• Title/Summary/Keyword: hybridize

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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.

Molecular Variation and Distribution of Anopheles fluviatilis (Diptera: Culicidae) Complex in Iran

  • Naddaf, Saied Reza;Razavi, Mohammad Reza;Bahramali, Golnaz
    • Parasites, Hosts and Diseases
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    • v.48 no.3
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    • pp.231-236
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    • 2010
  • Anopheles fluviatilis James (Oiptera: Culicidae) is one of the known malaria vectors in south and southeastern Iran. Earlier ITS2 sequences analysis of specimens from Iran demonstrated only a single genotype that was identical to species Y in India, which is also the same as species T. We identified 2 haplotypes in the An. fluviatilis populations of Iran based on differences in nucleotide sequences of D3 domain of the 28S locus of ribosomal DNA (rDNA). Comparison of sequence data from 44 Iranian specimens with those publicly available in the Genbank database showed that all of the 288-D3 sequences from Kazeroun and Khesht regions in Fars Province were identical to the database entry representing species U in India. In other regions, all the individuals showed heterozygosity at the single nucleotide position, which identifies species U and T. It is argued that the 2 species may co-occur in some regions and hybridize; however, the heterozygosity in the 288-D3 locus was not reflected in ITS2 sequences and this locus for all individuals was identical to species T. This study shows that in a newly diverged species, like members of An. fluviatilis complex, a single molecular marker may not be sufficiently discriminatory to identify all the taxa over a vast geographical area. In addition, other molecular markers may provide more reliable information for species discrimination.

Molecular Coning of cDNA for Garlic Mosaic Virus Genome (마늘 모자이크 바이러스 게놈에 대한 cDNA의 클로닝)

  • 최연희
    • Journal of Plant Biology
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    • v.35 no.3
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    • pp.253-257
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    • 1992
  • Potyvirus group is the largest group among plant virus groups and damages severely plant hosts upon infectiQn. In order to investigate the mechanism by which potyviruses induce disease in plants, a cDNA clone 29-6 which is cOIlsidered to be a cDNA clone for garlic mosaic virus (GMV) was isolated. It did not hybridize to garlic latent virus genome, which is one of two major garlic viruses. Northern blot analysis shows that the genome size of garlic mosaic virus was about 9 kb. Clone 29-6 strongly hybridizes to poly(A) RNA isolated from garlic leaves, suggesting that GMV RNA is polyadenylated as other potyviruses. Nucleotide sequence analysis of cDNA clones overlapping with clone 29-6 showed that garlic plants are infected with various strains of garlic mosaic virus which are closely related to each other. other.

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Cloning and Phylogenetic Analysis of Two Different bphC Genes and bphD Gene From PCB-Degrading Bacterium, Pseudomonas sp. Strain SY5

  • Na, Kyung-Su;Kim, Seong-Jun;Kubo, Motoki;Chung, Seon-Yong
    • Journal of Microbiology and Biotechnology
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    • v.11 no.4
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    • pp.668-676
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    • 2001
  • Pseudomonas sp. strain SY5 is a PCB-degrading bacterium [24] that includes two different enzymes (BphC1 and BphC2) encoding 2,3-dihdroxybiphenyl 1,2-dioxygenase and BphD encoding 2-hydroxy-6-oxo-6-phenylhexa-2,4-dienoate hydrolase. The bphC1 and bphC2 genes were found to consist of 897 based encoding 299 amino acids and 882 bases encoding 294 amino acids, respectively, whereas the bphD gene consisted of 861 bases encoding 287 amino acids. According to a homology search, a 50% and 39% similarity between the bphC1 and bphC2 genes at the nucleotide and amino acid level was shown, respectively. The bphC1 gene showed a 38% and 45% similarity at the amino acid level to Alcaligenes eutrophus A5 and Rhodococcus rhodochrous, respectively, whereas, bphC2 showed a 95% and 43% similarity, respectively. A comparison of the deduced amino acid sequence of the bphD product of Pseudomonas sp. SY5 with that of A. eutrophus A5, Pseudomons sp. KKS102, and LB400 showed a sequence identity of 92, 92, and 79%, respectively. Strain SY5 was originally isolated from municipal sewage containing recalcitrant organic compounds an found to have a high degradability of various aromatic compounds [23]. The current study found that strain SY5 had two extradiol-type dioxygenases, which did not hybridize with each other as they had a low similarity, yet a similar structure of evolutionarily conserved amino acids residues for catalytic activity between BphC1 and BphC2 was observed.

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The Virulence Factors of Vibrio spp. (비브리오의 병원성 인자에 관한 연구)

  • Oh, Yang-Hyo;Kim, Yung-Bu;Park, Young-Min;Kim, Min-Jung;Cha, Mi-Sun;Kim, Young-Hee;Lim, Eun-Gyoung
    • The Journal of the Korean Society for Microbiology
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    • v.34 no.6
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    • pp.513-518
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    • 1999
  • A total of 113 Vibrio sp. strains were examined for plasmid content which were subjected to digestion with restriction enzymes. About the 55% Vibrio spp. have the plasmid more than one. Most of these plasmid various derivatives ranged from $2.4\;kb{\sim}23\;kb$, especially two strains of V. mimicus and one strain of V. furnissii carried one high-molecular weight plasmid (molecular weight ranging between $70\;kb{\sim}100\;kb$). Results of restriction analysis for plasmid of this three strains were by no means the rule. For detection of tdh and ctx gene, the virulence factor involved in the pathogenesis, we carried out the TDH and CT assay, PCR amplification, and hybridization. A total 11 strains were produced TDH, involved in 9 strains of V. parahaemolyticus and 1 strain of V. alginolyticus from clinical isolates and 1 strains of V. mimicus from environmental isolates. In the experiments of tdh gene detection, in all, 3 strains of V. parahaemolyticus from clinical isolates and 2 strains from environmental isolates could be successfully amplified in 400 bp by PCR. The PCR results were consistent with DNA hybridization tests. In the experiments of CT assay, in all, 3 strains of V. cholerae from clinical isolate and 1 strain of V. cholerae from environmental isolates were observed CT-producing. These CT-producing strains amplified in 302 bp by PCR for the detection of ctx gene. All CT-producing strains hybridized with digoxigenin-labeled DNA probe, while CT-negative strains did not hybridize. Also hybridization tests results for detection of ctx gene consistent with PCR.

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"All This is Indeed Brahman" Rammohun Roy and a 'Global' History of the Rights-Bearing Self

  • Banerjee, Milinda
    • Asian review of World Histories
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    • v.3 no.1
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    • pp.81-112
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    • 2015
  • This essay interrogates the category of the 'global' in the emerging domain of 'global intellectual history'. Through a case study of the Indian social-religious reformer Rammohun Roy (1772/4-1833), I argue that notions of global selfhood and rights-consciousness (which have been preoccupying concerns of recent debates in intellectual history) have multiple conceptual and practical points of origin. Thus in early colonial India a person like Rammohun Roy could invoke centuries-old Indic terms of globality (vishva, jagat, sarva, sarvabhuta, etc.), selfhood (atman/brahman), and notions of right (adhikara) to liberation/salvation (mukti/moksha) as well as late precolonial discourses on 'worldly' rights consciousness (to life, property, religious toleration) and models of participatory governance present in an Indo-Islamic society, and hybridize these with Western-origin notions of rights and liberties. Thereby Rammohun could challenge the racial and confessional assumptions of colonial authority and produce a more deterritorialized and non-sectarian idea of selfhood and governance. However, Rammohun's comparativist world-historical notions excluded other models of selfhood and globality, such as those produced by devotional Vaishnava, Shaiva, and Shakta-Tantric discourses under the influence of non-Brahmanical communities and women. Rammohun's puritan condemnation of non-Brahmanical sexual and gender relations created a homogenized and hierarchical model of globality, obscuring alternate subaltern-inflected notions of selfhood. Class, caste, and gender biases rendered Rammohun supportive of British colonial rule and distanced him from popular anti-colonial revolts and social mobility movements in India. This article argues that today's intellectual historians run the risk of repeating Rammohun's biases (or those of Hegel's Weltgeschichte) if they privilege the historicity and value of certain models of global selfhood and rights-consciousness (such as those derived from a constructed notion of the 'West' or from constructed notions of various 'elite' classicized 'cultures'), to the exclusion of models produced by disenfranchised actors across the world. Instead of operating through hierarchical assumptions about local/global polarity, intellectual historians should remain sensitive to and learn from the universalizable models of selfhood, rights, and justice produced by actors in different spatio-temporal locations and intersections.

Optimization of a microarray for fission yeast

  • Kim, Dong-Uk;Lee, Minho;Han, Sangjo;Nam, Miyoung;Lee, Sol;Lee, Jaewoong;Woo, Jihye;Kim, Dongsup;Hoe, Kwang-Lae
    • Genomics & Informatics
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    • v.17 no.3
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    • pp.28.1-28.9
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    • 2019
  • Bar-code (tag) microarrays of yeast gene-deletion collections facilitate the systematic identification of genes required for growth in any condition of interest. Anti-sense strands of amplified bar-codes hybridize with ~10,000 (5,000 each for up-and down-tags) different kinds of sense-strand probes on an array. In this study, we optimized the hybridization processes of an array for fission yeast. Compared to the first version of the array (11 ㎛, 100K) consisting of three sectors with probe pairs (perfect match and mismatch), the second version (11 ㎛, 48K) could represent ~10,000 up-/ down-tags in quadruplicate along with 1,508 negative controls in quadruplicate and a single set of 1,000 unique negative controls at random dispersed positions without mismatch pairs. For PCR, the optimal annealing temperature (maximizing yield and minimizing extra bands) was 58℃ for both tags. Intriguingly, up-tags required 3× higher amounts of blocking oligonucleotides than down-tags. A 1:1 mix ratio between up- and down-tags was satisfactory. A lower temperature (25℃) was optimal for cultivation instead of a normal temperature (30℃) because of extra temperature-sensitive mutants in a subset of the deletion library. Activation of frozen pooled cells for >1 day showed better resolution of intensity than no activation. A tag intensity analysis showed that tag(s) of 4,316 of the 4,526 strains tested were represented at least once; 3,706 strains were represented by both tags, 4,072 strains by up-tags only, and 3,950 strains by down-tags only. The results indicate that this microarray will be a powerful analytical platform for elucidating currently unknown gene functions.

Gene Expression Profiling of Human Salivary Gland Carcinogenesis with cDNA Microarray (인간 타액선 암발생에서 cDNA Microarray를 이용한 유전자발현 Profile연구)

  • Kim, Eun-Cheol;Shin, Min;Lee, Dong-Geun;Lee, Ju-Seok;Park, Myung-Hee
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.23 no.4
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    • pp.306-323
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    • 2001
  • 종양발생에서 유전자 발현을 확인하고 profile 변화를 monitor하는 것은 병리학적 변화의 원인뿐 아니라 질병탐지와 진료의 새로운 목표를 확인하기 위한 새로운 기회를 제공해준다. cDNA microarray는 수천개의 유전자 발현을 동시에 연구할 수 있는 최신의 방법으로 피부, 유방, 간을 비롯한 다른 인체장기에서는 일부 이루어졌으나 array를 이용해 타액선 종양 연구에서는 전혀 이루어지지 않았다. 인간의 타액선 세포의 악성형질전환을 조절하는 분자적 상태를 연구하기 위해 본 연구는 약 2,000개의 유전자가 print된 cDNA microarray를 이용하여 인간 타액선 도관상피세포주(HSG)와 악하선에서 기원한 미분화 선암종(SGT)간에 비교연구를 하였다. Cy3와 Cy5 dye로 각각의 세포주에서 얻은 RNA와 reciprocal hybridize시키고 GenePix 4000 scanner로 스캔하고 GenePix Pro로 분석한 후 log2로 평균발현비율을 전환시켜 최소 2배이상의 발현을 보이는 유전자를 분석대상으로 하였다. 90%이상의 유전자가 비슷한 발현을 보였으며 2배이상의 발현을 보이는 경우 HSG가 SGT에 비해 72개 유전자가, SGT가 HSG에 비해 111개의 유전자 발현이 up-regulation되어 총 10%미만의 발현차이를 보였고 반복된 hybridization 으로부터 얻은 선택된 spot의 Pearson 상관계수는 -0.85이였다. HSG에서는 6번 p 염색체에서 과발현되는 유전자가 가장 많았고, SGT에서는 11번 q 염색체에서 가장 많았는데 HSG에서는 SGT에 비해 9, 13, 17, 18, 20, 21, 22염색체에서 과발현 되는 유전자 수가 많았고, SGT에서는 HSG에 비해 2, 7, 10, 15 염색체에서 유전자 발현 증가가 관찰되었다. HSG와 SGT간의 유전 발현을 기능별로 분석한 결과 몇 가지 주요 경로가 세포악성에 관련됨을 발견하였고, 타액선 도관상피세포에서 선암종을 구별하는데 기여하는 관련된 몇종의 과다 발현된 유전자를 찾았는데 전사인자, 성장인자 및 수용기, 세포골격 및 세포외기질 단백, 세포내 신호전달조절자 및 인자, 세포표면 항원등의 그룹으로 분류할 수 있었다. 따라서 이러한 microarray를 이용한 분자학적 표지자 연구가 악성 타액선 종양 발생과정에서 큰 도움을 줄 수 있을 뿐 아니라 유전자 조절에 의한 진단, 예후, 치료에서의 정확성을 개선시킬 수 있으리라 여겨진다.

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One-Stage Polymerase Chain Reaction for the Comprehensive Detection of Type D Retrovirus Provial DNA (Type D Retrovirus 감염의 포괄적 검색을 위한 One-Stage 중합효소 연쇄반응법의 개발)

  • Jeong, Yong-Seok
    • The Journal of Korean Society of Virology
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    • v.27 no.1
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    • pp.19-27
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    • 1997
  • To develop the polymerase chain reaction (PCR) for the detection of type D simian retrovirus (SRV) infection, an oligonucleotide primer pair was designed to hybridize to the sequences within env gene of SRV subtype 1 (SRV-1). The 3' proximal env sequences annealing to the primers had been rather conserved among three different subtypes of SRV, SRV-1, SRV-2, and SRV-3 (Mason-Pfizer Monkey Virus: MPMV). The PCR using the primer pair targeting an env region successfully detected and amplified all three subtypes of SRV with excellent specificity after single round of reaction. The tests with peripheral blood mononuclear cells infected either with simian immunodeficiency virus or simian T-Iymphotropic virus type 1, major immunosuppressive viral agents together with SRV in simian, verified the specificity of the PCR by excluding any cross reactivity. Semiquantitative titration PCR, amplifying serially diluted plasmid DNA of each subtype, was performed to evaluate sensitivity limits of the reaction. Based on molecular weight of each cloned SRV genome, the PCR should be able to detect one SRV-infected cell per more than $5-7{\times}10^4$ uninfected cells after simple ethidium bromide staining of resulting products. The PCR must be very efficient screening system with its quickness, certainty, and sensitivity for SRV-infected animals used in human AIDS research model. Second round amplification of the reaction products from the first PCR, or Southern hybridization by radiolabeled probes shall render to compete its efficacy to ELISA which has been the most sensitive technique to screen SRV infection but with frequent ambiguity problem.

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Evaluation on the Performance of Coating Materials for Improving the Durability of Concretes (콘크리트의 내구성 증진을 위한 코팅재의 성능 평가)

  • Kim, Sung-Soo;Choi, Choon-Sik;Nam, Yong-Hyuk
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.7 no.4
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    • pp.99-107
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    • 2003
  • Normally coating is used a method for protecting reinforced concrete. For this purpose, organic as well as inorganic coatings are used. The advantages of inorganic coatings are lower absorption of UV, non-burning etc. On the other hand, organic coatings have the advantage of low permeability of $CO_2$, $SO_2$ and water. Organic coatings provide better protection for reinforced concrete. However, in organic coatings such as epoxy, urethane and acryl, long-term adhesive strength is reduced and the formed membrane of those is blistered by various causes. Also when organic coatings are applied to the wet surface of concrete, they have a problem with adhesion. So, we developed coating material, WGS-Eco which was hybridized with polymer and cement based material to protect concrete structures and solve problems of organic coatings. This study was conducted an comparative evaluation on physical and durable performance of developed coating material and previously used coating materials. As a result, the performance of developed coating material was not inferior to organic coating materials. So, the developed coating material was considered as a suitable coating material which had advantages of inorganic and organic coatings for protecting concrete.