• Korean Journal of Microbiology
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• v.43 no.1
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• pp.14-18
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• 2007

The bacterial community structures at 2 sponge species belonging to the genus Baikalospongia and Lubomirskia in Lake Baikal were analyzed with fluorescent in situ hybridization (FISH) method. The total bacterial numbers in the genus Baikalospongia ranged from $7.2{\times}10^{7}\;to\;4.2{\times}10^{8}\;cells/ml$, and those in Lubomirskia from $1.2{\times}10^{8}\;to\;1.6{\times}10^{8}\;cells/ml$, while those of lake water were from $2.3{\times}7.7{\times}10^{5}\;cells/ml$. Total bacterial numbers in the sponges were $10^{3}-10^{4}$ times higher than those of lake water. In the genera Baikalospongia and Lubomirskia, the proportions of other unidentified bacterial groups to the Bacteria were 42.0-60.3% and 40.7-51.9%, respectively. These proportions were similar to those in lake water (22.6-46.3%). These results suggest that bacterial compositions in Lake Baikal water and sponges are highly unique.

• Korean Journal of Microbiology
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• v.38 no.4
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• pp.287-292
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• 2002

The composition of bacterial communities was detected in surface water of Cheonho Reservoir dominated by cyanobacteria, using fluorescent in situ hybridization (FISH) method. Total bacterial numbers were very high ranging from 0.6～$1.3{\times}10^7 \cells{\cdot}ml^-1$, whereas the ratio of Eubacteria to total bacteria was 29.8~45.8%, which was lower than that in other freshwater ecosystems. On average only 2.1% of DAPI-stained bacteria were detected by FISH with probes for $\alpha$, $\beta$, and $\gamma$-groups, respectively. Unknown eubacteria which was not bound to any probes except EUB 338, was relatively high. On the other hand, the Cytophaga-Flavobacterium group increased following the change of dominant species from Anabaena sp. to Microcystis sp. This result showed that bacterial communities could be affected by phytoplanktons, especially cyanobacteria.

• Korean Journal of Plant Taxonomy
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• v.41 no.4
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• pp.324-328
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• 2011

The purpose of this study was to analyze the interspecific relationships of Chenopodium album and its related taxa collected in Korea. The 18S-26S ribosomal DNA (45S rDNA) loci were detected directly on mitotic chromosomes by fluorescence in situ hybridization (FISH) and the chromosome numbers were examined using aceto-orcein methods. The chromosomal numbers of Chenopodium album var. album and C. album var. centrorubrum were 2n = 6x = 54, whereas for C. album var. stenophyllum, this number was 2n = 4x = 36. The basic chromosome number was x = 9. The biotin labeled 18S-26S rDNA probe exhibited eight yellow fluorescent signals on the metaphase chromosome of C. album var. album and var. centrorubrum respectively, while two yellow signals of C. album var. stenophyllum were noted. All of the signals on the chromosomes were located at the terminal regions. The chromosome number and FISH findings suggest that C. album var. centrorubrum is merged into var. album and that it is clearly distinguished from C. album var. stenophyllum.

• 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.

• Journal of Embryo Transfer
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• v.14 no.1
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• pp.3-5
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• 1999

이식 전 수정란의 성판정은 많은 축산인의 소망이었다. 많은 방법이 제기되었지만, 세포유전학적 분석방법은 ET의 상용화에 거의 무의미할 정도로 한계가 많았고, 이후 개발된 응성 특이적 항원이나 X-염색체에서 발현되는 효소의 활동을 통한 성판정 방법은 흥미롭기는 하나 산업적 적용에 제약이 많았다. 80년대 중반 Y-염색체 특이적인 DNA 탐침자의 개발과 PCR을 통한 DNA증폭기술의 개발은 수정란 성판정을 획기적으로 개선시켰다. DNA기술을 통한 수정란의 성판정은 분자생물학을 현장으로 진출시킨 첫 경우이기도 했다. 90년대에 세포유전학적 분석에서 FISH가 소개되었으며, 염색체 특이적 library는 다른 기초적이고 응용세포유전학적 연구에 유용한 도구가 되었다. 최근에는 사람에서는 착상전 수정란의 성판별 및 유전진단을 위해 실시되고 있는 형광직접접합법(fluorescent in situ hybridization, FISH)은 효소적 유전자 증폭(polymerase chain reaction; PCR)에 비해 높은 민감도와 정확성을 보이고 있으나 hybridization 및 washing 과정에 매우 긴 시간이 소요되고, 절차도 까다로워 현장에서의 적용이 용이치 않았다. 그러나 direct labelled probe의 이용, heat programmable instrument의 개발, denaturing chemical의 사용배제 등을 통해 소요시간 및 절차의 대폭적인 간소화를 이루어 현장의 적용 가능성을 한층 높이고 있다. 현재 사람의 세포유전학 및 종양학에서는 FISH의 다양한 기술이 많이 이용하고 있으나 소에서는 탐침자(probe)가 개발되어 있지 않아 그 이용이 미미하다. 본 연구는 FISH를 이용하기 위한 탐침자의 개발을 궁극적인 목표로 삼았으며, 이를 위해 접근이 용이한 방식을 개발하여 기존의 방식과는 다른 소 배아세포의 성을 판별 할 수 있는 접근방법을 소개 하고자한다.

• Journal of Environmental Science International
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• v.21 no.12
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• pp.1455-1462
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• 2012

The vertical distributions of nitrifying bacteria in aerobic fixed biofilm were investigated to evaluate the relationship between nitrification performance and microbial community at different HRT. Fluorescent in situ hybridization (FISH) and portable ion selective microelectrode system were adopted to analyze microbial communities and ions profiles according to the biofilm depth. Cilia media packed MLE (Modified Ludzack-Ettinger) like reactor composed of anoxic, aerobic I/II was operated with synthetic wastewater having COD 200 mg/L and $NH_4{^+}$-N mg/L at HRT of 6 hrs and 4 hrs. Total biofilm thickness of aerobic I, II reactor at 4 hrs condition was over two times than that of 6 hrs condition due to the sufficient substrate supply at 4 hrs condition (6 hrs; aerobic I 380 ${\mu}m$ and II 400 ${\mu}m$, 4 hrs; aerobic I 830 ${\mu}m$ and II 1040 ${\mu}m$). As deepen the biofilm detection point, the ratio of ammonia oxidizing bacteria (AOB) was decreased while the ratio of nitrite oxidizing bacteria (NOB) was maintained similar distribution at both HRT condition. The ratio of AOB was higher at 4 hrs than 6 hrs condition and $NH_4{^+}$-N removal efficiency was also higher at 4 hrs with 89.2% than 65.4% of 6 hrs. However, the ratio of NOB was decreased when HRT was reduced from 6 hrs to 4 hrs and $NO_2{^-}$-N accumulation was observed at 4 hrs condition. Therefore, it is considered that insufficient HRT condition could supply sufficient substrate and enrichment of AOB in all depth of fixed biofilm but cause decrease of NOB and nitrite accumulation.

• Korean Journal of Animal Reproduction
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• v.20 no.3
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• pp.323-331
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• 1996

Predetermination of sex in mammalian species has many aspects of application including the prenatal diagnoses of genetic disorders in humans and sex-selected breeding programs in the animal industry. Embryos sexing can be carried out using the polymerase chain reaction (PCR) to amplify specific sequences present in the sex chromosomes, or by fluorescent in situ hybridization (FISH) of specific probes to the X and Y chromosomes. A 3.3 kb porcine male-specific DNA fragment (pEM39) was cloned previously in our laboratory. In this study, FISH and PCR methods were employed to examine if the pEM39 can be used a sex-specific DNA probes Porcine ovaries were obtained from a local slaughter house and oocytes collected. All oocytes were subjected to in vitro maturation followed by 1n vitro fertilization. Parthenogenetically activated embryos were served as a negative control. Embryonic samples were collected at the 2-cell stages and PCR was performed to analyze DNA. Among 10 embryos examined, four embryos were identified as males and six were females. The cloned male-specific DNA fragment showed male-specificity for the cells in the liver tissue and the porcine early embryos by FISH. It was also demonstrated that the cloned male-specific DNA is localized on the hetero chromatic region of the long arm in the Y chrom-osome (Yq) as shown by the FISH and karyotyping. The results suggest that the cloned male-specific DNA fragment may be useful for predetermination of sex with a few embryonic cells. The porcine male-specific sequence can be a reliable index for embryo sexing by PCR.

• Korean Journal of Ecology and Environment
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• v.37 no.4 s.109
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• pp.363-367
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• 2004

For comparative analysis of the eubacterial community structure at 8 sampling sites throughout the Nak-Dong River, FISH (fluorescence in situ hybridization) method was employed. The total ratio of each determined eubacterial group such as ${\alpha}\;{\cdot}\;{\beta}\;{\cdot}\;{\gamma}-subclasses$proteobacteria and Cytophaga-Flavobacterium(CF) group to total counts(DAPI) at each site varied 9.3-42.5% with the highest value at uppermost part. And each ratio of determined eubacterial groups reached mostly under 10% except that of CF group (23%) at uppermost part. Furthermore, compared to lower part, upper part represented unexpectedly higher proportions of ${\gamma}-subclass$ proteobacteria comprised almost fast growing bacteria on degradable organics. Also the variations of ammonia-oxidizing bacteria ranged from $2.7{\times}10^4$ to $18.0{\times}10^4$ cells $mL^{-1}$ with the lowest value in lower part and the highest value in mid part whereas those of nitrite-oxidizing bacteria varied 5.2-7.7{\times}10^4$ cells $mL^{-1}$ without noticeable differences throughout the sites. Additionally, the ratio of nitrifying bacteria to total counts ranged from 1.0% to 13.6% with no differences between ammonia-oxidizing bacteria and nitrite-oxidizing bacteria. In conclusion, FISH method introduced in this study for monitoring, normally used for the quantitative analysis of bacteria, provided also good information on their environmental status in the Nak-Dong River.

• Korean Journal of Medicinal Crop Science
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• v.26 no.2
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• pp.113-118
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• 2018

Background: Gastrodia elata Blume is a saprophytic perennial plant in the Orchidaceae family, because of its agricultural and medicinal effectiveness, researchers focus on its genome and chemical components. However, cytogenetic information based on the chromosome structure and composition to construct chromosomal backbone for genome sequencing research and for the development and breeding of plants is very limited. Methods and Results: We determined the metaphase chromosome composition of the G. elata genome by fluorescence in situ hybridization (FISH) using 5S and 45S rDNAs and telomeric repeat probes. The nuclear genome of G. elata was organized into 2 n = 36, with relatively small ($2.71-5.50{\mu}m$)chromosomes that showed gradual decrease in size. Conglutination phenomenon was observed among the metaphase chromosomes, and it was distinguished from that in other plant metaphase chromosome spreads. One pair of signal was detected for each 5S and 45S rDNA in the pericentromeric region and interstitial region on the short arm of chromosomes 10 and 4, respectively, and telomeric DNA signals were detected in the terminal region of most chromosomes. Conclusions: To our knowledge, this is the first FISH chromosome composition result in G. elata and could be useful in more comprehensive molecular cytogenetic and genomic analyses as well as breeding programs of the medicinal plant G. elata.

• Journal of Microbiology
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• v.42 no.4
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• pp.278-284
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• 2004

Changes in the soil bacterial community of a coniferous forest were analyzed to assess microbial responses to wildfire. Soil samples were collected from three different depths in lightly and severely burned areas, as well as a nearby unburned control area. Direct bacterial counts ranged from $3.3­22.6\times10^8\;cells/(g{\cdot}soil).$ In surface soil, direct bacterial counts of unburned soil exhibited a great degree of fluctuation. Those in lightly burned soil changed less, but no significant variation was observed in the severely burned soil. The fluctuations of direct bacterial count were less in the middle and deep soil lay­ers. The structure of the bacterial community was analyzed via the fluorescent in situ hybridization method. The number of bacteria detected with the eubacteria-targeted probe out of the direct bacterial count varied from $30.3\;to\;84.7\%,$ and these ratios were generally higher in the burned soils than in the unburned control soils. In the surface unburned soil, the ratios of $\alpha,\;\beta\;and\;gamma-proteobacteria,$ Cytoph­aga-Flavobacterium group, and other eubacteria groups to total eubacteria were 9.9, 10.6, 15.5, 9.0, and $55.0\%,$ respectively, and these ratios were relatively stable. The ratios of $\alpha,\;\beta\;and\;gamma-proteobacteria,$ and Cytophaga-Flavobacterium group to total eubacteria increased immediately after the wildfire, and the other eubacterial proportions decreased in the surface and middle layer soils. By way of contrast, the composition of the 5 groups of eubacteria in the subsurface soil exhibited no significant fluctuations dur­ing the entire period. The total bacterial population and bacterial community structure disturbed by wildfire soon began to recover, and original levels seemed to be restored 3 months after the wildfire.