• Title/Summary/Keyword: Complementary DNA

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Detection of DNA Adduct Formed by Mitomycin C by $^{32}P$-Postlabelling ($^{32}P$-Postlabelling 방법을 이용한 미토마이신 C에 의하여 형성된 DNA adduct의 검출)

  • Jeong, Hye-Yun;Kim, Jae-Hyeon;Park, Chang-Won;Lee, Dong-Gwon
    • YAKHAK HOEJI
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    • v.40 no.4
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    • pp.442-448
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    • 1996
  • Mitomycin C(MMC) has been used as an anticancer drug and behaves as an alkylating agent forming covalent cross-link between complementary strands of double strand DNA. The purpose of this research was to determine number of DNA adducts, formed in vivo by Mitomycin C, in mouse organs. DNAs from liver, lung, brain and pancreas were isolated and used for $^{32}P$-postlabelling. The labeled nucleotides were separated by 2D-TLC and subjected to autoradiography. Numbers of MMC-DNA adducts were 9,9,5,4 in liver, pancreas, lung and brain, respectively.

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Complementary DNA Cloning and Nucleotide Sequence Analysis of Coat Protein Gene from TMV Tomato Strain (토마토에서 분리된 담배 모자이크 바이러스 외피단백질 유전자의 cDNA 클로닝 및 염기서열 분석)

  • 이청호;이영기;강신웅;박은경
    • Journal of the Korean Society of Tobacco Science
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    • v.18 no.2
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    • pp.101-106
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    • 1996
  • Tobacco mosaic virus (TMV) tomato strain was isolated from tomato "Seo-Kwang" in Korea. The virion was purified by density gradient centrifugation, and total viral RNA was isolated from the purified particles. Coat protein (CP) cDNA of the virus was synthesized by RT-PCR, and the purified cDNA fragment was subcloned to pBluescript II SK-. The analysis of nucleotide sequence showed that this cDNA was 693 nucleotides long from the insert of clone p1571 and p1572 which contain complete codons of the viral coat protein gene (474 nucleotides) and 3' untranslated region. The nucleotides of coat protein encoding cDNA of the strain were 6 nucleotides less than that of TMV common strain isolated from tobacco plant in Korea. The CP gene showed 70% maximum homology with that of the common strain in the nucleotide level and 86% maximum homology in amino acid level.cid level.

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Development of DNA Sensor Using Magnetic Iron Oxide Nanoparticle (자성 산화철(iron oxide) 나노입자를 이용한 DNA 센서 개발)

  • Nam, Ki-Chang;Song, Kwang-Soup
    • Journal of the Institute of Electronics Engineers of Korea SC
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    • v.48 no.6
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    • pp.51-56
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    • 2011
  • The surface of magnetic iron oxide nanoparticles (${\gamma}-Fe_2O_3$) is functionalized ($-NH_2$, -COOH) with bifunctional organic molecules and evaluated using FT-IR (Fourier transform infrared spectroscopy). We immobilize 21-base pair probe DNA and hybridize fluorescence-labeled (Cy5) target DNA onto the functionalized iron oxide nanoparticles. The fluorescence images obtained from a confocal microscopy show that the functionalized iron oxide nanoparticles should detect the hybridization of complementary and noncomplementary DNA.

Graphene Based Electrochemical DNA Biosensor for Detection of False Smut of Rice (Ustilaginoidea virens)

  • Rana, Kritika;Mittal, Jagjiwan;Narang, Jagriti;Mishra, Annu;Pudake, Ramesh Namdeo
    • The Plant Pathology Journal
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    • v.37 no.3
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    • pp.291-298
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    • 2021
  • False smut caused by Ustilaginoidea virens is an important rice fungal disease that significantly decreases its production. In the recent past, conventional methods have been developed for its detection that is time-consuming and need high-cost equipments. The research and development in nanotechnology have made it possible to assemble efficient recognition interfaces in biosensors. In this study, we present a simple, sensitive, and selective oxidized graphene-based geno-biosensor for the detection of rice false smut. The biosensor has been developed using a probe DNA as a biological recognition element on paper electrodes, and oxidized graphene to enhance the limit of detection and sensitivity of the sensor. Probe single-stranded DNA (ssDNA) and target ssDNA hybridization on the interface surface has been quantitatively measured with the electrochemical analysis tools namely, cyclic voltammetry, and linear sweep voltammetry. To confirm the selectivity of the device, probe hybridization with non-complementary ssDNA target has been studied. In our study, the developed sensor was able to detect up to 10 fM of target ssDNA. The paper electrodes were employed to produce an effective and cost-effective platform for the immobilization of the DNA and can be extended to design low-cost biosensors for the detection of the other plant pathogens.

Inhibition Effects of Persicaria amphibia (L.) Delarbre on Oxidative DNA Damage via ATM/Chk2/p53 pathway

  • So-Yeon Han;Hye-Jeong Park;Jeong-Yong Park;Seo-Hyun Yun;Mi-Ji Noh;Soo-Yeon Kim;Tae-Won Jang;Jae-Ho Park
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.52-52
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    • 2021
  • Persicaria amphibia as an England native plant, is a rhizomatous perennial, one of the rather amphibious plants. Its aquatic form contains water-soluble sugars, starch, and protein. P. amphibia have up to 18% tannins in stems and rhizomes. Previous studies have confirmed the anti-inflammatory activity of live bacteria roots, but no studies on bioactivity are known. DNA damage responses (DDRs) pathways are considered a crucial factor affecting the alleviation of cellular damage. The ataxia-telangiectasia mutated and Rad3 related (ATM) and checkpoint kinase 2 (Chk2) pathways are the main pathways of DNA damage response. Also, p53 is a key integrator of cellular response to oxidative DNA damage, contributing repair, or leading transcription including apoptosis. In the present study, we conducted an investigation into the inhibitory effects of P. amphibia on oxidative DNA damage for confirming potential to complementary medicine and therapies. In conclusion, P. amphibia can provide protective effects against double-stranded DNA break (DSB) caused by oxidative DNA damage.

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ChIP-seq Library Preparation and NGS Data Analysis Using the Galaxy Platform (ChIP-seq 라이브러리 제작 및 Galaxy 플랫폼을 이용한 NGS 데이터 분석)

  • Kang, Yujin;Kang, Jin;Kim, Yea Woon;Kim, AeRi
    • Journal of Life Science
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    • v.31 no.4
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    • pp.410-417
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    • 2021
  • Next-generation sequencing (NGS) is a high-throughput technique for sequencing large numbers of DNA fragments that are prepared from a genome. This sequencing technique has been used to elucidate whole genome sequences of living organisms and to analyze complementary DNA (cDNA) or chromatin immunoprecipitated DNA (ChIPed DNA) at the genome level. After NGS, the use of proper tools is important for processing and analyzing data with reasonable parameters. However, handling large-scale sequencing data and programing for data analysis can be difficult. The Galaxy platform, a public web service system, provides many different tools for NGS data analysis, and it allows researchers to analyze their data on a web browser with no deep knowledge about bioinformatics and/or programing. In this study, we explain the procedure for preparing chromatin immunoprecipitation-sequencing (ChIP-seq) libraries and steps for analyzing ChIP-seq data using the Galaxy platform. The data analysis steps include the NGS data upload to Galaxy, quality check of the NGS data, premapping processes, read mapping, the post-mapping process, peak-calling and visualization by window view, heatmaps, average profile, and correlation analysis. Analysis of our histone H3K4me1 ChIP-seq data in K562 cells shows that it correlates with public data. Thus, NGS data analysis using the Galaxy platform can provide an easy approach to bioinformatics.

Bayesian Variable Selection in the Proportional Hazard Model with Application to DNA Microarray Data

  • Lee, Kyeon-Eun;Mallick, Bani K.
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2005.09a
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    • pp.357-360
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    • 2005
  • In this paper we consider the well-known semiparametric proportional hazards (PH) models for survival analysis. These models are usually used with few covariates and many observations (subjects). But, for a typical setting of gene expression data from DNA microarray, we need to consider the case where the number of covariates p exceeds the number of samples n. For a given vector of response values which are times to event (death or censored times) and p gene expressions (covariates), we address the issue of how to reduce the dimension by selecting the significant genes. This approach enable us to estimate the survival curve when n < < p. In our approach, rather than fixing the number of selected genes, we will assign a prior distribution to this number. The approach creates additional flexibility by allowing the imposition of constraints, such as bounding the dimension via a prior, which in effect works as a penalty. To implement our methodology, we use a Markov Chain Monte Carlo (MCMC) method. We demonstrate the use of the methodology to diffuse large B-cell lymphoma (DLBCL) complementary DNA(cDNA) data.

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Complementary DNA Cloning and nucleotide Sequence Analysis of Coat Protein Gene from TMV Pepper Strain (고추에서 분리된 담배 모자이크 바이러스 외피단백질 유전자의 cDNA 클로닝 및 염기서열 분석)

  • 이영기;이청호;강신웅;박은경
    • Korean Journal Plant Pathology
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    • v.12 no.2
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    • pp.182-186
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    • 1996
  • 국내에서 재배되고 있는 고추(Capsicum annuum L.)로부터 분리된 TMV pepper 계통을 density gradient centrifugation을 이용하여 순화하였다. 이로부터 바이러스의 total RNA를 분리하였고 RT-PCR에 의하여 TMV pepper 계통의 외피단백질 cDNA를 합성, 증폭하였으며 이를 pBluescript II SK- 벡터에 재조합하였다. 본 실험에서 바이러스 외피단백질과 3` non-coding region을 포함하는 재조합 클론 p1561과 p1562로부터 염기서열을 분석하였고 그 결과로 477 염기의 외피단백질 유전자를 포함하는 691 염기가 합성되었음을 확인하였으며 이것과 TMV common 계통으로부터 합성된 외피단백질 cDNA와의 최대 유사도는 69%였다. 또한 유추된 아미노산 서열에서 이들 두 계통간의 최대 유사도는 81%였다.

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Quantitative Measurement of Surfactant Protein B mRNA by Filter Hybridization (Filter Hybridization 방법에 의한 Surfactant Protein B mRNA의 정량측정)

  • Park, Sung-Soo;Lee, Dong-Hoo;Shin, Dong-Ho;Lee, Jung-Hee
    • Tuberculosis and Respiratory Diseases
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    • v.39 no.3
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    • pp.242-247
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    • 1992
  • Background: The ability to precisely measure specific mRNA levels by hybridization to complementary DNA probes is an important tool for analyzing the regulation of gene expression. Surfactant proteins have important roles in regulating surfactant metabolism as well as in determing its physical properties. Method: The complete coding regions for rat surfactant protein complementary DNA of surfactant protein B were subcloned into pGem 3Z or 4Z such that either antisense or sense transcripts were obtained by using SP 6 RNA polymerase. Surfactant protein B mRNA was measured by filter hybridization. Results: Equation of standard curve between counts per minute (Y) and surfactant protein B mRNA transcript input (X) was Y=2034.9 X+159.1. Correlation coefficient was 1.0. Couclusions: Filter hybridization assay is suited to situation when rapid, accurate quantitation of multiple samples is required.

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