• Title/Summary/Keyword: Fluorescent probes

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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.

Molecular Identification of Korean Ginseng Cultivars (Panax ginseng C. A. Mey.) using Peptide Nucleic Acid (PNA) Microarray (PNA 바이오칩을 이용한 고려인삼 품종 판별)

  • Bang, Kyong-Hwan;Jo, Ick-Hyun;Kim, Young-Chang;Kim, Jang-Uk;Park, Hong-Woo;Shin, Mi-Ran;Kim, Young-Bae;Kim, Ok-Tae;Hyun, Dong-Yun;Kim, Dong-Hwi;Cha, Seon-Woo
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.5
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    • pp.387-392
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    • 2012
  • This study was carried out to identify Korean ginseng cultivars using peptide nucleic acid (PNA) microarray. Sixty-seven probes were designed based on nucleotide variation to distinguish Korean ginseng cultivars of Panax ginseng. Among those PNA probes, three (PGB74, PGB110 and PGB130) have been developed to distinguish five Korean ginseng cultivars. Five Korean ginseng cultivars were denoted as barcode numbers depending on their fluorescent signal patterns of each cultivar using three probe sets in the PNA microarray. Five Korean ginseng cultivars, Chunpoong, Yunpoong, Gopoong, Gumpoong and Sunpoong, were simply denoted as '111', '222', '211', '221' and '122', respectively. This is the first report of PNA microarray which provided an objective and reliable method for the authentication of Korean ginseng cultivars. Also, the PNA microarray will be useful for management system and pure guarantee in ginseng seed.

Effects of Local Anesthetics on Rotational Mobility of n-(9-Anthroyloxy)stearic Acid in Neuronal Membranes

  • Jang, Hye-Ock;Lee, Chang;Choi, Min-Gak;Shin, Sang-Hun;Chung, In-Kyo;Yun, Il
    • The Korean Journal of Physiology and Pharmacology
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    • v.7 no.3
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    • pp.119-124
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    • 2003
  • To elucidate the molecular mechanism of pharmacological action of local anesthetics, we studied membrane actions of tetracaine, bupivacaine, lidocaine, prilocaine and procaine. Fluorescence polarization of n-(9-anthroyloxy)stearic acid (n-AS) was used to examine the effects of these local anesthetics on differential rotational mobility of different positions of the number of synaptosomal plasma membrane vesicle (SPMV) phospholipid carbon atoms. The four membrane components differed with respect to 3, 6, 9 and 16-(9-anthroyloxy)stearic acid (3-AS, 6-AS, 9-AS and 16-AP) probes, indicating that differences in the membrane fluidity might be present. Degrees of the rotational mobility of 3-AS, 6-AS, 9-AS and 16-AP were different depending on depth of hydrocarbon interior. In a dose-dependentmanner, tetracaine, bupivacaine, lidocaine, prilocaine and procaine decreased anisotropy of 3-AS, 6-AS, 9-AS and 16-AP in the hydrocarbon interior of the SPMV. These results indicate that local anesthetics have significant disordering effects on hydrocarbon interior of the SPMV, thus affecting the transport of $Na^+$ and $K^+$ in nerve membranes and leading to anesthetic action.

The Bacterial Communities Structure and Its Environmental Determinants in Lake Soyang (소양호 세균군집구조와 그 구조에 영향을 주는 환경요인)

  • 김동주;홍선희;최승익;안태석
    • Korean Journal of Microbiology
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    • v.36 no.2
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    • pp.136-141
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    • 2000
  • The temporal variation of bacterial community and environmental factors, affecting on bacterial community structure were estimated monthly kom April, 1998 to May, 1999. Bacterial community structures were determined by in situ hyblidization with rRNA-targeted fluorescently labeled oligonucleotide probes (FISH) and epifluorescence microscopy; and the statistical analysis was done by SPSS program. The oligonucleotide probes used in this study were EUB338, ALFlb, GAM42a, and CF. In surface water, $\alpha$-group was related to only DOC (-0.538, p<0.05) and Chlorophyll a concentration was related to y-group (-0.630, p$\beta$-group and Cytophaga-Flavobacterium group were related to water temperature as 0.665, and 0.685 @<0.05). Between pH and $\beta$-group, there was a positive relationship (0.541, p<0.05), and Cytophaga-Flavobactevizim group was represent to correlation (0.672, p

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Seasonal Changes of bacterial community analysed by fluorescent in situ hybridization method in Lake Soyang (Fluorescent In Situ Hybridization방법으로 분석한 소양호 세균 군집 구조의 계절적 변화)

  • Hong, Sun-Hee;Ahn, Tae-Seok
    • Korean Journal of Microbiology
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    • v.34 no.3
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    • pp.169-174
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    • 1998
  • To define the structure and diversity of bacterial communities in the aqutic ecosystem, Lake Soyang, the largest artificial reservoir in Korea, a new method, fluorescent in situ hybridization was applied. This technique relies on the specific hybridization of the nucleic acid probes to the naturally amplified intracellular rRNA. By this method, the bacterial community composition of Lake Soyang and bacterial numbers belong to eubacteria, proteobacteria and Cytophaga-Flavobacterium group were estimated. Total bacterial numbers ranged from $0.3{\times}10^6{\sim}2.0{\times}10^6cells{\cdot}ml^{-1}$, and vertical profile of total bacteria showed the peak at 2 and 5 m depths. The ratio of eubacteria to total bacteria were 22~100% and varied with depth and season. The percentage of Proteobacteria ${\alpha}$-group ranged 2.6~66.7%, ${\beta}$-group 4.5~53.5%, ${\gamma}$-group 4.6~76.7% and Cytophaga-Flavobacterium group 2.1~35.9%. Also, bacteria] community had spatial and temporal characteristics. The dominant groups were ${\beta}$-group in winter, ${\gamma}$-group in spring and early summer and ${\alpha}$-group in summer.

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Succession of Bacterial Populations in Cattle Manure Compost as Determined by Fluorescent In Situ Hybridization (우분 퇴비화에서의 Fluorescent In Situ Hybridization법에 의한 세균군집의 천이)

  • Lee, Young-Ok;Jo, Ik-Hwan;Kim, Kil-Woong
    • Journal of the Korea Organic Resources Recycling Association
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    • v.8 no.2
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    • pp.146-153
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    • 2000
  • To elucidate succession of bacterial populations, especially nitrifying bacteria during the composting of cattle manure with apple pomace, fluorescent in situ hybridization(FISH) using rRNA targeted oligonucleotide probes were applied. The density of ammonia-oxidizing bacteria was ranged from $3,3{\times}10^6cells/g$ dw to $13,4{\times}10^6cells/g$ dw with the peak value after 26 composting days whereas that of nitrite-oxidizing bacteria varied between $6.0{\times}10^6cells/g$ dw and $17.2{\times}10^6cells/g$ dw with the peak value after 7 composting days. And the tendency that the numbers of nitrite-oxidizing bacteria were higher than those of ammonia-oxidizing bacteria, and the peak-time of their densities were the same as that of data determined by the ratio of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria to eubacteria. The peak of ammonia-oxidizing bacteria followed the peak of nitrite-oxidizing bacteria, at the late phase of composting process could be probably caused by the depletion of volatile ammonia of composting materials. Besides these results indicate that FISH method is a useful tool for detection of slow growing nitrifying bacteria.

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Rapid prenatal diagnosis of spinocerebellar ataxia type 3 by using fluorescent PCR

  • Kim, Do-Jin;Park, So-Yeon;Kim, Mi-Jin;Lee, Moon-Hee;Shim, Sung-Han;Ryu, Hyun-Mee
    • Journal of Genetic Medicine
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    • v.4 no.1
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    • pp.84-87
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    • 2007
  • Spinocerebellar Ataxia Type 3 (SCA 3) is a rare autosomal dominative disorder in which one of the neurodegenerative disorders is caused by a CAG repeat expansion on chromosome 14q32.1. The age at onset of disease is related to the size of the expanded CAG repeat. We present the prenatal diagnosis of SCA3 in a woman whose husband was known to carry an unstable CAG repeat expansion in the MJD gene. The diagnosis was made using PCR with a fluorescent probe for an expanded MJD allele. The normal ranges of (CAG)n of SCA3 are 14~38 repeats. The husband, who had a family history of SCA 3, has an expanded allele of 69 CAG repeats with a normal allele of 27 repeats. His wife had two normal alleles with 26 and 32 CAG repeats. The fetus had two normal alleles with 26 and 27 CAG repeats; consequently, the baby w as healthy. We report a case of prenatal diagnosis of SCA3 using a fluorescent PCR which is rapid and accurate.

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Development of FISH Methods for Detection of Genus Bacillus (Bacillus 속 세균을 검출하기 위한 Fluorescent In Situ Hybridization 방법의 개발)

  • 홍선희;김옥선;송홍규;이동훈;안태석
    • Korean Journal of Microbiology
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    • v.37 no.3
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    • pp.204-208
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    • 2001
  • A technique for detection of Bacillus in soil and waste water treatment system was developed. Mixed col- tured solutions of 22 Bacillus strains were applied for selection of probe and pretreatment method for FISHAmong the three probes known as useful tool for FISH method,S-G-Bacill-0597-a-A-22 was best for detec-tion of Bacillus. Ninety five percent of DAPl count was observed with FISH method with S-G-Bacill-0597-a-A-22 probe. For increasing the permeability to Bacillus cell walls, pretreatment with lysozyme was betterthan that with lysozyme and SDS/DTT (dithiothreitol). Bacillus spore was not detected with FISH. So, Bacil-lus detection in ecosystem requires FISH with pretreatment of lysozyme and spore staining.

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Duplication of intrachromosomal insertion segments $4q32{\rightarrow}q35$ confirmed by comparative genomic hybridization and fluorescent $in$ $situ$ hybridization

  • Kim, Jin-Woo;Park, Ju-Yeon;Oh, Ah-Rum;Choi, Eun-Young;Ryu, Hyun-Mee;Kang, Inn-Soo;Koong, Mi-Kyoung;Park, So-Yeon
    • Clinical and Experimental Reproductive Medicine
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    • v.38 no.4
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    • pp.238-241
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    • 2011
  • A 35-year-old man with infertility was referred for chromosomal analysis. In routine cytogenetic analysis, the patient was seen to have additional material of unknown origin on the terminal region of the short arm of chromosome 4. To determine the origin of the unknown material, we carried out high-resolution banding, comparative genomic hybridization (CGH), and FISH. CGH showed a gain of signal on the region of $4q32{\rightarrow}q35$. FISH using whole chromosome painting and subtelomeric region probes for chromosome 4 confirmed the aberrant chromosome as an intrachromosomal insertion duplication of $4q32{\rightarrow}q35$. Duplication often leads to some phenotypic abnormalities; however, our patient showed an almost normal phenotype except for congenital dysfunction in spermatogenesis.

Membrane-Ordering Effects of Barbiturates on Pure Phospholipid Model Membranes

  • Knag, Jung-Sook;Chung, Young-Za;Cho, Goon-Jae;Byun, Won-Tan;Yun, Il
    • Archives of Pharmacal Research
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    • v.15 no.3
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    • pp.196-203
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    • 1992
  • Intramolecular excimer formation of 1, 3-di(1-pyrenyl)propane (Py-3-Py) and fluorescence polarization of 1, 6-diphenyl-1, 3, 5-hexatriene (DPH) were used to investigate the effects of barbiturates on the fluidity of model membranes of phosphatidycholine (SPMVPC), phosphatidylserine (SPMVPS), and phosphatidylinositol (SPMVPI) fractions of synaptosomal plasma membrane vesicles (SPMV) isolated from bovine cerebral cortex. In a dose-dependent manner, barbiturates decreased the excimer to monomer fluorescence intensity ratio (I'/I) of Py-3-Py and increased the anisotropy(r), rotational relaxation time (P), limiting anisotropy $(r_infty)$, and order parameter (S) of DPH in SPMVPC, SPMVPS and SPMVPI. This indicates that barbiturates decreased both the lateral and rotational diffusion of the probes in SPMVPC, SPMVPS and SPMVPI. The relative potencies of barbiturates in ordering the membranes were in the order: pentobarbital > hexobarbital > amobarbital > phenobarbital. This order correlates well with the anesthetic potencies of barbiturates and the potencies for enhancement of $\gamma$-aminobutyric acid-stimulated chloride uptake. Thus, it is strongly suggested that a close relationship might exist between the membrane ordering effects of barbiturates and the chloride fluxes across SPMV.

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