• Title/Summary/Keyword: specific probes

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Sex Determination in Somatic and Embryonic Cells of the Pig by FISH and PCR (FISH와 PCR에 의한 돼지 체세포 및 배아세포의 성 판정)

  • Chung, Y.;Jeon, J.T.;Kim, K.D.;Lee, S.H.;Hong, K.C.
    • 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.

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Cloning of Epidermis-specific cDNAS Encoding a Lipid Transfer Protein and an Aldehyde Decarbonylase from Senecio odorus

  • Pyee, Jaeho
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.189-195
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    • 1996
  • The major cuticular components have been shown to be synthesized in the epidermis. Therefore, cloning of epidermis-specific genes could yield information to be used to isolate and characterize the enzymes involved in the cuticle biosynthesis. A subtractive cDNA library was prepared from Senecio odorus in which epidermis-specific cDNAs were enriched. Differential screening of the library using epidermal and non-epidermal probes revealed two cDNAs. One of them designated epi425 was identified, based on the sequence homology, as a member of a new class in the LTP gene family and the other clone designated epi23 as a gene encoding an aldehyde decarbonylase. Northern blot analyses showed that epi425 and epi23 cDNAs hybridized with a transcript of about 600 and 2, 100 nucleotides, respectively, from the epidermis but not from the non-epidermal tissues. Further characterization of these clones will provide more information on the mechanism of the cuticle biosynthesis.

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Using Service Design Tools in Community Nutrition Research: A Case Study in Developing Dietary Guidelines for Young Adults (서비스 디자인 도구의 지역사회영양학 분야 활용: 청년 식생활 가이드 개발 사례)

  • Jo, Eunbin;Shim, Jae Eun;Ryou, Hyun Joo;Kim, Kirang;Song, Su Jin;Kim, Hyun Ja;Ahn, Jeong Sun;Kwon, Kwang-il;Lee, Hye Young;Park, Sohyun
    • Korean Journal of Community Nutrition
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    • v.27 no.3
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    • pp.177-191
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    • 2022
  • Objectives: Recent epidemiological data reported that young adults in their 20 ~ 30s are a vulnerable population with unhealthy dietary practices and a few signs of deteriorated health indicators. However, there are no dietary guidelines that are specifically developed for the young adult population. This study introduces some data collection tools that are mostly used in the service design field, and demonstrates how these tools can be used in nutrition research for developing dietary guidelines for specific target groups. Methods: To understand the context of food choices among young people, 39 people were enrolled to complete a probes booklet. Thematic analysis and word cloud were performed to capture the main themes from the probes and a persona was developed based on the findings. Results: Data from the probes enabled us to grasp the various contextual meanings of eating practices among young people. Most participants understand what a healthy diet is and often have a willingness to practice it. However, there were very few participants who were following the practices. We created four types of persona for developing dietary guidelines: healthy eating, emotional eating, convenient eating, and trendy eating. Conclusions: Probes and persona were used in order to understand the lives of young adults and develop targeted messages. We hope that this introduction will be helpful to researchers who are looking for new ways of understanding their target population in the field of community nutrition.

Indicator-free DNA Chip Array Using an Electrochemical System

  • Park, Yong-Sung;Kwon, Young-Soo;Park, Dae-Hee
    • KIEE International Transactions on Electrophysics and Applications
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    • v.4C no.4
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    • pp.133-136
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    • 2004
  • This research aims to develop a DNA chip array without an indicator. We fabricated a microelectrode array through photolithography technology. Several DNA probes were immobilized on an electrode. Then, target DNA was hybridized and measured electrochemically. Cyclic-voltammograms (CVs) showed a difference between the DNA probe and mismatched DNA in an anodic peak. This indicator-free DNA chip resulted in a sequence-specific detection of the target DNA.

Applications of Geostatistics to the Quantitative Analysis of Genetic Instability in Carcinogenesis

  • Kim Hyoung-Moon
    • Communications for Statistical Applications and Methods
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    • v.13 no.1
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    • pp.167-175
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    • 2006
  • It has long been recognized that cancer is a genetic disease. To find this measures of genetic instability, stain cells with chromosome specific probes using chromosome in-situ hybridization technique is adopted. Even though in-situ hybridization technique is powerful, truncation of nuclei often results in under-representation of chromosome copies in slides due to the sectioning of tissue blocks. Because of this problem we suggest three different methods to analyze the cervical cancer data set. We observe that genetic instability is an increasing function of histology and our suggested model is the best in detecting genetic instability of tumorigenesis processes.

Development of Repetitive DNA Probes for Genetic Analysis of Phytophthora capsici (Phytophthora capsici의 유전적 특성 분석을 위한 Repetitive DNA Probe의 개발)

  • Song, Jeong-Young;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.66-72
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    • 2002
  • To develop DNA markers for analysis of genetic characteristics of Phytophthora capsici population, randomly selected clones from HindIII-digested genomic DNA library of P. capsici 95CY3119 were surveyed by hybridizing to Southern blots of HindIII-digested total genomic DNA of P. capsici. Probe DNAs inserted into selected individual clones strongly hybridized with HindIII digests of P. capsici. Among probes examined, PC9 revealed the repetitive and highly polymorphic bands to HindIII digests of inter-and intra-field P. capsici isolates. Genetic diversity of individual isolates was also clearly revealed in cluster analysis based on its band patterns. The other probe, PC22, was hybridized only to DNA from P. capsici and this was highly repetitive. However, there was no response to other Phytophthora species and Pythium sp. These DNA probes could be used as very useful markers in analysing genetic diversity and identification for P. capsici population throughout the world.

Microarray Probe Design with Multiobjective Evolutionary Algorithm (다중목적함수 진화 알고리즘을 이용한 마이크로어레이 프로브 디자인)

  • Lee, In-Hee;Shin, Soo-Yong;Cho, Young-Min;Yang, Kyung-Ae;Zhang, Byoung-Tak
    • Journal of KIISE:Software and Applications
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    • v.35 no.8
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    • pp.501-511
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    • 2008
  • Probe design is one of the essential tasks in successful DNA microarray experiments. The requirements for probes vary as the purpose or type of microarray experiments. In general, most previous works use the simple filtering approach with the fixed threshold value for each requirement. Here, we formulate the probe design as a multiobjective optimization problem with the two objectives and solve it using ${\epsilon}$-multiobjective evolutionary algorithm. The suggested approach was applied in designing probes for 19 types of Human Papillomavirus and 52 genes in Arabidopsis Calmodulin multigene family and successfully produced more target specific probes compared to well known probe design tools such as OligoArray and OligoWiz.

STRUCTURAL PERTURBATIONS INDUCED BY PHOTODYNAMIC ACTION OF PORPHYRIN AGGREGATES ON PLASMA MEMBRANE AND MICROSOMES OF GLIOBLASTOMA CELLS

  • Sreentvasan, Rajesh;Joshi, Preeti G.;Joshi, Nanda B.
    • Journal of Photoscience
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    • v.4 no.2
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    • pp.41-48
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    • 1997
  • The plasma membrane and microsomes, isolated from the cells treated with hematoporphyrm derivative (HpD) for 1 and 24 h, accumulated the aggregated porphyrin. The quantity of aggregated porphyrin was same in the plasma membrane and microsomes after isolating them from cells treated with HpD for 1 h whereas the microsomes accumulated higher quantity of aggregated porphyrin when cells were treated with HpD for 24 h. Photodynamic action of aggregated porphyrin on plasma membrane and microsomes was investigated using lipid specific fluorescent probes: 1,6-diphenyl-1,3,5-hexatrine (DPH) and 1-(4-trimethylammonium), 6-diphenyl-1,3,5-hexatrine(TMA-DPH). The time dependent anisotropy of these probes in the membranes was measured and the decay of anisotropy was analyzed using wobbling in cone model. Upon irradiation both the plasma membrane and the microsomes showed an increase in the limiting anis~)tropy and order parameter and a decrease in the cone angle of the lipid probes. The increase in the limiting anisotropy was pronounced in membranes isolated from the cells treated with HpD for 24 h. Photoinduced change in the limiting anisotropy was dependent on the duration of incubation of cells with HpD before isolating the membranes. In both the membranes. the membrane core was affected more as compared to the outer leaflet. In addition to the structural changes, a decrease in Na$^+$-K$^+$-ATPase and NADPH cyt c reductase activity was also observed upon irradiation of HpD treated cells. Inhibition in NADPH cyt c reductase was more when cells were treated with HpD for 24 h, however, Na$^+$-K$^+$-ATPase activity did not depend on the duration of the treatment of cells with HpD before irradiation. Our results suggest that the extent of photoinduced perturbations in the membranes varies as a function of duration of the treatment of cells with HpD and the membrane core is more susceptible to the photodynamic action of aggregated porphyrin.

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Development of Oligonucleotide Chip for Detection of Drug-Resistant Mycobacterium Tuberculosis (약제내성 결핵균의 검출을 위한 Oligonucleotide Chip의 개발)

  • Song, Eunsil;Park, Heekyung;Jang, Hyunjung;Kim, Hyomyung;Chang, Chulhun L.;Kim, Cheolmin
    • Tuberculosis and Respiratory Diseases
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    • v.55 no.1
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    • pp.41-58
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    • 2003
  • Background : The resurgence of tuberculosis and the widespread emergence of multidrug-resistant M. tuberculosis have emphasized the importance of rapid and accurate diagnostic procedures. Recently, the oligonucleotide chip has proven to be a useful tool in the rapid diagnosis of infectious diseases. The purpose of this study was to rapidly and accurately detect specific mutations in the rpoB, katG and rpsL genes associated with rifampin, isoniazid and streptomycin resistance in M. tuberculosis, respectively, using a single oligonucleotide chip. Method : For detection of drug-resistance, 7 wild-type and 13 mutant-type probes for rifampin, 2 wild-type and 3 mutant-type probes for isoniazid, and 2 wild-type and 2 mutant-type probes for streptomycin were designed and spotted onto glass slides. Fifty-five cultured samples of M. tuberculosis were amplified by PCR, and then underwent hybridization and scanning. Direct sequencing was done to verify the results from the oligonucleotide chip and to analyze the types of mutations. Result : Thirty-five cases out of 40 rifampin-resistant strains(~88%) had mutations in the rpoB gene. One case had a new mutation(D516F, GAC R TTC) and another known mutation together. Twenty cases out of 42 isoniazid-resistant strains(~50%) had mutations in the katG gene, while 7 cases out of 9 streptomycin-resistant strains(~78%) had mutations in the rpsL gene. From these results, the oligonucleotide chip was confirmed to be able to detect the most frequent mutations from the genes associated with rifampin, isoniazid and streptomycin resistance. The results proved that the drug-resistance detection probes were specific. When the results from the oligonucleotide chip and DNA sequencing were compared, the types of mutations were exactly matched. Conclusion : The diagnostic oligonucleotide chip with mutation specific probes for drug resistance is a very reliable and useful tool for the rapid and accurate diagnosis of drug resistance against rifampin, isoniazid and streptomycin in M. tuberculosis infections.

[18F]Aryl fluorides from hypervalent iodine compounds

  • Chun, Joong-Hyun;Son, Jeongmin;Park, Jun Young;Yun, Mijin
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.3 no.1
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    • pp.3-14
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    • 2017
  • Nucleophilic aromatic fluorination has been one of the most explored methods in fluorin-18 based radiochemistry. Unlike electrophilic $[^{18}F]$fluorination methods, no-carrier-added nucleophilic radiofluorination with cyclotron-produced $[^{18}F]$fluoride ion offers better specific radioactivity which is essential aspect to obtain good quality images from positron emission tomography. Contrary to amenable aliphatic radiofluorination, the development of reliable aromatic $[^{18}F]$fluorination methods has been pursued by many research groups; however, no viable method has yet been established. Recently, hypervalent iodine compound draws increasing attention as versatile radiolabeling precursor for various $[^{18}F]$fluoroarenes, since it bears the capacity to introduce fluorine-18 either on electron-deficient or electron-rich aryl ring with enhanced regiospecificity. Other classes of hypervalent iodine congeners often utilized in radiochemistry are iodylarenes, iodonium ylides, and spirocyclic iodonium ylides. Recently developed spirocyclic iodonium ylides have already been avidly employed to provide various $[^{18}F]$aryl fluorides with high labeling efficiency. This metal-free protocol would afford efficient routes, replacing the traditional approaches to $[^{18}F]$fluoroarenes, from prosthetic labeling synthons to complex PET radiotracers.