• Title/Summary/Keyword: fluorescent in situ hybridization

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Detection of Chromosomal Rearrangements by Chromium in Human Lymphocyte Using Fluorescence in situ Hybridization (FISH) with Triple Combination of Composite whole Chromosome Specific Probe (FISH(fluorescence in situ hybridization)를 이용하여 분석한 크롬에 의해 유발된 염색체 이상)

  • 정해원;김수영;맹승희;이용묵;유일재
    • Environmental Mutagens and Carcinogens
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    • v.19 no.1
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    • pp.14-19
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    • 1999
  • Chromosome rearrangements induced in human lymphocyte after in vitro exposure to chromium were analysed by the use of fluorescence in situ hybridization(FISH) with triple combination of composite whole chromosome-specific probe for chromosome 1, 2 and 4. Chromosome aberrations was scored by the Protocol for Aberration Identification and Nomenclature Terminology (PAINT). Stable translocation was the most frequent type of aberrations and dicentrics and insertions were also observed. Chromium treatment enhanced the frequencies of stable translocations and color junctions in a dose-dependent manners, but no distinct increase of dicentrics and insertions was seen. The ratio of the yields of translocation to the yields of dicentric varied between 13 to 27. The presents results demonstrate fluorescent in situ hybridization (FISH) is useful for detecting chromosomal rearrangements induced by chromium.

Seasonal Changes of bacterial community analysed by fluorescent in situ hybridization method in Lake Soyang (Fluorescent In Situ Hybridization방법으로 분석한 소양호 세균 군집 구조의 계절적 변화)

  • Hong, Sun-Hee;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.169-174
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    • 1998
  • To define the structure and diversity of bacterial communities in the aqutic ecosystem, Lake Soyang, the largest artificial reservoir in Korea, a new method, fluorescent in situ hybridization was applied. This technique relies on the specific hybridization of the nucleic acid probes to the naturally amplified intracellular rRNA. By this method, the bacterial community composition of Lake Soyang and bacterial numbers belong to eubacteria, proteobacteria and Cytophaga-Flavobacterium group were estimated. Total bacterial numbers ranged from $0.3{\times}10^6{\sim}2.0{\times}10^6cells{\cdot}ml^{-1}$, and vertical profile of total bacteria showed the peak at 2 and 5 m depths. The ratio of eubacteria to total bacteria were 22~100% and varied with depth and season. The percentage of Proteobacteria ${\alpha}$-group ranged 2.6~66.7%, ${\beta}$-group 4.5~53.5%, ${\gamma}$-group 4.6~76.7% and Cytophaga-Flavobacterium group 2.1~35.9%. Also, bacteria] community had spatial and temporal characteristics. The dominant groups were ${\beta}$-group in winter, ${\gamma}$-group in spring and early summer and ${\alpha}$-group in summer.

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Changes of Nitrifying Bacteria in the Different Zone (Upper·Mid·Lower Part) of the Nak-Dong River (낙동강 상·중·하 수역에서의 질화세균군의 변화)

  • Lee, Young-Ok
    • Journal of Korean Society on Water Environment
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    • v.24 no.2
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    • pp.214-220
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    • 2008
  • Nitrifying bacteria were detected by fluorescent in situ hybridization (FISH) method at 6 sampling sites with different eutrophication degree in the Nak-Dong River and their tributaries. And conventional physico-chemical parameters including $NH_4-N$, $NO_3-N$, and TN were determined concurrently. In rainy period (July), there was no noticeable difference between the number of ammonia/nitrite-oxidizing bacteria detected at each site except Sang-Ju and the ratio of nitrifying bacteria to total counts stained by DAPI varied in 6~33%. By contrast, in the dry period (October), both of bacterial population was increased differently and the ratio of nitrifying bacteria to total counts ranged more widely from 6% in heavily polluted water zone, Hwa-Won to 60% in upper tributary with high agricultural land use. Byung-Sung-Chun. In January, the numbers of ammonia-oxidizing bacteria was reduced up to one tenth, while those of nitrite-oxidizing bacteria was apparently increased maybe due to high DO and low DOC.

Succession of Bacterial Populations in Cattle Manure Compost as Determined by Fluorescent In Situ Hybridization (우분 퇴비화에서의 Fluorescent In Situ Hybridization법에 의한 세균군집의 천이)

  • Lee, Young-Ok;Jo, Ik-Hwan;Kim, Kil-Woong
    • Journal of the Korea Organic Resources Recycling Association
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    • v.8 no.2
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    • pp.146-153
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    • 2000
  • To elucidate succession of bacterial populations, especially nitrifying bacteria during the composting of cattle manure with apple pomace, fluorescent in situ hybridization(FISH) using rRNA targeted oligonucleotide probes were applied. The density of ammonia-oxidizing bacteria was ranged from $3,3{\times}10^6cells/g$ dw to $13,4{\times}10^6cells/g$ dw with the peak value after 26 composting days whereas that of nitrite-oxidizing bacteria varied between $6.0{\times}10^6cells/g$ dw and $17.2{\times}10^6cells/g$ dw with the peak value after 7 composting days. And the tendency that the numbers of nitrite-oxidizing bacteria were higher than those of ammonia-oxidizing bacteria, and the peak-time of their densities were the same as that of data determined by the ratio of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria to eubacteria. The peak of ammonia-oxidizing bacteria followed the peak of nitrite-oxidizing bacteria, at the late phase of composting process could be probably caused by the depletion of volatile ammonia of composting materials. Besides these results indicate that FISH method is a useful tool for detection of slow growing nitrifying bacteria.

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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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Duplication of intrachromosomal insertion segments $4q32{\rightarrow}q35$ confirmed by comparative genomic hybridization and fluorescent $in$ $situ$ hybridization

  • Kim, Jin-Woo;Park, Ju-Yeon;Oh, Ah-Rum;Choi, Eun-Young;Ryu, Hyun-Mee;Kang, Inn-Soo;Koong, Mi-Kyoung;Park, So-Yeon
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.4
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    • pp.238-241
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    • 2011
  • A 35-year-old man with infertility was referred for chromosomal analysis. In routine cytogenetic analysis, the patient was seen to have additional material of unknown origin on the terminal region of the short arm of chromosome 4. To determine the origin of the unknown material, we carried out high-resolution banding, comparative genomic hybridization (CGH), and FISH. CGH showed a gain of signal on the region of $4q32{\rightarrow}q35$. FISH using whole chromosome painting and subtelomeric region probes for chromosome 4 confirmed the aberrant chromosome as an intrachromosomal insertion duplication of $4q32{\rightarrow}q35$. Duplication often leads to some phenotypic abnormalities; however, our patient showed an almost normal phenotype except for congenital dysfunction in spermatogenesis.

음식물쓰레기와 폐활성 슬러지를 이용한 생물학적 수소생산

  • Sang, Byeong-In;Lee, Yun-Ji;Kim, Dong-Im;Kim, Dong-Geon;Kim, Ji-Seong;Yu, Myeong-Jin;Park, Dae-Won
    • 한국신재생에너지학회:학술대회논문집
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    • 2005.11a
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    • pp.299-306
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    • 2005
  • Anaerobic fermentation of food waste (FW) and waste activated sludge (WAS) for hydrogen production was performed in CSTR (Continuous Stirred tank reactor) under various HRTs and volumetric mixing ratio (V/V) of two substrates, FW and WAS. The specific hydrogen production potential of FW was higher than that of WAS. However, pH drop in the CSTR for hydrogen production from FW was higher than that from WAS. The maintenance of desired pH during fermentative hydrogen production is regarded as the most important operation parameter for the stable hydrogen production. Therefore, when the potential of hydrogen production from FW and better buffer capacity of WAS, the proper mixture of FW and WAS for fermentative hydrogen production were considered as a useful complementary substrate. The maximum yield of specific hydrogen production, 140 mL/g VSS, was found at HRT of 2 day and the volumetric mixing ratio of 20:80 (WAS : FW). The spatial distribution of hydrogen producing bacteria was observed in anaerobic fermentative reactor using fluorescent in situ hybridization (FISH) method.

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Detection of Gene Amplification by Multiplex Ligation-Dependent Probe Amplification in Comparison with In Situ Hybridization and Immunohistochemistry

  • Tabarestani, Sanaz;Ghaderian, Sayyed Mohammad Hossein;Rezvani, Hamid
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.17
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    • pp.7997-8002
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    • 2015
  • Gene amplification is an important mechanism in the development and progression of cancer. Currently, gene amplification status is generally determined by in situ hybridization (ISH). Multiplex ligation-dependent probe amplification (MLPA) is a PCR-based method that allows copy number detection of up to 50 nucleic acid sequences in one reaction. The aim of the present study was to compare results for HER2, CCND1, MYC and ESR1 gene amplification detected by MLPA with fluorescent in situ hybridization (FISH) and chromogenic in situ hybridization (CISH) as clinically approved methods. Tissue samples of 170 invasive breast cancers were collected. All were ER positive. Tissue samples had previously been tested for HER2 using immunohistochemistry. Amplification of the selected genes were assessed using MLPA, FISH and CISH and results were compared. HER2 MLPA and ISH results were also compared with HER2 immunohistochemistry (IHC) which detects protein overexpression. Amplification of HER2, CCND1, MYC and ESR1 by MLPA were found in 9%, 19%, 20% and 2% of samples, respectively. Amplification of HER2, CCND1, MYC and ESR1 by FISH was noted in 7%, 16%, 16% and 1% of samples, respectively. A high level of concordance was found between MLPA/FISH (HER2: 88%, CCND1: 88%, MYC: 86%, ESR1: 92%) and MLPA/CISH (HER2: 84%). Of all IHC 3+ cases, 91% were amplified by MLPA. In IHC 2+ group, 31% were MLPA amplified. In IHC 1+ group, 2% were MLPA amplified. None of the IHC 0 cases were amplified by MLPA. Our results indicate that there is a good correlation between MLPA, IHC and ISH results. Therefore, MLPA can serve as an alternative to ISH for detection of gene amplification.

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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THE EFFECTS OF SUGARS ON THE EXPRESSION OF GTFB AND GTFC MRNA (Streptococcus mutans의 gtfB 및 gtfC 유전자 발현에 대한 당의 영향)

  • Chung, Hye-Jin;Kim, Shin;Chung, Jin
    • Journal of the korean academy of Pediatric Dentistry
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    • v.34 no.2
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    • pp.299-308
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    • 2007
  • Insoluble glucan is the important component of oral biofilm, which is synthesized from sucrose through the action of glucosyltransferase (GTF) B and GTF C encoded by the gtfB and gtfC genes, respectively of Streptococcus mutans. In present study, the effects of various sugars on the mRNA expression of gtfB and gtfC of S. mutans Ingbritt were examined by fluorescent in situ hybridization (FISH). The mRNA of gtfB and gtfC was expressed normally in the BHI broth containing 1% and 5% sucrose. The mRNA expression was decreased by the addition of 10% of glucose, and 1%, 5% and 10% of fructose. Lactose had no great effect on the expression of gtfB and gtfC. 5% and 10% of xylitol greatly reduced the mRNA expression of gtfB and gtfC. Sorbitol slightly decreased the mRNA expression of gtfB and gtfC when compared to the control. In summary, mRNA expression of gtfB and gtfC was decreased by the addition of glucose, fructose, and xylitol.

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