• Title/Summary/Keyword: hybridization

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Histopathologic Characterization of Viral Pathogens in Cultured Olive Flounder, Paralichthys Olivaceus, using in-situ Hybridization Methods (In-situ hybridization 법을 사용한 양식 넙치, Paralichthys olivaceus의 바이러스 감염 질병 특성 고찰)

  • Do, Jeong Wan;Lee, Nam-Sil;Jung, Sung Hee;Kim, Kyung-Kil;Choi, Hye Sung;Park, Jeong Woo;Kim, Yi Cheong
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.163-171
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    • 2013
  • Polymerase chain reaction (PCR) is the most rapid and widely used method to detect viral pathogens. However, this method does not provide histopathologic nature of the virus. In situ hybridization (ISH) with oligonucleotide probes is attractive because it is a rapid method for detection and identification of viral pathogens at sites of tissue infection. In order to understand the histopathologic characterictics of Red sea bream iridovirus (RSIV), viral-hemorrhagic septicemia (VHS) virus and viral nervous necrosis (VNN) virus to cultured olive flounder, we her applied ISH method to various kinds of olive flounder tissues with PCR-positive for these three viruses. We found that these viruses showed different tissue tropism and were detected from different cell types. Our results suggest that ISH is useful not only in rapid detection of viral pathogens but also in understanding the histopathologic characters of specific viral pathogens.

Production and Characterizations of Somatic Hybrids between Brassica campestris L. ssp pekinensis and Brassica of oleracea L. var capitata

  • Lian, Yu-Ji;Lim, Hak-Tae
    • Journal of Plant Biotechnology
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    • v.3 no.1
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    • pp.33-38
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    • 2001
  • Protoplasts isolated from inbred lines of Brassica oleracea L. var capitata (cabbage) and Brassica campestris L. ssp. pekinensis (Chinese cabbage) were fused by PEG-mediated method, and somatic hybrid cells were differentiated into plants. for the identification of somatic hybrid plants, ploidy level, plant morphology, and cytological analysis were performed. All of the regenerated plants derived from fused protoplasts were shown to be 2X-4X, or higher ploidy level, presumably due to somatic hybridization or chromosome doubling. The morphology of leaves, petioles, and flowers showed an intermediate phenotype between Chinese cabbage and cabbage. Chromosome numbers in these somatic hybrids ranged mostly from 33 to 38. According to Genomic in situ hybridization (GISH) pattern, signals from both fusion parents of B.campestris or B.oleracea were detected in different colors when chromosomes of putative somatic hybrids were observed.

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Effects of Ursolic Acid on Oncogene Expression Detected by In Situ Hybridization in Mice (생쥐에서 종양세포의 암유전자발현에 대한 울솔산의 효과)

  • Rhew, Tae-Hyong;Park, Sung-Mi;Park, Kun-Young;Chung, Hae-Young;Hah, Jae-Chung;Lee, Chung-Kyu
    • YAKHAK HOEJI
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    • v.36 no.6
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    • pp.529-537
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    • 1992
  • To investigate the effect of ursolic acid on the expression of oncogenes in tumor cells of mice, sarcoma 180 ascites tumor cells were implanted into the left groin of ICR mice and the tumor bearing mice were treated with ursolic acid. The expression of oncogenes were measured by in situ hybridization method. Ursolic acid significantly reduced the expression of oncogenes in the tumor cells. Therefore, it can be said that the prestated anticarcinogenic effect of ursolic acid could be partly ascribed to the mechanism included in the oncogene´s transcription level.

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Analysis of Mitochondrial DNA Mutation in hepatoma

  • Chung, Ku-Sun;Lee, Kyo-Young;Shim, Sang-In;Kim, Jin-Sun;Song, Eun-Sook
    • BMB Reports
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    • v.33 no.5
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    • pp.417-421
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    • 2000
  • Mitochondrial DNA (mtDNA) mutation was investigated in a hepatoma patient using a polymerase chain reaction (PCR) and an in situ hybridization technique. Biotin-labeled probes for the subunit m of cytochrome c oxidase revealed differences in the in situ hybridization. A PCR assay using biopsied and microdissected tissues showed that common deletion (4,977 bp) was more pronounced in the cancer region than in the normal parts of the same patient. These results suggest that mtDNA deletion might be associated with tumorigenesis in hepatoma.

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Sex Determination of In Vitro Fertilized Bovine Embryos by Fluorescence in situ Hybridization Technique

  • Han, M. S.;E. J. Cho;H. B. Ha;Park, H. S.;S. H. Sohn
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.287-287
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    • 2004
  • Sexing from bovine embryos which were fertilized in vitro implicate a possibility of production of the sex controlled cattle. This study was carried out to investigate the possibility of determining of embryo sex by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe which constructed from the btDYZ-1 sequences. (omitted)

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Detection of Waterborne Pathogens by PCR-reverse Blot Hybridization

  • Choi, Yeon-Im;Lee, Gyu-Sang;Bang, Hye-Eun;Kim, Jong-Bae;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.16 no.1
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    • pp.10-18
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    • 2010
  • The present study was set to develop comprehensive system for assessing the safety of drinking water using PCR-reverse blot hybridization assay (REBA). The REBA developed in this study can detect waterborne pathogens such as Shigella spp., Salmonella spp., Citrobacter spp., Enterobacter spp., Klebsiella spp., Yersinia spp., Mycobacterium spp., Listeria spp. at the genus level, and Escherichia coli, Citrobacter freundii, Klebsiella pneumoniae, Pseudomonas aeruginosa, Yersinia enterocolitica, Y. pseudotuberculosis, Mycobacterium avium complex, M. marinum, Enterococcus faecalis, and Staphylococcus aureus at the species level, and E. coli O157:H7 at the strain level.

In Situ Identification of Cyanobacteria

  • Ahn Tae-Seok;Hong Sun-Hee;Chung Hyun-Mi;Belkova Natalia L.
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2001.11a
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    • pp.121-127
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    • 2001
  • Seven cyanobacteria strains (Anabaena macrospora NIERl0016, Oscillatoria sp. NIER10042, Microcystis aeruginosa NIER10015, M. ichtyoblabe BIER10025, BIER10040, M. novacekii NIER10029, M. wesenbergii NIER10068) were tested with four rRNA - targeted oligonucleotide probes labelled with horseradish peroxidase (HRP) and specific for cyanobacteria. Non- fluorescent detection of hybridization signal was used. The hybridization with artificial mixture of cyanobacteria have shown the possibility to use 2 species-specific probes in duplicate hybridization and detection with different colored substrates.

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Radiation induced Chromosome aberration in human Iymphocyte detected by Fluorescence in sifu hybridization (FISH(Fluorescence in situ hybridization)를 이용하여 분석한 방사선에 의해 유발된 림프구의 염색체 이상)

  • 정해원;손은희;기혜성;하성환
    • Environmental Mutagens and Carcinogens
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    • v.16 no.2
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    • pp.88-96
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    • 1996
  • Fluorescence in situ hybridization (FISH) with the DNA probe for human chromosome 4 was used to analyse in vitro radiation induced chromosome rearrangement in peripheral lymphocyte. Translocations, dicentrics, acentrics and color junctions involving the painted chromosome were scored according to the Protocol for Aberration Identification and Nomenclature Terminology (PAINT) system. The frequency of chromosome rearrangements including reciprocal translocation, dicentric, acentric fragment and color junction increased with radiation dose. The frequency of dicentric chromosome reduced by the fixation time following irradiation, whereas that of translocation was relatively persistent. The applicability of FISH for scoring stable translocation for biological dosimetry was demonstrated.

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Characterization of denaturation and renaturation of DNA for DNA hybridization

  • Wang, Xiaofang;Lim, Hyun Jeong;Son, Ahjeong
    • Environmental Analysis Health and Toxicology
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    • v.29
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    • pp.7.1-7.8
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    • 2014
  • Objectives The present study was designed to systematically characterize the denaturation and the renaturation of double stranded DNA (dsDNA), which is suitable for DNA hybridization. Methods A series of physical and chemical denaturation methods were implemented on well-defined 86-bp dsDNA fragment. The degree of each denaturation was measured and the most suitable denaturation method was determined. DNA renaturation tendency was also investigated for the suggested denaturation method. Results Heating, beads mill, and sonication bath did not show any denaturation for 30 minutes. However probe sonication fully denatured DNA in 5 minutes. 1 mol/L sodium hydroxide (alkaline treatment) and 60% dimethyl sulfoxide (DMSO) treatment fully denatured DNA in 2-5 minutes. Conclusions Among all the physical methods applied, the direct probe sonication was the most effective way to denature the DNA fragments. Among chemical methods, 60% DMSO was the most adequate denaturation method since it does not cause full renaturation during DNA hybridization.

Identification of Salmonella pullorum Genomic Sequences Using Suppression Subtractive Hybridization

  • Li, Qiuchun;Xu, Yaohui;Jiao, Xinan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.9
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    • pp.898-903
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    • 2009
  • Pullorum disease affecting poultry is caused by Salmonella enterica serovar Pullorum and results in severe economic loss every year, especially in countries with a developing poultry industry. The pathogenesis of S. Pullorum is not yet well defined, as the specific virulence factors still need to be identified. Thus, to isolate specific DNA fragments belonging to S. Pullorum, this study used suppression subtractive hybridization. As such, the genome of the S. Pullorum C79-13 strain was subtracted from the genome of Salmonella enterica serovar Gallinarum 9 and Salmonella enterica serovar Enteritidis CMCC(B) 50041, respectively, resulting in the identification of 20 subtracted fragments. A sequence homology analysis then revealed three types of fragment: phage sequences, plasmid sequences, and sequences with an unknown function. As a result, several important virulence-related genes encoding the IpaJ protein, colicin Y, tailspike protein, excisionase, and Rhs protein were identified that may play a role in the pathogenesis of S. Pullorum.