• Title/Summary/Keyword: Gene Screening

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Cloning of the novel putative apoptosis-related gene of Spirometra erinacei (Order Pseudophyllidea)

  • Lee Soo-Ung;Huh Sun
    • Parasites, Hosts and Diseases
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    • v.44 no.3
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    • pp.233-237
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    • 2006
  • We postulated that apolysis was processed in accordance with apoptotic changes occurring in a cestode, Spirometra erinacei (Pseudophyllidea). We cloned the novel putative apoptosis-associated gene from S. erinacei via screening of a S. erinacei cDNA library with a ced-3 gene (activator of apoptosis) probe from Caenorhabditis elegans. We identified a 261-bp cDNA sequence, which encodes for an 86-amino acid protein. The cloned gene expression was observed in the neck and gravid proglottids via Northern blotting, using cloned cDNA inserts as probes, but the clone was not expressed in any of other tissues. We suggest that this gene may be involved in the apolysis of S. erinacei during normal tissue development and differentiation in cestode parasites.

Zoolan Gene Cloning of Zoogloea ramigera 115 (Zoogloea ramigera 115의 Zooglan Gene Cloning)

  • 이기영;전순배
    • KSBB Journal
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    • v.11 no.1
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    • pp.115-123
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    • 1996
  • Two kinds of mutants were isolated to clone a cluster of genes essential for zooglan biosynthesis. Zoogloea ramigera 115 strains produce capsular polysaccharide. To achieve conjugation in strain 115 and to facilitate recovery of product, a capsule non-forming strain was isolated via successive centrifugation and screening. The other kind of mutants devoid of or producing altered exopolysaccharides were obtained using classical transposon(Tn5) technique and screened for altered colony morphology and celluflour binding properties. Complementation of these mutants was achieved with Z. ramigera 115 slime gene library constructed in a broad host range cosmid vector and helper plasmid by triparental conjugation.

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Applying a modified AUC to gene ranking

  • Yu, Wenbao;Chang, Yuan-Chin Ivan;Park, Eunsik
    • Communications for Statistical Applications and Methods
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    • v.25 no.3
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    • pp.307-319
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    • 2018
  • High-throughput technologies enable the simultaneous evaluation of thousands of genes that could discriminate different subclasses of complex diseases. Ranking genes according to differential expression is an important screening step for follow-up analysis. Many statistical measures have been proposed for this purpose. A good ranked list should provide a stable rank (at least for top-ranked gene), and the top ranked genes should have a high power in differentiating different disease status. However, there is a lack of emphasis in the literature on ranking genes based on these two criteria simultaneously. To achieve the above two criteria simultaneously, we proposed to apply a previously reported metric, the modified area under the receiver operating characteristic cure, to gene ranking. The proposed ranking method is found to be promising in leading to a stable ranking list and good prediction performances of top ranked genes. The findings are illustrated through studies on both synthesized data and real microarray gene expression data. The proposed method is recommended for ranking genes or other biomarkers for high-dimensional omics studies.

Isolation of Small Prothoracicotropic Hormone-Like Gene in Drosophila mefanoguster (초파리에서 전홍선자극 호르몬 유사 유전자의 재조합)

  • Ki Wha Chung;Huu
    • The Korean Journal of Zoology
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    • v.37 no.1
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    • pp.12-18
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    • 1994
  • The prothoracicotropic hormone (PTTH) produced by the neurosecretory cells in insects is involved in molting and metamorphosis by activating the prothoracic frins) glands to secrete ecdysone (or related ecdvsteroidsl. In the present study, the small PTTH-like gene was isolated by screening of CDNA library using the bombvxin (corresponding to small PTTH in Bombvx moril gene probe in Drosophilo melonogaster. It showed 50-6096 sequence homology to bombyxin gene. The expression patterns of this gene showed developmental stage- and tissue-dependent manners. The mRNA was detected only in the late third instar larval-prepupa which is stases showing the highest hormonal activity to secrete ecdysteroids, and detected in the brain pan of the Isle third instar lanrae.

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BRCA1 Gene Mutation Screening for the Hereditary Breast and/or Ovarian Cancer Syndrome in Breast Cancer Cases: a First High Resolution DNA Melting Analysis in Indonesia

  • Mundhofir, Farmaditya EP;Wulandari, Catharina Endah;Prajoko, Yan Wisnu;Winarni, Tri Indah
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.3
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    • pp.1539-1546
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    • 2016
  • Specific patterns of the hereditary breast and ovarian cancer (HBOC) syndrome are related to mutations in the BRCA1 gene. One hundred unrelated breast cancer patients were interviewed to obtain clinical symptoms and signs, pedigree and familial history of HBOC syndrome related cancer. Subsequently, data were calculated using the Breast and Ovarian Analysis of Disease Incidence and Carrier Estimation Algorithm (BOADICEA) risk prediction model. Patients with high score of BOADICEA were offered genetic testing. Eleven patients with high score of BOADICEA, 2 patients with low score of BOADICEA, 2 patient's family members and 15 controls underwent BRCA1 genetic testing. Mutation screening using PCR-HRM was carried out in 22 exons (41 amplicons) of BRCA1 gene. Sanger sequencing was subjected in all samples with aberrant graph. This study identified 10 variants in the BRCA1 gene, consisting of 6 missense mutations (c.1480C>A, c.2612C>T, c.2566T>C, c.3113A>G, c.3548 A>G, c.4837 A>G), 3 synonymous mutations (c.2082 C>T, c.2311 T>C and c.4308T>C) and one intronic mutation (c.134+35 G>T). All variants tend to be polymorphisms and unclassified variants. However, no known pathogenic mutations were found.

Identification of Fruit-specific cDNAs in a Ripened Inodorus Melon Using Differential Screening and the Characterization of on Abscisic Acid Responsive Gene Homologue

  • Hong, Se-Ho;Kim, In-Jung;Chung, Won-Il
    • Journal of Plant Biotechnology
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    • v.4 no.1
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    • pp.7-15
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    • 2002
  • Eight cDNAs corresponding to fruit-specific genes were isolated from ripened melon through differential screening. Sequence comparison indicated that six of these cDNAs encoded proteins were previously characterized into aminocyclopropane-1-carboxylate (ACC) oxidase, abscisic acid, stress and ripening inducible (ASR) gene, RINC-H2 zinc finger protein, pyruvate decarboxylase, or polyubiquitin. RFS2 and RFS5 were the same clone encoding polyubiquitin. The other cDNAs showed no significant homology with known protein sequences. The ASR homologue (Asr1) gene was further characterized on the cDNA and genomic structure. The deduced amino acid sequence had similar characteristics to other plant ASR. The Asr1 genomic DNA consisted of 2 exons and 1 intron, which is similar to the structure of other plants ASR genes. The promoter region of the Asr1 gene contained several putative functional cis-elements such as an abscisic acid responsive element (ABRE), an ethylene responsive element (ERE), a C-box or DPBf-1 and 2, Myb binding sites, a low temperature responsive element (LTRE) and a metal responsive element (MRE). The findings imply that these elements may play important roles in the response to plant hormones and environmental stresses in the process of fruit development. The results of this study suggest that the expressions of fruit specific and ripening-related cDNAs are closely associated with the stress response.

A Novel Mutation in the MUT Gene in an Asymptomatic Newborn with Isolated Methylmalonic Acidemia (메틸말론산혈증 신생아의 MUT 유전자에서 발견된 새로운 돌연변이)

  • Kwak, Min Jung;Kim, Yoo-Mi
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.14 no.2
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    • pp.174-177
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    • 2014
  • Isolated methylmalonic acidemia (MMA) is a group of autosomal recessive inborn errors of metabolism caused by impaired activity of methylmalonyl-coenzyme A mutase (MCM). Mutations in the gene encoding MCM (MUT ) is the most common cause of isolated MMA. In this report, we identify an asymptomatic 15 days old female who had elevated C3-acylcarnitine (C3) in the newborn screening. Her serum homocysteine was normal and urine methylmalonic acid was increased that suggested isolated MMA. She was maintained on a low-protein diet and carnitine supplementation. At 3 months of age, she was still asymptomatic and had normal growth. We analyzed MUT gene mutations. Two heterozygote mutations in the MUT gene were identified including c.323G>A and c.1672+2T>C (IVS8(+2)T>C. Among these, c.1672+2T>C (IVS8(+2)T>C) have not been described previously.

Gene Expression in Zn-deficient U937 Cell Line : Using cDNA Microarray (아연결핍된 단핵구 U937 Cell Line에 있어서의 유전자 발현 탐색 : cDNA Microarray 기법 이용)

  • Beattie, John H.;Trayhurn, Paul
    • Journal of Nutrition and Health
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    • v.35 no.10
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    • pp.1053-1059
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    • 2002
  • In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.

Identification and Validation of Novel Biomarkers and Potential Targeted Drugs in Cholangiocarcinoma: Bioinformatics, Virtual Screening, and Biological Evaluation

  • Wang, Jiena;Zhu, Weiwei;Tu, Junxue;Zheng, Yihui
    • Journal of Microbiology and Biotechnology
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    • v.32 no.10
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    • pp.1262-1274
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    • 2022
  • Cholangiocarcinoma (CCA) is a complex and refractor type of cancer with global prevalence. Several barriers remain in CCA diagnosis, treatment, and prognosis. Therefore, exploring more biomarkers and therapeutic drugs for CCA management is necessary. CCA gene expression data was downloaded from the TCGA and GEO databases. KEGG enrichment, GO analysis, and protein-protein interaction network were used for hub gene identification. miRNA were predicted using Targetscan and validated according to several GEO databases. The relative RNA and miRNA expression levels and prognostic information were obtained from the GEPIA. The candidate drug was screened using pharmacophore-based virtual screening and validated by molecular modeling and through several in vitro studies. 301 differentially expressed genes (DEGs) were screened out. Complement and coagulation cascades-related genes (including AHSG, F2, TTR, and KNG1), and cell cycle-related genes (including CDK1, CCNB1, and KIAA0101) were considered as the hub genes in CCA progression. AHSG, F2, TTR, and KNG1 were found to be significantly decreased and the eight predicted miRNA targeting AHSG, F2, and TTR were increased in CCA patients. CDK1, CCNB1, and KIAA0101 were found to be significantly abundant in CCA patients. In addition, Molport-003-703-800, which is a compound that is derived from pharmacophores-based virtual screening, could directly bind to CDK1 and exhibited anti-tumor activity in cholangiocarcinoma cells. AHSG, F2, TTR, and KNG1 could be novel biomarkers for CCA. Molport-003-703-800 targets CDK1 and work as potential cell cycle inhibitors, thereby having potential for consideration for new chemotherapeutics for CCA.

Application of digital polymerase chain reaction technology for noninvasive prenatal test

  • Lee, Seung Yong;Hwang, Seung Yong
    • Journal of Genetic Medicine
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    • v.12 no.2
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    • pp.72-78
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    • 2015
  • Recently, noninvasive prenatal test (NIPT) has been adopted as a primary screening tool for fetal chromosomal aneuploidy. The principle of NIPT lies in isolating the fetal fraction of cell-free DNA in maternal plasma and analyzing it with bioinformatic tools to measure the amount of gene from the target chromosome, such as chromosomes 21, 18, and 13. NIPT will contribute to decreasing the need for unnecessary invasive procedures, including amniocentesis and chorionic villi sampling, for confirming fetal aneuploidy because of its higher positive predictive value than that of the conventional prenatal screening method. However, its greater cost than that of the current antenatal screening protocol may be an obstacle to the adoption of this innovative technique in clinical practice. Digital polymerase chain reaction (dPCR) is a novel approach for detecting and quantifying nucleic acid. dPCR provides real-time diagnostic advantages with higher sensitivity, accuracy, and absolute quantification than conventional quantitative PCR. Since the groundbreaking discovery that fetal cell-free nucleic acid exists in maternal plasma was reported, dPCR has been used for the quantification of fetal DNA and for screening for fetal aneuploidy. It has been suggested that dPCR will decrease the cost by targeting specific sequences in the target chromosome, and dPCR-based noninvasive testing will facilitate progress toward the implementation of a noninvasive approach for screening for trisomy 21, 18, and 13. In this review, we highlight the principle of dPCR and discuss its future implications in clinical practice.