• 제목/요약/키워드: fluorescent in situ hybridization

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

  • 정해원;김수영;맹승희;이용묵;유일재
    • 한국환경성돌연변이발암원학회지
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    • 제19권1호
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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.

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

  • 홍선희;안태석
    • 미생물학회지
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    • 제34권3호
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    • pp.169-174
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    • 1998
  • 소양호에서 세균군집의 계절적, 수심별 변화를 파악하고자 총세균수와 EUB338, ALG1b, BET42a, GAM42a와 CF probe 등 fluorescent rRNA target oligonuvleotide probe와 반응하는 세균 개체수를 측정하였다. 총세균수는 $0.5{\sim}2.01{\times}10^6cells{\cdot}ml^{-1}$이였으며, 2 m와 5 m 수층에서 높게 나타났다. 총세균수에 대한 Eubacteria의 비율은 22~100%이였고, Proteobacteria ${\alpha}$-group은 Eubacteria의 2.6~66.7%, ${\beta}$-group은 4.5~53.5%, ${\gamma}$-group은 4.6~76.7%, 그리고 Cytophage-Flavobacterium group은 2.1~35.9%이였다. 또한 세균군집은 계절별, 수심별로 다양한 변화를 보여, 겨울철은 ${\beta}$-group이, 봄철과 초여름은 ${\gamma}$-group이, 여름철은 ${\alpha}$-group이 우점하였고, Cytophage-Flavobacterium group이 특정적으로 우점하는 시기는 없었다. 이러한 세균 군집 구조의 분포로 계절별, 수심별로 호수에 대한 독특한 특징을 알 수 있었다.

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

  • 이영옥
    • 한국물환경학회지
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    • 제24권2호
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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.

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

  • 이영옥;조익환;김길웅
    • 유기물자원화
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    • 제8권2호
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    • pp.146-153
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    • 2000
  • 우분에 사과박을 혼합해 만든 퇴비화에서의 세균군집 특히, 질화세균의 천이를 rRNA targeted oligonucleotide probes을 사용하는 FISH(fluorescent in situ hybridization)법으로 규명하였다. 암모니아산화 세균수는 $3,3{\sim}13.4{\times}10^6cells/g$ dw의 범위에서 변화하였으며 퇴비화 26일 후에 그 최고치를 나타냈다. 반면에 아질산 산화세균은 $6.0{\sim}17.2{\times}10^6cells/g$ dw의 범위에서 변화하면서 퇴비화 7일 후에 그 최고치를 보였다. 암모니아산화세균수가 아질산산화세균수 보다 크게 나타나는 경향과 최고치를 나타내는 시점이 이들 세균군의 진정세균에 대한 비율을 측정했을 때에도 동일하였다. 암모니아산화 세균수가 아질산산화세균수보다 늦게 늦게 그 최고치를 나타내는 것은 휘발성의 암모니아가스가 퇴비화과정 초기에 고갈되었기 때문일 가능성이 크다. 아울러 본 연구결과는 FISH법이 생장이 더딘 질화세균의 검출에 유용한 도구임을 시사해 준다.

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

  • 이유진;김효섭;안영희;박성훈
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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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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    • 제38권4호
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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

  • 상병인;이윤지;김동임;김동건;김지성;유명진;박대원
    • 한국신재생에너지학회:학술대회논문집
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    • 한국신재생에너지학회 2005년도 제17회 워크샵 및 추계학술대회
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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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    • 제16권17호
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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.
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2001년도 추계학술대회
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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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Streptococcus mutans의 gtfB 및 gtfC 유전자 발현에 대한 당의 영향 (THE EFFECTS OF SUGARS ON THE EXPRESSION OF GTFB AND GTFC MRNA)

  • 정혜진;김신;정진
    • 대한소아치과학회지
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    • 제34권2호
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    • pp.299-308
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    • 2007
  • Streptococcus mutans Ingbritt의 비수용성 글루칸을 합성하는 효소인 glucosyltransferase B 및 C의 mRNA 발현에 대한 각 당의 영향을 fluorescent in situ hybridization(FISH) 방법으로 관찰하여 다음의 결과를 얻었다. 1. 1% 설탕이 함유된 BHI 배지에서 S. mutans Ingbritt를 접종하여 배양할 때 배양 9시간 째 gtfB 및 gtfC유전자의 mRNA가 발현이 급격히 증가하였다. 2 BHI 액체배지에 설탕을 첨가한 경우 gtfB및 gtfC유전자의 mRNA가 발현되었으며, 10% 설탕을 첨가한 경우 1%와 5%보다 gtfB 및 gtfC 유전자의 발현이 감소되었다. 3. 1% 설탕이 첨가된 BHI 액체 배지에 당을 첨가한 경우 포도당은 10% 첨가하였을 때 gtfB 및 gtfC 유전자의 mRNA 발현이 감소하였고, 과당의 경우 1% 첨가한 경우 감소하여 5% 와 10%에서는 gtfB및 gtfC 유전자가 거의 발현되지 않았다. 4. 자일리톨의 경우 1%부터 gtfB및 gtfC유전자 mRNA 발현이 대조군보다 감소하였고, 5%와 10%에서는 두 유전자 모두 발현이 현저하게 감소하였다. 5. 유당의 경우 두 유전자의 발현에 거의 영향을 미치지 않는 것으로 관찰되었고, 솔비톨의 경우 각 농도에서 두 유전자의 발현이 대조군보다 감소하였다. 결론적으로 비수용성 글루칸 합성하는 gtfB및 gtfC 유전자의 mRNA 발현이 포도당, 과당 자일리톨 첨가에 의해 감소함을 알 수 있었다.

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