• Title/Summary/Keyword: Fluorescence In situ Hybridization

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Performance and Microbial Characteristics of Bio-hydrogen Production from Food Waste with Thermal Pre-treatment (음식물류 폐기물의 혐기성 수소 발효 시 열처리에 따른 성능 및 미생물 특성 평가)

  • Lee, Chaeyoung;Choi, Jaemin
    • Transactions of the Korean hydrogen and new energy society
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    • v.25 no.1
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    • pp.11-18
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    • 2014
  • This study was conducted to investigate the effect of thermal pre-treatment on bio-hydrogen from food waste. Two continuous reactors operated and VFAs(volatile fatty acids) production and microbial communities were analyzed. The average hydrogen yield was 0.50 and 0.33mol $H_2/mol$ $hexose_{added}$ in thermally treated food added reactor(R1) and control(R2), respectively. Butyrate concentration was similarly 7,500mg/L in both reactors, but two times higher lactate concentration was observed in R2(3,800mg/L). The results of FISH(fluorescence in situ hybridization) showed that the relative microorganism to hydrogen producing bacteria was 78 and 27% in R1 and R2, respectively.

Karyotype Analyses of a Rice Cultivar 'Nakdong' and its Four Genetically Modified Events by Conventional Staining and Fluorescence in situ Hybridization

  • Jeon, Eun Jin;Ryu, Kwang Bok;Kim, Hyun Hee
    • Korean Journal of Breeding Science
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    • v.43 no.4
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    • pp.252-259
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    • 2011
  • Conventional staining and fluorescence in situ hybridization (FISH) karyotypes of the non-genetically modified (GM) parental rice line, 'Nakdong' (Oryza sativa L. japonica), and its four GM rice lines, LS28 (event LS30-32-20-1), Cry1Ac1 (event C7-1-9-1), and LS28 ${\times}$ Cry1Ac1 (events L/C1-1-3-1 and L/C1-3-1-1) were analyzed using 5S and 45S rDNAs as probes. Both parental and transgenic lines were diploids (2n=24) with one satellite chromosome pair. The lengths of the prometaphase chromosomes ranged from 1.50 to $6.30{\mu}m$. Four submetacentric and eight metacentric pairs comprised the karyotype of 'Nakdong' and its four GM lines. One pair of 5S rDNA signals was detected near the centromeric region of chromosome g in both the parental and transgenic lines. The 45S rDNA signals were detected on the secondary constrictions of the satellite chromosome pair in both the parental and transgenic lines. There was no significant difference in chromosome size, length, and composition between 'Nakdong' and its four GM lines. This research was conducted as a preliminary study for chromosomal detection of transgenes in GM rice lines and would be useful for their breeding programs.

FISH기법 적용을 위한 Y 염색체 특이 DNA Probe의 개발

  • 조은정;류란숙;류은경;손시환
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.24-24
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    • 2003
  • Fluorescence in situ Hybridization(FISH)는 특정 염기서열을 이용하여 염색체나 염색체상의 DNA위치를 확인하는 기술로서, 면역세포화학 기술과 결합되어져 현미경으로 이들의 유전적 활성도를 직접 확인할 수 있는 방법으로 지금까지의 radioisotopes 대신 non-radioactive labeling 방법으로서 fluorescence을 이용한 분자세포유전학적 검정 방법이다. 따라서 특정 염색체의 FISH probe의 개발은 FISH 기법을 이용하여 조직 또는 세포내 특정 염색체나 DNA의 존재나 이상 유무를 신속하고 정확하게 파악할 수 있다. 본 연구는 소와 사람을 대상으로 Y-염색체 특이 DNA probe를 개발하고 이를 이용하여 FISH를 시행함으로서 본 probe의 신뢰성을 확인하고 임상적 적용 가능성을 제시 하고자 하였다.

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Deciphering Functions of Uncultured Microorganisms (난배양성 미생물의 기능 분석 방법)

  • Kim, Jeong-Myeong;Song, Sae-Mi;Jeon, Che-Ok
    • Korean Journal of Microbiology
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    • v.45 no.1
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    • pp.1-9
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    • 2009
  • Microbes within complex communities show quite different physiology from pure cultured microbes. However, historically the study of microbes has focused on single species in pure culture and most of microbes are unculturable in our labs, so understanding of complex communities lags behind understanding of pure cultured cells. Methodologies including stable isotope probing (SIP), a combination of fluorescence in situ hybridization (FISH) and microautoradiography (MAR), isotope micrarray, and metagenomics have given insights into the uncultivated majority to link phylogenetic and functional information. Here, we review some of the most recent literatures, with an emphasis on methodological improvements to the sensitivity and utilities of these methods to link phylogeny and function in complex microbial communities.

Microbial and Physicochemical Monitoring of Granular Sludge During Start-up of Thermophilic UASB Reactor

  • Ahn, Yeong-Hee;Park, Sung-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.13 no.3
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    • pp.378-384
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    • 2003
  • Mesophilically-grown granular sludge seeded in thermophilic UASB reactor was monitored to better understand the start-up process of the reactor. The reactor was fed with a synthetic wastewater containing glucose. As COD loading rate increased stepwise, methane production rate increased. Maximum values of COD removal efficiency (95%) and methane production rate (5.3 l/day) were achieved by approximately day-80 and remained constant afterward. However, physicochemical and microbial properties of granules kept changing even after day-80. Specific methanogenic activity (SMA) was initially negligible, and increased continuously until day-153 and remained constant afterward, showing the maximum value of $1.51{\pm}0.13\;g\;CH_4-COD/g$ VSS/day. Deteriorated settling ability of granules recovered the initial value by day-98 and was maintained afterward, as determined by sludge volume index. Initially reduced granule size increased until day-126, reaching a plateau of 1.1 mm. Combined use of fluorescence in situ hybridization and confocal laser scanning microscopy (CLSM) allowed to localize families of Methanosaetaceae and Merhanosarcinaceae in granules with time Quantitative analyses of CLSM images of granule sections showed abundance patterns of the methanogens and numerical dominance of Methanosaeta spp. throughout the start-up period. The trend of SMA agreed well with abundance patterns of the methanogens.

Chromosome Analysis Using GISH and FISH of Interspecific Hybrids between Allium cepa L. and A. fistulosum L. (GISH와 FISH를 이용한 양파와 파간 종간교잡계통의 염색체 분석)

  • Kim, Cheol-Woo;Lee, Eul-Tai;Kim, Hwa-young;Choi, In-Hu;Bang, Jin-Ki;Koo, Dal-Hoe;Bang, Jae-Wook
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.468-473
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    • 2009
  • Fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) were used for chromosome analysis of hybrids (2n=16) between onion (Allium cepa L., 2n=2X=16) and welsh onion (A. fistulosum L., 2n=2X=16). 5S rDNA, 45S rDNA, and tandemly repeated DNA (TSD) sequence were used as probes for FISH analysis. A. fistulosum specific DNA probe of telomeric repeats and A. fistulosum DNA were used for GISH analysis. In the analysis of meiotic chromosome GISH revealed that hybrids have 7 bivalants and 2 univalents chromosome and 2 univalents were derived from A. fistulosum chromosomes. In somatic chromosomes of hybrid each 8 chromosomes were derived from A. cepa and A. fistulosum, respectively. FISH signal of 45S rDNA probe in A. fistulosum was detected at secondary constriction of chromosomes, while FISH signal in A. cepa was observed in both secondary constriction and telomere of chromosomes. TDS signals in A. fistulosum chromosomes were detected at all subtelomeric of 8 chromosomes and also in 2 pericentromeric of the chromosomes, whereas TDS signals in A. cepa were observed only in subtelomeric in all chromosomes. The pattern of TDS signal in hybrid chromosomes was similar to those of A. fistulosum chromosomes.

Rapid Sex Identification of Chicken by Fluorescence In Situ Hybridization Using a W Chromosome-specific DNA Probe

  • Sohn, S.H.;Lee, C.Y.;Ryu, E.K.;Han, J.Y.;Multani, A.S.;Pathak, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.11
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    • pp.1531-1535
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    • 2002
  • It has been known that the sex of chicken cells can be most accurately identified by fluorescence in situ hybridization (FISH). However, the presently available FISH has not been widely used for sex identification, because the procedures for cell preparation and FISH itself are complicated and time-consuming. The present study was undertaken to test a rapid FISH procedure for sexing chicken. A FISH probe was simultaneously synthesized and labeled with digoxigenin by polymerase chain reaction (PCR) targeting a 416 bp segment of the 717 bp XhoI family fragment which is repeated over 10 thousand times exclusively in the W chromosome. Sexing by FISH was performed on cytological preparations of early embryos, adult lymphocytes and feather pulps of newly hatched chicks. The DNA probe hybridized to all types of uncultured interphase as well as metaphase female but not male cells that had been examined. Moreover, consistent with the known site of the XhoI family, the hybridization signal was localized to the pericentromeric region of the W chromosome. We, therefore, conclude that the present PCR-based FISH can be used as a rapid and reliable sex identification procedure for chicken.

Genome-wide Examination of Chromosomal Aberrations in Neuroblastoma SH-SY5Y Cells by Array-based Comparative Genomic Hybridization

  • Do, Jin Hwan;Kim, In Su;Park, Tae-Kyu;Choi, Dong-Kug
    • Molecules and Cells
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    • v.24 no.1
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    • pp.105-112
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    • 2007
  • Most neuroblastoma cells have chromosomal aberrations such as gains, losses, amplifications and deletions of DNA. Conventional approaches like fluorescence in situ hybridization (FISH) or metaphase comparative genomic hybridization (CGH) can detect chromosomal aberrations, but their resolution is low. In this study we used array-based comparative genomic hybridization to identify the chromosomal aberrations in human neuroblastoma SH-SY5Y cells. The DNA microarray consisting of 4000 bacterial artificial chromosome (BAC) clones was able to detect chromosomal regions with aberrations. The SH-SY5Y cells showed chromosomal gains in 1q12~ q44 (Chr1:142188905-246084832), 7 (over the whole chro-mosome), 2p25.3~p16.3 (Chr2:18179-47899074), and 17q 21.32~q25.3 (Chr17:42153031-78607159), while chromosomal losses detected were the distal deletion of 1p36.33 (Chr1:552910-563807), 14q21.1~q21.3 (Chr14:37666271-47282550), and 22q13.1~q13.2 (Chr22:36885764-4190 7123). Except for the gain in 17q21 and the loss in 1p36, the other regions of gain or loss in SH-SY5Y cells were newly identified.

Characteristics of Microbial Distribution of Nitrifiers and Nitrogen Removal in Membrane Bioreactor by Fluorescence in situ Hybridization (막/생물반응기에서 Fluorescence in situ Hybridization 기법을 이용한 질산화 미생물 분포특성 및 질소제거 연구)

  • Lim Kyoung-Jo;Kim Sun-Hee;Kim Dong-Jin;Cha Gi-Cheol;Yoo Ik-Keun
    • Microbiology and Biotechnology Letters
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    • v.34 no.3
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    • pp.257-264
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    • 2006
  • An aerobic submerged membrane bioreactor (MBR) treating ammonium wastewater was studied in respect of nitrification characteristics and distribution of nitrification bacteria over a period of 350 days. MBR was fed with ammonium concentration of 500-1000 mg $NH_4-N/L$ at a nitrogen load of $1-2kg\;N/m^3{\cdot}d$. Overall ammonium oxidation rate increased with dissolved oxygen (DO) concentration, temperature, and sludge retention time (SRT). Under a higher concentration of free ammonia ($NH_3-N$) due to the decrease of ammonium oxidation rate, the nitrite ratio ($NO_2-N/NO_x-N$) in the effluent increased. The sudden collapse of nitrification efficiency accompanied by sludge foaming and the increase of sludge volume index (SVI) was observed unexpectedly during the operation. At the later stage of operation, additional carbon source was fed to the MBR and resulted in twice higher value of SVI and the decrease of ammonium oxidation rate. In fluorescence in situ hybridization (FISH) analysis, genus Nitrosomonas which is specifically hybridized with probe NSM156 was initially the dominant ammonia oxidizing bacteria and the amount of Nitrosospira gradually increased. Nitrospira was the dominant nitrite oxidizing bacteria during whole operational period. Significant amount of Nitrobacter was also detected which might due to the high concentration of nitrite maintained in the reactor.

Relationship between the DNA content of human chromosome and their contribution to radiation-induced chromosome aberration analysed by fluorescence in situ hybridization(FISH) (방사선에 의한 염색체이상과 DNA 함량과의 관계)

  • Chung, Hai-Won;Kim, Su-Young;Ha, Sung-Whan;Kim, Tae-Hwan;Cho, Chul-Koo
    • Journal of Radiation Protection and Research
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    • v.26 no.2
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    • pp.101-111
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    • 2001
  • To study the relationship between the DNA content of human chromosomes and their involvement in radiation-induced structural chromosome aberrations, the frequencies of translocations and dicentrics induced in human lymphocytes after in vitro exposure to 2Gy of x-tay were analysed by fluorescence in situ hybridization(FISH). Single whole chromosome probes(WCP), specific for chromosomes 1, 2, 4, 7, 8, 9, and 21 and triple combination of probes tot chromosome 1, 2 & 4 were used separately. A significant deviation of the frequency of symmetrical translocation and dicentrics from a DNA-proportional distribution was observed. Chromosomes 2, 7, 8, 9 and 21 were less frequently involved in the formation of symmetrical translocations and dicentrics than expected, whereas chromosomes 1 and 4 were more frequently involved. Chromosome 2 and 4 showed a higher frequency of acentric fragments. When triple combination probe for chromosome 1, 2 & 4 was used, no differences were found between the observed and expected frequency of exchange type aberrations. The results showed that the frequency of radiation-induced chromosome aberrations was not proportional to DNA contents, suggesting the difference in the susceptibility to specific aberrations among individual chromosomes. The results also indicated that the FISH technique with combination of probes for chromosome 1, 2 & 4 was useful for radiation biodosimetry.

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