• 제목/요약/키워드: fluorescent in situ hybridization(FISH)

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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.

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.

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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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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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.

Combined Study of Cytogenetics and Fluorescence in Situ Hybridization (FISH) Analysis in Childhood Acute Lymphoblastic Leukemia (ALL) in a Tertiary Cancer Centre in South India

  • Mazloumi, Seyed Hashem Mir;Madhumathi, D.S.;Appaji, L.;Prasannakumari, Prasannakumari
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권8호
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    • pp.3825-3827
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    • 2012
  • FISH is one of the most sensitive molecular methods to detect genetic abnormalities with DNA probes. When cytogenetic studies are normal or insufficient, FISH may detect cryptic rearrangements, rare or slowly proliferative abnormal populations in non-mitotic cells. We cytogenetically evaluated 70 childhood ALL - 67.1% were found to have an abnormal karyotype. The 23 patients (32.9%) with a normal karyotype were analyzed by FISH applying two probes; TEL/AML1 and MYB which detect cryptic rearrangements of t(12;21)(p13;q22) and deletion of (6q) respectively, associated with a good prognosis. Out of 23 patients, one was positive for t(12;21)(p13;q22) (4.3%). None of our patients were positive for MYB del(6q). Two patients showed an extra signal for MYB on chromosomes other than 6 (8.6 %) indicating amplification or duplication. Findings were compared with the available literature. Our study clearly indicated the integrated FISH screening method to increase the abnormality detection rate in a narrow range. FISH is less useful for diagnostic study of patients with suspected del(6q) but it helps in detecting known cryptic rearrangements as well as identification of new abnormalities(translocation , duplication and amplification) at the gene level.

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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Bacillus 속 세균을 검출하기 위한 Fluorescent In Situ Hybridization 방법의 개발 (Development of FISH Methods for Detection of Genus Bacillus)

  • 홍선희;김옥선;송홍규;이동훈;안태석
    • 미생물학회지
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    • 제37권3호
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    • pp.204-208
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    • 2001
  • Bacillus속에 속하는 세균을 FISH 방법으로 검출하기 위한 새로운 방법을 개발하였다. LGC353b, S-G-Bacill-0597-a-A-22와 S-G-Bacill-0597-a-S-22 등 3개의 probe를 22종의 Bacillus속 세균의 혼합배양액에 적용한 결과 S-G-Bacill-0597-a-A-22의 probe가 DAPI로 검출된 세균 중 95%가 FISH로도 측정되어 가장 효율이 좋은 것으로 나타났다. Hybrillization 과정 전에 Bacillus의 영양세포와 포자의 세포벽을 파괴하기 위하여 SDS/DTT (dithiothreitol)와 lysozyme를 순차적으로 전처리하는 방법과 lysozyme용액으로만 전처리 하는 두 가지 방법을 사용하였다. 그 결과, Bacillus를 검출하기 위해서는 영양세포의 경우에는 lysozyme용액으로만 전처리 하는 방법이 가장 좋았으며, 포자는 FISH로 검출되지 않았다. 따라서 토양, 하수 처리장등의 생태계에서 Bacillus를 검출할 경우에는 영양세포는 lysozyme으로 전처리 하여 FISH로 검출하고 포자는 따로 포자염색으로 검출하는 두가지 방법을 병행하여야한다.

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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.

gtf 유전자 발현에 대한 xylitol의 영향 (THE EFFECT OF XYLITOL ON THE EXPRESSION OF GTF GENE)

  • 염정현;정진;정태성;김신
    • 대한소아치과학회지
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    • 제31권2호
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    • pp.304-313
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    • 2004
  • 천연의 당알콜계 감미료인 자일리톨을 사용하여 자당으로부터 글루캔을 합성하는 효소인 glucosyltransferase(GTF)의 mRNA 발현에 미치는 영향을 Fluorescent in situ hybridization(FISH)과 유식세포측정기를 이용하여 분석하여 다음의 결과를 얻었다. 1. FISH분석결과 1% 자당이 함유된 BHI 배지에 1%, 5%, 10% 자일리톨을 첨가하였을 때 gtfB, gtfC 및 gtfD mRNA 발현이 농도 의존적으로 억제되었고, gtfB 보다는 gtfC 및 gtfD의 발현이 더 크게 억제되었다. 2. 유식세포측정기로 분석한 결과 자당의 농도가 0.1%, 0.5%, 1%로 증가함에 따라 gtfB, gtfC 및 gtfD mRNA 발현이 증가하였다. 3. 유식세포측정기로 분석한 결과 자당이 1%함유되었을 때 1%, 5%, 10% 자일리톨을 첨가한 경우 gtfB, gtfC 및 gtfD mRNA발현이 유사하게 억제되었으며, 10% 자일리 톨을 첨가한 경우 gtfB, gtfC 및 gtfD mRNA발현이 가장 크게 억제되었다. 이상의 결과를 종합하면 자일리톨은 자당첨가에 의한 Streptococcus mutars의 gtf mRNA 발현을 억제하여 항우식효과에 기여하는 것으로 생각된다.

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