• Title/Summary/Keyword: Oligonucleotide chip.

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Development of DNA Chip System for Differential Diagnosis of Porcine Enteric Pathogens

  • Kim, Tae-ju;Cho, Ho-seong;Kim, Yong-hwan;A.W.M. Effendy;Park, Nam-yong
    • Proceedings of the Korean Society of Veterinary Pathology Conference
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    • 2003.10a
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    • pp.32-32
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    • 2003
  • Intestinal infections are common in growing pigs and can be caused by multiple pathogens, environmental and management factors [1]. Among the most important viruses in swine enteritis are porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine enteric calicivirus (PECV), porcine group A rotavirus (PRV gp A) and bacteria are Escherichia coli and Salmonella spp. and protozoa is Isospora suis [1]. The DNA chip system can serve as a powerful tool that can be utilized for simultaneous detection of specific pathogenic bacteria strains and viruses [2,3]. The combination of PCR and DNA chip technology will provide a novel method for the detection of porcine enteric pathogens thus revolutionize the diagnosis and management of the disease. The aim of this study is to develop DNA chip system for the rapid and reliable detection of five major porcine enteric pathogens based on oligonucleotide DNA chip hybridization. (omitted)

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Effect of Carthami Tinctorii Fructus Herbal-acupuncture Solution(CTF-HAS) on Gene Expression in HepG2 carcinomar cells (Oligonucleotide chip를 이용한 홍화자약침액(紅花子藥鍼液)이 간암세포주(肝癌細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響))

  • Lee, Kyung-min;Lim, Seong-chul;Jung, Tae-young;Seo, Jung-chul;Han, Sang-won
    • Journal of Acupuncture Research
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    • v.22 no.3
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    • pp.215-225
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    • 2005
  • Objective : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells lines. Oligonucleotide microarray approach were employed to screen the differential expression genes. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. Cells were treated with various concentrations of CTF-HAS(0.1, 0.5, 1.5, 10, $20mg/m{\ell}$) for 24 h. Cytotoxicity was tested by MTT assay. To screen the differentially expressed genes in cancer cells, cells were treated with $1.5mg/m{\ell}$ of CTF-HAS. For oligonucleotide microarray assay, total RNA was used for gene expression analysis using oligonucleotide genechip (Human genome U133 Plus 2.0., Affimatrix Co.). ResuIts : It has no cytotoxic effects on HepG2 cells in all concentrations (0.1, 0.5, 1.5, 10, $20mg/m{\ell}$). More than twofold up-regulated genes were 19 genes. The number of more than twofold down-regulated genes was 13. Discussion : This study showed the screening of CTF-HAS mediated differentially regulated genes using combined approaches of oligonucleotide microarray. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.

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Nanoparticle-based Detection Technology for DNA Analysis

  • Park, Hyun-Gyu
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.8 no.4
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    • pp.221-226
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    • 2003
  • With the current rapid development of nanotechnology and synthesis technology for designed oligonucleotides or oligonucleotide-modified nanoparticle conjugates, the combined strategies have become one of the most valuable methods in detection technology for DNA analysis. Using the uniquely recognizable interactions of pre-designed DNA molecules in assembling nanoparticles, various novel approaches have been recently developed towards detecting specific DNA sequences. Here we describe the key fundamentals and issues of this promising strategies ranging from the initial findings of rationally designed DNA-based assembly of nanoparticles to the extended chip-based detection system. Some limitations of these new strategies and possible approaches will be also discussed for the practical application in the area of DNA microarray detection.

Differential Protein and Gene Expression after Adenovirus-Mediated p16 Gene Transfer in Human Non-Small Cell Lung Cancer Cells

  • Park, Mi-Sun;Kang , Ho-Il;Jee, Seung-Wan;Lim, Si-Nae;Pyo, Jae-Hee;Eom , Mi-Ok;Ryeom , Tai-Kyung;Kim, Ok-Hee
    • Proceedings of the PSK Conference
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    • 2002.10a
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    • pp.291.2-291.2
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    • 2002
  • For the safety evaluation of adenovirus-mediated gene therapy. we have investigated gene and protein expression after transduction of adenoviral vector (Ad5CMV-p16) which contains tumor suppressor gene. p161NK4$\alpha$ in human non-small cell lung cancer (A549) cells. We compared the differential gene expression level in the A549 cells treated with Ad5CMV (null type) and Ad5CMV-p16 virus. respectively. by using cDNA membrane chip and oligonucleotide chip. (omitted)

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Beet western yellows virus (BWYV): Aspect of Outbreak and Survey, and First Complete Genome Sequence of a Korea Isolate of BWYV

  • Park, Chung Youl;Kim, Jeong-Sun;Lee, Hong Kyu;Oh, Jonghee;Lim, Seungmo;Moon, Jae Sun;Lee, Su-Heon
    • Research in Plant Disease
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    • v.24 no.4
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    • pp.276-284
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    • 2018
  • In 2010, foliar symptoms were observed in the paprika leaves in Jinju city, Korea. Beet western yellows virus (BWYV) was identified in paprika by using the large-scale oligonucleotide chip assay. To investigate the occurrence of BWYV, a survey was performed on various crops, including paprika, from 2011 to 2014. Further, the presence of BWYV was consistently verified through literature survey from 2015 to 2017. BWYV infection has been identified in Solanaceae crops (bell pepper, hot pepper, and paprika), various weeds, and green peach aphids and it occurs on a nationwide scale. Cultivation using organic methods involved natural enemies and showed a high BWYV infection rate, which was more than that for conventional cultivation methods in greenhouse. The complete genome sequence of BWYV isolated from paprika was determined for the first time. The genome of the BWYV-Korea isolate consists of 5750 nucleotides and has six open reading frames. Sequence identity results showed maximum similarity between the BWYV-Korea isolate and the BWYV LS isolate (identity > 90%). This study is the first report of BWYV infecting paprika in Korea. The survey revealed that BWYV is naturalized in the domestic ecology of Korea.

Gene Expression Profile of Rat Hypothalamus Treated with Electroacupuncture at ST36 Acupoint (족삼리 전침자극에 의한 흰쥐 hypothalamus의 유전자 발현 profile 분석)

  • Rho Sam Woong;Lee Gi Seog;Choi Gi Soon;Na Young In;Hong Moo Chang;Shin Min Kyu;Min Byung il;Bae Hyun Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.4
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    • pp.1041-1054
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    • 2004
  • Electroacupuncture (EA) has been reported to increase pain threshold, and to enhance the NK cell activity by up-regulation of IFN-γ and endogenous β-endolphin. For the purpose of understanding the molecular mechanism of EA stimulation, we analyzed the gene expression profile of rat hypothalamus, treated on Zusanli (ST36) with EA, in comparison with control group by oligonucleotide chip microarray (Affymetrix GeneChip Rat Neurobiology U34 Array) and real-time RT-PCR. Sprague-Dawley (S-D) male rats were stimulated at the Zusanli (ST36) acupoint in restriction holder. Simultaneously the control group was given only holder stress without EA stimulation. In order to prove the appropriateness of EA treatment, we measured spleen NK cell activity with standard 51Cr release assay. NK cell activity of EA group was significantly increased comparing to control group. The microarray and PCR results show that EA treatment up-regulates expression of genes associated with 1) nerve growth such as NGF induced factor A and VGF, 2) signal transduction such as 5HT3 receptor subunit, AMPA receptor binding protein and Na-dependent neurotransmitter transporter, and 3) anti-oxidation such as superoxide dismutase and glutathione S-transferase. In addition, the activity of the anti-oxidative enzyme, SOD of hypothalamus, liver and RBC was enhanced compared to that of control. The list of differentially expressed genes may implicate further insight on the mechanism of acupuncture effects.

Gene Expression Profiling of Human Bronchial Epithelial (BEAS-2B) Cells Treated with Nitrofurantoin, a Pulmonary Toxicant

  • Kim, Youn-Jung;Song, Mee;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • v.3 no.4
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    • pp.222-230
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    • 2007
  • Some drugs may be limited in their clinical application due to their propensity towards their adverse effects. Toxicogenomic technology represents a useful approach for evaluating the toxic properties of new drug candidates early in the drug discovery process. Nitrofurantoin (NF) is clinical chemotherapeutic agent and antimicrobial and used to treatment of urinary tract infections. However, NF has been shown to result in pulmonary toxic effects. In this research, we revealed the changing expression gene profiles in BEAS-2B, human bronchial epithelial cell line, exposed to NF by using human oligonucleotide chip. Through the clustering analysis of gene expression profiles, we identified 136 up-regulated genes and 379 down-regulated genes changed by more than 2-fold by NF. This study identifies several interesting targets and functions in relation to NF-induced toxicity through a gene ontology analysis method including biological process, cellular components, molecular function and KEGG pathway.

Prevalence and Genotype Distribution of Human Papillomavirus in Cheonan, Korea

  • Kim, Jae Kyung;Jeon, Jae-Sik;Lee, Chong Heon;Kim, Jong Wan
    • Journal of Microbiology and Biotechnology
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    • v.24 no.8
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    • pp.1143-1147
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    • 2014
  • Human papillomavirus (HPV) infection is considered to play a critical role in the development of cervical carcinoma, which is the third most common cancer among Korean females. Here, we performed a baseline study of HPV infection and genotyping using an HPV DNA chip, which is a type of oligonucleotide microarray. A total of 6,855 cervical swab specimens from 5,494 women attending Dankook University Hospital Health Improvement Center in Cheonan, Korea between 2006 and 2012, originally collected for HPV infection screening, were genotyped for HPV. The extracted DNA from the cervical specimens was investigated by an HPV DNA chip designed to detect 41 different HPV types. HPV was identified as positive in 1,143 (16.7%) of the 6,855 samples. The most frequently detected HPV genotypes were HPV types 16, 53, 56, 58, 39, 52, 70, 84, 68, 62, 35, 54, 81, 18, and 30, in descending order of incidence. The proportions of single and multiple HPV infections in the HPV-positive specimens were 78.1% and 21.9%, respectively. The average age of HPV-positive patients was 39.9 years, with the positive rate of HPV being the highest in the 10-29 age group (20.6%). We report here on the prevalence and distribution of 41 different genotypes of HPV according to age among women in Cheonan, Korea. These data may be of use as baseline data for the assessment of public health-related issues and for the development of area-specific HPV vaccines.

Improving data reliability on oligonucleotide microarray

  • Yoon, Yeo-In;Lee, Young-Hak;Park, Jin-Hyun
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2004.11a
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    • pp.107-116
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    • 2004
  • The advent of microarray technologies gives an opportunity to moni tor the expression of ten thousands of genes, simultaneously. Such microarray data can be deteriorated by experimental errors and image artifacts, which generate non-negligible outliers that are estimated by 15% of typical microarray data. Thus, it is an important issue to detect and correct the se faulty probes prior to high-level data analysis such as classification or clustering. In this paper, we propose a systematic procedure for the detection of faulty probes and its proper correction in Genechip array based on multivariate statistical approaches. Principal component analysis (PCA), one of the most widely used multivariate statistical approaches, has been applied to construct a statistical correlation model with 20 pairs of probes for each gene. And, the faulty probes are identified by inspecting the squared prediction error (SPE) of each probe from the PCA model. Then, the outlying probes are reconstructed by the iterative optimization approach minimizing SPE. We used the public data presented from the gene chip project of human fibroblast cell. Through the application study, the proposed approach showed good performance for probe correction without removing faulty probes, which may be desirable in the viewpoint of the maximum use of data information.

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Effects of Inhibitors on the Function and Activity of Topoisomerase, and Gene Expression in HL-60 Human Leukemia Cells (HL-60 세포의 유전자 발현 및 topoisomerase의 기능 활성에 미치는 억제제의 영향)

  • Jeong, In-Cheol;Cho, Moo-Youn;Park, Jang-Su
    • Journal of Life Science
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    • v.18 no.1
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    • pp.75-83
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    • 2008
  • This studies were designed to elucidate whether inhibitors of topoisomerase regulate function and activity of topoisomerase, and gene expression in HL-60 human leukemia cells. HL-60 cells were treated with 10-hydroxycamptothecin or doxorubicin, total RNA was isolated, and expressed genes were investigated with human oligonucleotide microarray containing 10K gene, respectively. Expression profiles of the human leukemia HL-60 cells treated with 10-hydroxycamptothecin (10-CIT) or doxorubicin associated with signal transduction,. cell adhesion, cell cycle, cell growth, cell proliferation, cell differentiation, transcription and immune response, especially genes related with transcription and cell growth. In HL-60 cells treated with 10-CPT, the expression of topoisomerase III${\alpha}$, III${\beta}$ and I gene from oligo chip microarray analysis were increased over, but the expression of topoisomerase II${\alpha}$ and II${\beta}$ gene were decreased over. In contrast, the expression of topoisomerase II${\alpha}$ and II${\beta}$ gene were increased over in HL-60 cells treated with doxorubicin, whereas the expression of topoisomerase III${\alpha}$ and III${\beta}$ mRNA remained no significant change. These results suggest that these data may be useful for novel therapeutic markers.