most popular herbs in the world. To date, most of the studies concerning P. ginseng have focused on specific mechanisms of action of individual constituents. However, in spite of many studies on the molecular mechanisms of P. ginseng, it still remains unclear how multiple active ingredients of P. ginseng interact with multiple targets simultaneously, giving the multidimensional effects on various conditions and diseases. In order to decipher the systems-level mechanism of multiple ingredients of P. ginseng, a novel approach is needed beyond conventional reductive analysis. We aim to review the systems-level mechanism of P. ginseng by adopting novel analytical framework-network pharmacology. Here, we constructed a compound-target network of P. ginseng using experimentally validated and machine learning-based prediction results. The targets of the network were analyzed in terms of related biological process, pathways, and diseases. The majority of targets were found to be related with primary metabolic process, signal transduction, nitrogen compound metabolic process, blood circulation, immune system process, cell-cell signaling, biosynthetic process, and neurological system process. In pathway enrichment analysis of targets, mainly the terms related with neural activity showed significant enrichment and formed a cluster. Finally, relative degrees analysis for the target-disease association of P. ginseng revealed several categories of related diseases, including respiratory, psychiatric, and cardiovascular diseases.

Identifying Differentially Expressed Genes and Small Molecule Drugs for Prostate Cancer by a Bioinformatics Strategy

  • Li, Jian;Xu, Ya-Hong;Lu, Yi;Ma, Xiao-Ping;Chen, Ping;Luo, Shun-Wen;Jia, Zhi-Gang;Liu, Yang;Guo, Yu
    • Asian Pacific Journal of Cancer Prevention
    • /
    • v.14 no.9
    • /
    • pp.5281-5286
    • /
    • 2013
  • Purpose: Prostate cancer caused by the abnormal disorderly growth of prostatic acinar cells is the most prevalent cancer of men in western countries. We aimed to screen out differentially expressed genes (DEGs) and explore small molecule drugs for prostate cancer. Materials and Methods: The GSE3824 gene expression profile of prostate cancer was downloaded from Gene Expression Omnibus database which including 21 normal samples and 18 prostate cancer cells. The DEGs were identified by Limma package in R language and gene ontology and pathway enrichment analyses were performed. In addition, potential regulatory microRNAs and the target sites of the transcription factors were screened out based on the molecular signature database. In addition, the DEGs were mapped to the connectivity map database to identify potential small molecule drugs. Results: A total of 6,588 genes were filtered as DEGs between normal and prostate cancer samples. Examples such as ITGB6, ITGB3, ITGAV and ITGA2 may induce prostate cancer through actions on the focal adhesion pathway. Furthermore, the transcription factor, SP1, and its target genes ARHGAP26 and USF1 were identified. The most significant microRNA, MIR-506, was screened and found to regulate genes including ITGB1 and ITGB3. Additionally, small molecules MS-275, 8-azaguanine and pyrvinium were discovered to have the potential to repair the disordered metabolic pathways, abd furthermore to remedy prostate cancer. Conclusions: The results of our analysis bear on the mechanism of prostate cancer and allow screening for small molecular drugs for this cancer. The findings have the potential for future use in the clinic for treatment of prostate cancer.

On the Improvement of the Combustibility of Waste Plastics used in Blast Furnace

  • Ban, Bong-Chan;Choi, Jin-Shik;Kim, Dong-Su
    • Proceedings of the IEEK Conference
    • /
    • 2001.10a
    • /
    • pp.751-754
    • /
    • 2001
  • A possibility of using waste plastics as a source of secondary fuel in blast furnace has been of recent interest. The success of this process, however, will be critically dependent upon the optimization of operating systems. for instance, the supply of waste plastics must be reliable as well as economically attractive compared with conventional secondary fuels such as heavy oil, natural gas and pulverized coal. In this work, we put special importance on the improvement of the combustibility of waste plastics as a way to enhance energy efficiency in blast furnace. As experimental variables to approach this target, the effects of plastic particle size, blast temperature, and the level of oxygen enrichment were investigated using a custom-made blast model designed to simulate a real furnace. Lastly, the combustion efficiency of the mixture of waste plastics and pulverized coal was tested. The observations made from these experiments led us to the conclusion that with the increase of both blast temperature and the level of oxygen enrichment, and with the decrease of particle size, the combustibility of waste PE could be improved at a given distance from tuyere. Also it was found that the efficiency of coal combustion decreased with the addition of plastics; however, the combustion efficiency of mixture could be comparable at longer distance from tuyere.

  • PDF