• Title/Summary/Keyword: Fluorescence In Situ Hybridization

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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.

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.

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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    • v.23 no.3
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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.

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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Two cases of Smith-Magenis syndrome (Smith-Magenis 증후군 2예)

  • Jung, Seong Kwan;Park, Kyu Hee;Shin, Hae Kyung;Eun, So Hee;Eun, Baik-Lin;Yoo, Kee Hwan;Hong, Young Sook;Lee, Joo Won;Bae, Sook Young
    • Clinical and Experimental Pediatrics
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    • v.52 no.6
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    • pp.701-704
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    • 2009
  • SmithMagenis syndrome (SMS) is a rare disorder with multiple congenital anomalies caused by a heterozygous interstitial deletion involving chromosome 17p11.2, where the retinoic acid-induced 1 (RAI1) gene is located, or by a mutation of RAI1. Approximately 90% of the patients with SMS have a detectable 17p11.2 microdeletion on fluorescence in-situ hybridization (FISH). SMS is characterized by mental retardation, distinctive behavioral features, craniofacial and skeletal anomalies, speech and developmental delay, and sleep disturbances. Although there are some intervention strategies that help individuals with SMS, there are no reported specific interventions for improving the outcome in children with SMS. Here, we report two cases of SmithMagenis syndrome.

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
    • Korean Journal of Plant Resources
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    • v.30 no.6
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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.

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

  • 김동주;홍선희;안태석
    • Korean Journal of Microbiology
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    • v.35 no.3
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    • pp.242-247
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    • 1999
  • This sludy was conducted to investigate the change of bacterial co~munities with season and depth in Lake Soyang. Korea, using fluorescence in situ hybridization (FISH). The oligonucleotide probes used in this study were EUB338, ALF I b. GAM42a, and CF, The percentage of h e Proleobacteria a $\alpha$-group ranged from 0.70 to 33% the $\beta$-group from 1.0 to 26% they -group from 2.4 Lo 37% and Cytophagn and Flavobactefin groups from 4.7 to 24% duing the study period (April Lo November, 1998). They $\gamma$-group was dominant in spring when Asterionella was dominant. and a $\alpha$-group was dominant in summer when the organic content was low and Dinobryon was dominant. However, a specific group was not dominant in ?dl when cyanobacteria group was dominant and the ratio of eubacleria to total bacteria was very low. Therefore, the bacterial communities in Lake Soyang changed with season and depth, which seems to be associated with the telnporal succession of phytoplanlaons.

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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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    • v.13 no.10
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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.

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

  • Zhou, Hui Chao;Park, Eung Jun;Kim, Hyun Hee
    • 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.

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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    • v.50 no.2
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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.