• Title/Summary/Keyword: Oligonucleotide

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Detection of Soybean Mosaic Virus Using RT-PCR

  • Kim, Yul-Ho;Kim, Ok-Sun;Lee, Bong-Choon;Roh, Jae-Hwan;Kim, Myoung-Ki;Im, Dae-Joon;Hur, Il-Bong;Lee, Sang-Chul
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.44 no.3
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    • pp.253-255
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    • 1999
  • Reverse transcription and polymerase chain reaction (RT-PCR) assay was used to detect SMV strains. A pair of oligonucleotide primers were designed to include the cylindrical inclusion (CI) coding region between 4,176 to 5,560 nt. Amplification from the total RNA extracted from infected plants with SMV yielded a 1,385 bp DNA fragment. RT-PCR was shown to be $10^3$ times more sensitive than the ELISA assay and it could detect a virus in $10^{-6}$ dilution. Restriction enzyme analysis of RT- PCR products using EcoR I showed that SMV isolates were classified into six groups according to the patterns of restriction fragments.

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Fluorescence in situ hybridization using 16S rRNA-targeted oligonucleotides reveals abundance and spatial organization of methanogens in thermophilic sludge granules

  • Lee, Yu-Jin;Kim, Hyo-Seop;An, Yeong-Hui;Park, Seong-Hun
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.508-511
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    • 2000
  • In situ hybridization with fluorescent oligonucleotides(FISH) was used to detect and localize microorganisms in the granules of lab-scale upflow anaerobic sludge blanket(UASB) reactors. An UASB reactor was seeded with mesophilically-grown($35^{\circ}\;C$) granular sludge, and thermophilically($55^{\circ}\;C$) operated by feeding with a synthetic wastewater. Sections of the granules were hybridized with 16S rRNA-targeted oligonucleotide probes for Eubacteria, Archaeabacteria, and specific phylogenetic groups of methanogens. FISH clearly showed the layed structure of thermophilic granules, which was consisted of outer bacterial cells and inner archaeal cells. Methanoseata-, Methanosarcina-like cells were also found to be localized inside the granules. These results demonstrated FISH was useful in studying the spatial organizations of methanogens and in situ morphologies and metabolic functions in thermophilic granular sludges.

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Synthetic approach to the generation of antibody diversity

  • Shim, Hyunbo
    • BMB Reports
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    • v.48 no.9
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    • pp.489-494
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    • 2015
  • The in vitro antibody discovery technologies revolutionized the generation of target-specific antibodies that traditionally relied on the humoral response of immunized animals. An antibody library, a large collection of diverse, pre-constructed antibodies, can be rapidly screened using in vitro display technologies such as phage display. One of the keys to successful in vitro antibody discovery is the quality of the library diversity. Antibody diversity can be obtained either from natural B-cell sources or by the synthetic methods that combinatorially generate random nucleotide sequences. While the functionality of a natural antibody library depends largely upon the library size, various other factors can affect the quality of a synthetic antibody library, making the design and construction of synthetic antibody libraries complicated and challenging. In this review, we present various library designs and diversification methods for synthetic antibody library. From simple degenerate oligonucleotide synthesis to trinucleotide synthesis to physicochemically optimized library design, the synthetic approach is evolving beyond the simple emulation of natural antibodies, into a highly sophisticated method that is capable of producing high quality antibodies suitable for therapeutic, diagnostic, and other demanding applications. [BMB Reports 2015; 48(9): 489-494]

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

  • Kong, Hoon Young;Byun, Jonghoe
    • Biomolecules & Therapeutics
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    • v.21 no.6
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    • pp.423-434
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    • 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.

Development of Array-based Technology for Detection of HAV Using Gold-DNA Probes

  • Wan, Zhixiang;Wang, Yefu;Li, Shawn Shun-Cheng;Duan, Lianlian;Zhai, Jianxin
    • BMB Reports
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    • v.38 no.4
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    • pp.399-406
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    • 2005
  • A sensitive method for detection of Hepatitis A virus (HAV) by utilizing gold-DNA probe on an array was developed. Amino- modified oligodeoxynucleotides at the 5' position were arrayed on an activated glass surface to function as capture probes. Sandwich hybridization occurred among capture probes, the HAV amplicon, and gold nanoparticle-supported oligonucleotide probes. After a silver enhancement step, signals were detected by a standard flatbed scanner or just by naked eyes. As little as 100 fM of HAV amplicon could be detected on the array. Therefore, the array technology is an alternative to be applied in detection of HAV due to its low-cost and high-sensitivity.

Genetic Distance Study among Deoni Breed of Cattle Using Random Amplified DNA Markers

  • Appannavar, M.M.;Govindaiah, M.G.;Ramesha, K.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.3
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    • pp.315-319
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    • 2003
  • Random amplified polymorphic DNA (RAPD) analysis was done with 19 oligonucleotide primers to study genetic similarities and divergence among different types of Deoni breed of cattle viz., Balankya, Wannera and Waghya. Six random primers produced low to high numbers of polymorphic bands between pooled DNA of different Deoni types. Of the 48 RAPD markers obtained 33 were common to all Deoni types, 3 were individual specific and 12 were polymorphic for different Deoni types. The mean average percentage difference values among Deoni types showed that Balankya and Wannera had less genetic divergence when compared to Waghya.

Characterization of Graphene Oxide Suspension for Fluorescence Quenching in DNA-Diagnostics

  • Kapitonov, A.N.;Alexandrov, G.N.;Vasileva, F.D.;Smagulova, S.A.;Timofeev, V.B.;Maksimova, N.R.;Kuznetsov, A.A.
    • Korean Journal of Materials Research
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    • v.26 no.1
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    • pp.1-7
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    • 2016
  • The graphene oxides (GOs) were tested as a fluorescent quencher in the field of DNA-diagnostics. The various suspensions of GO nanoplates were prepared by changing the synthesis conditions. The suspensions were stable for at least 6 weeks by differing degrees of functionalization of various oxygen-containing groups of atoms. Depending on the properties of GO nanoplates, their fluorescent quenching abilities, which were determined by the amount of the tagged immobilized oligonucleotide, were also changed. GO suspension synthesized at $75^{\circ}C$ of reaction mixture showed the fluorescent quenching of 16.39 nmol/mg, which would be a potential substitution of molecular fluorescent quencher in test-systems for DNA-diagnostics.

Random Amplified Polymorphic DNA (RAPD) Identification of Genetic Variation in Chlorella species

  • CHO Jung Jong;KIM Yong-Tae;HUR Sung Bum;KIM Young Tae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.29 no.6
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    • pp.761-769
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    • 1996
  • The random amplified polymorphic DNA (RAPD) technique was used to characterize 18 reference strains of microalgae, mostly Chlorella species, collected from various localities around Korea peninsular. Eighteen strains consist of four genera of the family marine Chlorella from 12 samples, two genera of fresh water Chlorella from three samples, and three genera on Nannochloris. Twenty 10-mer anonymous primers were screened for amplification of genomic DNA extracted from samples using the CTAB extraction method. Nineteen of these oligonucleotide primers were positive or band producing. Three of 20 random primers (OPA 10, OPA 12, and OPA 18) resulted in both clear band and a high degree of reproducibility and showed some potential to be used to discriminate individual samples of both genetically hetero-and homogeneous populations, in determining phylogenetic relationships between species within a genus and developing individual fingerprints for each samples.

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Detection of Plasmodiophora brassicae by Using Polymerase Chain Reaction (PCR을 이용한 Plasmodiophora brassicae의 검출)

  • 지희윤;김완규;조원대;지형진;최용철
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.589-593
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    • 1998
  • DNA amplification by polymerase chain reaction (PCR) was used to specifically detect Plasmodiophora brassicae, causing clubroot of crucifers. On the basis of DNA sequence informations, an oligonucleotide primer set specific for the pathogen was designed form small subunit gene (18S-like) and internal transcribed spacer (ITS) region of ribosomal DNA. Primer ITS 5/PB-C produced an amplification product of approximately 520 bp in length with DNA from P. brassicae. However, no amplification product was produced with DNAs from several soil-borne fungi, Didymella bryoniae and Rhizopus stolonifer. Using these primers, the clubroot pathogen was readily detected from infected roots of crucifers, but not from healthy roots. Southern hybridization analysis further confirmed that the amplification product was originated from P. brassicae.

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The percutaneous absorption of antisense phosphorothioate oligonucleotide (ASPS) complementary to TGF-$\beta$ mRNA designed for scar formation inhibitor

  • Lee, Young-Mi;Lee, Sung-Hee;Kim, Su-Ung;Lee, Seong-Yong;Kim, Jaebaek;Sohn, Dong-Hwan
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1995.04a
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    • pp.129-129
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    • 1995
  • ASPS against TGF-${\beta}$ is developing as scar formation inhibitor. The scar was caused by undesired collagen deposition due to overexpression of TGF-${\beta}$ in wounded tissue. The in vitro percutaneous absorption of ASPS(25mer)was investigated by using Furanz Diffusion Cell. The flux of ASPS cannot be found through normal skin due to high molecular weight (MW 10,000) and polyanionic charge. However, the skin permeation of ASPS through tape-stripped damaged skin was markedly increased. The skin fluxs of ASPS were decreased in the following order; hairless mouse> rat >human cadaver skin.

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