• Title/Summary/Keyword: Fluorescence In Situ Hybridization

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

  • 김수현;김우철;김현희;허권
    • 한국약용작물학회지
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    • 제28권5호
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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.

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

  • 정영재;김무열;이병순
    • 식물분류학회지
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    • 제41권4호
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    • pp.324-328
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    • 2011
  • 한국산 흰명아주(Chenopodium album var. album), 명아주(var. centrorubrum), 가는명아주(var. stenophyllum)를 대상으로 aceto-orcein에 의한 염색체 수와 모양을 조사하였고, 45S rDNA 유전자를 이용한 FISH (fluorescence in situ hybridization) 방법을 수행하여 세포유전학적 유연관계를 고찰하였다. 체세포 염색체 수는 흰명아주와 명아주는 모두 2n = 6x = 54개인 반면에 가는명아주는 2n = 4x = 36으로 뚜렷이 구별되었으며, 기본염색체수는 x = 9 개였다. 명아주속의 염색체에서 45S rDNA의 위치를 알아보기 위한 FISH 결과는 흰명아주의 경우 8개의 signal이, 가는명아주에서는 2개의 signal이 관찰되어 종간 차이를 보였으며, 모두 염색체 말단부위에서 관찰되었다. 염색체의 수와 형태, 45S rDNA를 이용한 FISH 결과는 명아주가 흰명아주에 통합되지만, 가는명아주와는 뚜렷이 구별됨을 지지해 주었다.

Molecular Characterization of the Soybean L-Asparaginase Gene Induced by Low Temperature Stress

  • Cho, Chang-Woo;Lee, Hye-Jeong;Chung, Eunsook;Kim, Kyoung Mi;Heo, Jee Eun;Kim, Jung-In;Chung, Jongil;Ma, Youzhi;Fukui, Kiichi;Lee, Dae-Won;Kim, Doh-Hoon;Chung, Young-Soo;Lee, Jai-Heon
    • Molecules and Cells
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    • 제23권3호
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    • pp.280-286
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    • 2007
  • L-asparaginase (EC 3.5.1.1) catalyzes the hydrolysis of the amide group of L-asparagine, releasing aspartate and $NH_4{^+}$. We isolated a low temperature-inducible cDNA sequence encoding L-asparaginase from soybean leaves. The full-length L-asparaginase cDNA, designated GmASP1, contains an open reading frame of 1,258 bp coding for a protein of 326 amino acids. Genomic DNA blotting and fluorescence in situ hybridization showed that the soybean genome has two copies of GmASP1. GmASP1 mRNA was induced by low temperature, ABA and NaCl, but not by heat shock or drought stress. E. coli cells expressing recombinant GmASP1 had 3-fold increased L-asparaginase activity. A possible function of L-asparaginase in the early response to low temperature stress is discussed.

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

  • 류란숙;이상찬;이채식;김종흥;류은경;손시환
    • Clinical and Experimental Reproductive Medicine
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    • 제27권4호
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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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Smith-Magenis 증후군 2예 (Two cases of Smith-Magenis syndrome)

  • 정성관;박규희;신혜경;은소희;은백린;유기환;홍영숙;이주원;배숙영
    • Clinical and Experimental Pediatrics
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    • 제52권6호
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    • pp.701-704
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    • 2009
  • Smith-Magenis 증후군(SMS)은 17번 염색체에서 유전물질을 향유한 곳이 일부 떨어져 나가면서 생기는 질환으로, 신체, 발달 및 행동상의 특징적 이상이 나타나는 질환이다. 출생빈도는 출생아 25,000명 중에 한 명 꼴로 출생하는 것으로 알려져 있으나 최근 분자유전학적 진단 기술의 발달로 이 질환의 환자수가 점차 증가되고 있다. 다양한 임상증상과 더불어 수면장애, 경련에 대한 치료뿐만 아니라 적절한 언어, 행동학적 치료가 필요하다. 저자들은 SMS 환아 2예를 진단하고 치료하고 있는 경험이 있어 이를 보고하는 바이다.

Cytogenetic Mapping of Carthamus tinctorius L. with Tandemly Repeated DNA Sequences by Fluorescence in situ Hybridization

  • Mancia, Franklin Hinosa;Ju, Yoon Ha;Lim, Ki-Byung;Kim, Jung Sun;Nam, Sang Yong;Hwang, Yoon-Jung
    • 한국자원식물학회지
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    • 제30권6호
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    • pp.654-661
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    • 2017
  • Dual-color fluorescence in situ hybridization karyotype analysis was created using repetitive sequences including two types of rDNA repeats (45S and 5S rDNAs) and Arabidopsis-type telomere sequence repeats. The somatic metaphase cells of Carthamus tinctorius were observed as diploids (2n=2x=24). A symmetrical or slightly asymmetrical karyotype with seven pairs of metacentric and five pairs of submetacentric chromosomes was observed. The lengths of the somatic metaphase chromosomes ranged from 4.18 to $6.53{\mu}m$, with a total length of $60.71{\mu}m$. One locus of 45S rDNA was located on the pericentromeric regions of three pairs of chromosomes and the other pair was situated on the terminal regions of the short arms of a single pair of chromosomes. One locus of 5S rDNA was detected on the interstitial regions of the short arms of two pairs of chromosomes. Arabidopsis-type telomeric repeats were detected on the terminal regions of all pairs of chromosomes. Co-localization of loci between telomeric repeats and 45S rDNA was observed in a single pair of chromosomes. The results provide additional information for the existing physical mapping project of C. tinctorius and will also serve as a benchmark to a more intricate cytogenetic investigation of C. tinctorius and its related species.

소양호에서 세균군집구조의 계절적.수직적 변화 (Seasonal and Vertical Change of Bacterial Communities in Lake Soyang)

  • 김동주;홍선희;안태석
    • 미생물학회지
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    • 제35권3호
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    • pp.242-247
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    • 1999
  • 소양호에서 fluorescent in situ hybridization 방법을 이용하여 세균군집의 계절과 수심에 따른 변화를 조사하였다. 이 방법에 사용된 oligonucleotide probe는 EUB338, ALF1b, BET42a, GAM42a 와 CF probe 였다. 조사기간중 Proteobacteria $\alpha$-group 은 0.7~32.9% 이었으며 $\beta$-group은 1.0~25.8%, $\gamma$-group은 2.4~37.1%, Cytophaga-Flavobacterium group은 4.7~23.6%의 분포를 나타내었다. 계절별로 살펴보면 규조류가 우점하는 봄펄에는 $\gamma$-group 이, 유기물의 농도가 낮고 수온이 높은 여름철에는 $\alpha$-group 이 우점하였으며, 남조류가 우점하는 가을철에 총세균수에 대한 Eubacterial group 의 비율이 크게 감소하여 특정 group의 우점현상은 나타나지 않았다. 이처럼 소양호는 계절과 수심에 따라 군집구조가 변화하였으며, 특히 세균의 군집구조는 식물플랑크톤의 천이와 밀접한 관계를 보이는 것으로 나타났다.

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Fusion Between TMPRSS2 and ETS Family Members (ERG, ETV1, ETV4) in Prostate Cancers from Northern China

  • Wang, Jian-Jiang;Liu, Yue-Xin;Wang, Wei;Yan, Wei;Zheng, Yu-Peng;Qiao, Lu-Dong;Liu, Dan;Chen, Shan
    • Asian Pacific Journal of Cancer Prevention
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    • 제13권10호
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    • pp.4935-4938
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    • 2012
  • In this study we evaluated the frequency of fusion between TMPRSS2 and ETS family members (ERG, ETV1, ETV4) in prostate cancers in patients from northern China in order to explore differences in fusion rates among regions in northern and southern China, other parts of Asia, Europe, and North America. We examined 100 prostate cancer patients, diagnosed by means of prostate biopsy; fluorescence in situ hybridization (FISH) was used to detect the expression of TMPRSS2, ERG, ETV1 and ETV4 in cancer tissue. Differences in gene fusion rates among different ethnics groups were also analyzed. Of the 100 prostate cancer patients, 55 (55%) had the fusion gene. Among the patients with the fusion gene, 46 (83.6%) patients had the TMPRSS2:ERG fusion product, 8 (14.8%) patients had TMPRSS2:ETV1 fusion, 1 (1.6%) patient had TMPRSS2:ETV4.

rDNA와 말단소체 반복서열 탐침을 이용한 천마의 FISH 염색체 조성 분석 (Analysis of Chromosome Composition of Gastrodia elata Blume by Fluorescent in situ Hybridization using rDNA and Telomeric Repeat Probes)

  • ;박응준;김현희
    • 한국약용작물학회지
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    • 제26권2호
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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.

Circulating Aneuploid Cells Detected in the Blood of Patients with Infectious Lung Diseases

  • Kim, Hongsun;Cho, Jong Ho;Sonn, Chung-Hee;Kim, Jae-Won;Choi, Yul;Lee, Jinseon;Kim, Jhingook
    • Journal of Chest Surgery
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    • 제50권2호
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    • pp.126-129
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    • 2017
  • The identification of circulating tumor cells (CTCs) is clinically important for diagnosing cancer. We have previously developed a size-based filtration platform followed by epithelial cell adhesion molecule immunofluorescence staining for detecting CTCs. To characterize CTCs independently of cell surface protein expression, we incorporated a chromosomal fluorescence in situ hybridization (FISH) assay to detect abnormal copy numbers of chromosomes in cells collected from peripheral blood samples by the size-based filtration platform. Aneuploid cells were detected in the peripheral blood of patients with lung cancer. Unexpectedly, aneuploid cells were also detected in the control group, which consisted of peripheral blood samples from patients with benign lung diseases, such as empyema necessitatis and non-tuberculous mycobacterial lung disease. These findings suggest that chromosomal abnormalities are observed not only in tumor cells, but also in benign infectious diseases. Thus, our findings present new considerations and bring into light the possibility of false positives when using FISH for cancer diagnosis.