• Title/Summary/Keyword: Fluorescence in-situ hybridization (FISH)

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Cytotaxonomical Study of the Chenopodium album and its Related Species in Korea (한국산 흰명아주와 근연종의 세포분류학적 연구)

  • Chung, Youngjae;l Kim, Muyeol;Lee, Byongsoon
    • 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.

Identification of a Human Y Chromosome Specific DNA Probe and Their Clinical Application by Fluorescence in situ Hybridization Techniques (사람 Y 염색체 특이 DNA Probe의 개발과 이를 이용한 FISH 기술의 임상적 적용)

  • Ryu, Ran-Suk;Lee, Sang-Chan;Lee, Chae-Sik;Kim, Jong-Heung;Ryu, Eun-Koung;Sohn, Sea-Hwan
    • Clinical and Experimental Reproductive Medicine
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    • v.27 no.4
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    • pp.405-415
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    • 2000
  • Objective: The present study was undertaken to synthesize a human Y chromosome specific probe and to confirm the usefulness of the probe for fluorescence in situ hybridization (FISH) in various types of human cells. Methods: An approximately 400 bp DNA fragment of the DYZ1 sequences was synthesized by PCR using digoxigenin labeled dUTP (dig-PCR). The fidelity of probe was tested by FISH for cultured and uncultured human lymphocytes, amniocytes, chorionic villus cells, embryos, sperms, and germ cells of seminiferous tubule. Results: The human Y chromosome specific probe hybridized specifically to Y chromosome of the cells that had been tested. This probe assigned to the Yq12 region where the DYZ1 repetitive sequence is concentrated. Conclusion: We have identified a human Y chromosome specific probe that hybridized specifically to the Y chromosome by FISH for various types of uncultured as well as cultured cells. Therefore FISH technique using human Y chromosome specific probe should be useful for clinical application as a diagnostic tool for the detection of human Y chromosome.

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Multi-dimensional analyses of plant chromosomes and genomes.

  • Fukui, Kiichi;Ohmido, Nobuko;Wako, Toshiyuki
    • Proceedings of the Botanical Society of Korea Conference
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    • 1998.07a
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    • pp.61-70
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    • 1998
  • Genome and chromosome analyses in plants using fluorescence in situ hybridization (FISH) and immuno-staining (IMS) methods are reviewed by presenting the recent results obtained by the Chromosome Link, a group of chromosome and genome researchers. FISH is now effective to detect unique nucleotide sequences with 153 bp on the extended DNA fibers. Genomic in situ hybridization (GISH) also allows painting plant chromosomes of different genomes. GISH is quite effective to detect the genomic differentiation in the individual chromosomes within a nucleus. Three dimensional (3D) analyses are now available by confocal microscopy and a deconvolution system. These techniques are invaluable to visualize both the structural and functional dynamics within a nucleus. 3D-FISH revealed the spatial differentiation of different genomees within a nucleus. 3D-FISH also proved structural partition of centromeric and telomeric domains within a barely nucleus. The dynamic acetylation of histone H4 at the specific regions of a genome during a cell cycle is also analyzed using 3D-IMS. It is anticipated that these methods will provide us powerful tools to understand the structural and functional significance of plant chromosomes and genomes.

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Diagnostic Value of Fluorescence in Situ Hybridization Assay in Malignant Mesothelioma: A Meta-analysis

  • Wan, Chun;Shen, Yong-Chun;Liu, Meng-Qi;Yang, Ting;Wang, Tao;Chen, Lei;Yi, Qun;Wen, Fu-Qiang
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.9
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    • pp.4745-4749
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    • 2012
  • The diagnosis of malignant mesothelioma (MM) remains a clinical challenge and the fluorescence in situ hybridization (FISH) assay has been reported to be one promising tool. The present meta-analysis aimed to establish the overall diagnostic accuracy of FISH for diagnosing MM. After a systematic review of English language studies, the sensitivity, specificity and other measures of accuracy of FISH in the diagnosis of MM were pooled using random-effects models. Summary receiver operating characteristic curves were applied to summarize overall test performance. Nine studies met our inclusion criteria, the pooled sensitivity and specificity for FISH for diagnosing MM being 0.72 (95% CI 0.67-0.76) and 1.00 (95% CI 0.98-1.00), respectively. The positive likelihood ratio was 34.5 (95% CI 14.5-82.10), the negative likelihood ratio was 0.24 (95% CI 0.16-0.36), and the diagnostic odds ratio was 204.9 (95% CI 76.8-546.6), the area under the curve being 0.99. Our data suggest that the FISH assay is likely to be a useful diagnostic tool for confirming MM. However, considering the limited studies and patients included, further large scale studies are needed to confirm these findings.

Dynamics of in situ Bacterial Community Structure in the Nak-Dong River (낙동강에서의 세균군집구조의 역동성)

  • Park, Ji-Eun;Yeo, Sang-Min;Lee, Young-Ok
    • 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.

Wastewater Treatment and Microbial Structure Analysis by Fluorescence In Situ Hydridizationin a Biofilm Reactor (생물막 반응키에서의 폐수 처리 및 Fluorescence In Situ Hybridization에 의한 복합 미생물계 구조 해석)

  • Kim, Dong-Jin;Han, Dong-Woo;Lee, Soo-Choul;Park, Byeong-Gon;Kwon, Il;Sung, Chang-Keun;Park, Wan-Cheol
    • KSBB Journal
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    • v.17 no.1
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    • pp.80-87
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    • 2002
  • Laboratory scale aerobicfanaerobic biofilm reactor was used for simultaneous and stable removal of organics, N and P components to investigate optimum design and operation parameters and to analyze the microbial distribution and consortium structure of nitrification and denitrification bacteria in aerobic and anaerobic biofilm systems. The biofilm reactor was successfully operated for 143 days to show $COD_{cr},\;BOD_5$, SS removal efficiencies of 88, 88, and 97%, respectively. During the experiment period, almost complete nitrification efficiency of 96% was achieved. Denitrification efficiency was about 45% without addition of any external carbon sources. In case of total phosphorus removal, 74% of the inlet phosphorus was removed. Fluorescence in situ hybridization (FISH) results showed that most of the ammonia oxidizing bacteria in the aerobic nitrification zone was found to be Nitrosomonas species while Nitrospira was the representative nitrite oxidizing bacteria. For the denitrification, Rhodobacter, Rhodovulum, Roseebacter and Paracouus were the dominant denitrification bacteria which was 10 to 20% of the total bacteria in numbers.

The Effect of Fixed Media and Recycling Ratio on Nutrients Removal in a Pilot-Scale Wastewater Treatment Unit (고정식 담체 유무와 반송비에 따른 소규모 하수처리 시스템 내 영양염류 제거 특성)

  • Hwang, Jae-Hoon;Cho, Dong-Wan;Kim, Chung-Hwan;Jeon, Byong-Hun
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.6
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    • pp.449-455
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    • 2013
  • The effect of recycling ratio and fixed media on nitrate and phosphate removal was investigated in a pilot-scale wastewater treatment unit using synthetic wastewater. Addition of fixed media increased nitrate removal from 45 to 58% while no noticeable change was observed for Chemical Oxygen Demand (COD) and phosphate removal (<5%). Nitrate removal efficiency also enhanced (Ca 7%) when the influent wastewater flow was doubled (2Q), however phosphate removal was decreased from 40.9 to 26.6% with the increasing recycling rate. The attached biomass analysis showed the presence of bacteria (73.4 $mg/cm^2$) on the surface of added media in anoxic reactor. Pseudomonas aeruginosa a common denitrifying bacterium dominated the bacterial growth (58%) in the anoxic reactor which was determined using Fluorescence In Situ Hybridization (FISH) analysis.

The first Korean case of a newborn with 3p26 microdeletion and 5q35 microduplication inherited from paternal balanced translocation

  • Jang, Jin A;Sohn, Young Bae;Lee, Jang Hoon;Park, Moon Sung
    • Journal of Genetic Medicine
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    • v.18 no.1
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    • pp.48-54
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    • 2021
  • Genetic imbalances are a major cause of congenital and developmental abnormalities. We report the first case of a 3p26 microdeletion and 5q35.2q35.3 microduplication in a newborn with multiple congenital anomalies evaluated using chromosomal microarray analysis (CMA) and fluorescence in situ hybridization (FISH). The patient was born at 30 weeks and 2 days of gestation with a body weight of 890 g. He had symmetric intrauterine growth restriction, microcephaly, facial dysmorphism (hypertelorism, blepharophimosis, mild low-set ears, high-arched palate, and micrognathia), and right thumb polydactyly. Echocardiography revealed an atrial septal defect and patent ductus arteriosus. Furthermore, CMA revealed a concurrent microdeletion in 3p26 and a microduplication in 5q35.2q35.3. FISH analysis showed that these genetic changes resulted from a translocation mutation between chromosomes 3 and 5. The patient's mother had mild intellectual disability, short stature, and facial dysmorphism, while his father had a normal phenotype. However, parental FISH analysis revealed that the asymptomatic father carried a balanced translocation of chromosomes 3p26 and 5q35. CMA and FISH tests are useful for diagnosing neonates with multiple congenital abnormalities. Further parental genetic investigation and proper genetic counseling are necessary in cases of chromosomal abnormalities inherited from parental balanced translocations.

Karyotype Analysis and Physical Mapping of rDNAs in Bupleurum longeradiatum (개시호 (Bupleurum longeradiatum)의 핵형분석과 rDNAs의 Physical Mapping)

  • Koo, Dal-Hoe;Seong, Nak-Sul;Seong, Jong-Suk;Bang, Kyong-Hwan;Bang, Jae-Wook
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.5
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    • pp.402-407
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    • 2003
  • Karyotype analysis and chromosomal localization of 5S and 45S rDNAs using multi-color fluorescence in situ hybridization (McFISH) technique were carried out in Bupleurum longeradiatum. Somatic metaphase chromosome number was 2n=12. Karyotype was composed of three pairs of metacentrics (No.3, 4 and 6) and three pairs of submetacentrics (No. 1, 2 and 5). The length of somatic prometaphase chromosomes ranges from 2.55 to $5.05{\mu}m$ with total length of $18.15\;{\mu}m$. In FISH experiment, one pair of 5S rDNA signals was detected on the pericentromeric region of chromosome 4 and one pair of 45S rDNA signals was detected on the telomeric region of chromosome 2.

Cytogenetical Study of Cynanchum wilfordii and Cynanchum auriculatum using Fluorescence In Situ Hybridization (FISH) (rDNA FISH를 이용한 큰조롱과 넓은잎큰조롱의 세포유전학적 연구)

  • Kim, Su Hyun;Kim, Woo Cheol;Kim, Hyun Hee;Heo, Kweon
    • Korean Journal of Medicinal Crop Science
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    • v.28 no.5
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    • pp.325-330
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    • 2020
  • Background: Dried tuberous roots of Cynanchum wilfordii are known to relieve menopause symptoms. However, the dried roots of C. wilfordii are morphologically similar to those of C. auriculatum, which makes it difficult to distinguish when used as a medicine. Various comparative studies have focused on chemical or molecular analysis of these roots. However, the differences between the two species at the cytogenetic level based on chromosome structure and composition remain to be elucidated. Methods and Results: For chromosome slides, the roots were fixed in 8-hydroxyquinoline, digested with enzyme mixture, and spread on slides. 5S and 45S rDNA were used as cytogenetic markers for the analysis of nuclear genomes by FISH. The chromosome number of the two species was 2n = 22, with a relatively short length, 1.13 ㎛ - 4.24 ㎛ and 1.00 ㎛ - 3.42 ㎛ with respect to each other. Both species represent one pair of 5S and 45S rDNA signal on chromosome 1, at the proximal region and peri-centromeric region, respectively. Conclusions: These preliminary cytogenetic data using FISH in C. wilfordii and C. auriculatum could be valuable for the comprehension of Cynanchum genome history.