• Title/Summary/Keyword: hybridization technique

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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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Identification of Genes that are Induced after Cadmium Exposure by Suppression Subtractive Hybridization

  • 이미옥
    • Proceedings of the Korea Society of Environmental Toocicology Conference
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    • 2003.05a
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    • pp.107-107
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    • 2003
  • The heavy metal cadmium is a xenobiotic toxicant of environmental and occupational concern and it has been classified as a human carcinogen. Inhalation of cadmium has been implicated in the development of emphysema and pulmonary fibrosis, but, the detailed mechanism by which cadmium induces adverse biological effects is not yet known. Therefore, we undertook the investigation of genes that are induced after cadmium exposure to illustrate the mechanism of cadmium toxicity For this purpose, we employed the polymerase chain reaction-based suppression subtractive hybridization technique. We identified 29 different cadmium-inducible genes in human peripheral mononuclear cells, such as macrophage migration inhibitory factor, lysophosphatidic acid acyltransferase-${\alpha}$, enolase-1${\alpha}$, VEGF, Bax, neuron-derived orphan receptor-1, and Nur77, which are known to be associated with inflammation, cell survival, and apoptosis. Induction of these genes by cadmium treatment was further confirmed by semi-quantitative reverse-transcription polymerase chain reaction. Further, we found that these genes were also induced after cadmium exposure in normal human lung fibroblast cell line, WI-38, suggesting potential use of this induction profile to monitor cadmium toxicity in the lung. Next, Nur77, one of cadmium-inducible genes, was further studied since the products of Nur77 are known to be involved in the apoptotic process of lung cells. Following cadmium treatment, Nur77 gene expression was increased at protein-level in A549 cells. Consistently, the reporter containing Nur77 binding sequence was activated by 2.5-fold after exposure to cadmium in reporter gene analysis by transient transfection experiments. When the plasmid encoding dominant negative Nur77 that represses the transcriptional function of wild-type Nur77 was transfected into A549 cells, the expression of Bax was significantly reduced, suggesting that induction of Nur77 was an important process in cadmium-induced apoptosis in the cells. Cadmium induced the expression of Nur77 in vivo, confirming the relevance of the data obtained in viro. Together our results suggest that Nur77 gene expression in exposure to cadmium leads apoptosis of lung cells which may cause pathological changes in lung.

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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.

Negative HER2/neu Amplification Using Immunohistochemistry and Chromogenic in Situ Hybridization Techniques in Skin Melanoma Cases

  • Shayanfar, Nasrin;Bahari, Leila;Safaie-Naraghi, Zahra;Kamyab, Kambiz;Gheytanchi, Elmira;Rezaei, Nima
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.2
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    • pp.421-425
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    • 2015
  • Background: This study was performed to evaluate the amplification of HER-2/neu in patients with melanoma. Materials and Methods: Amplification of HER-2/neu was evaluated in a group of patients with melanoma, referred to two referral centers in Tehran, using immunohistochemistry (IHC) and chromogenic in situ hybridization (CISH) techniques. Results: Forty patients with mean age $57.9{\pm}19.5years$ were enrolled in this study. The most frequent type of melanoma was acral, while lower limbs were the most frequent sites. The amplification of HER2/neu was negative in 97.5% of patients with IHC and in 100% of patients with CISH technique. Only one case (2.5%) shows weak positive staining (+2) in IHC method. Fifty five percent of melanoma was ulcerative, and the most common stages of tumors were stages 4b and 3b. More than 47% of cases were in Clark level III, while the mean of Breslow thickness was $3.56{\pm}2.87mm$. The stage of the case that showed weakly positive staining (2+) in IHC was 4b. Conclusions: The amplification of HER2/neu biomarker was negative in patients with melanoma, using both CISH and IHC techniques.

Hydrogen Storage Capacities of MOF-5 and Microporous Carbon: Effects of Pt Loading and Hybridization (MOF-5 및 마이크로다공성 카본의 수소 저장 성능: Pt 첨가 및 하이브리드화의 영향)

  • Yunatri, Rika Tri;Suh, Dong-Jin;Suh, Young-Woong
    • Journal of Hydrogen and New Energy
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    • v.19 no.5
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    • pp.377-385
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    • 2008
  • In this study, we demonstrated that, although hydrogen molecules can be adsorbed onto the adsorbent such as MOF and MC itself, the loading of noble metal such as Pt is necessary to enhance the $H_2$ storage capacity since $H_2$ molecules can be dissociatively adsorbed on Pt metal and migrated to high-surface-area adsorbent via the primary spillover. In addition, the hybrid material have been prepared coupling MOF-5 with Pt/MC through carbon bridges formed by sucrose polymerization/carbonization. That this material showed the highest $H_2$ uptake at room temperature and about 100 bar is believed to be associated with the secondary spillover effect. Thus, such a strategy is very promising in developing $H_2$ storage technology using porous adsorbents. However, further experiments should be carried out to explore the choice of bridge carbon, the hybridization method, the dispersion technique of noble metals, etc.

A Case of Successful Pregnancy in Patient with Recurrent Spontaneous Abortion by Preimplantation Genetic Diagnosis Following IVF-ET (염색체 이상에 의한 반복 유산 환자에서 체외수정시술 및 착상전 유전진단을 통한 임신 성공 1례)

  • Jeong, Jin-Seok;Yeon, Gyu-Sun;Chae, Hee-Dong;Cheon, Yong-Pil;Kim, Chung-Hoon;Kang, Byung-Moon;Chang, Yoon-Seok;Mok, Jung-Eun
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.2
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    • pp.135-140
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    • 1998
  • It was reported that the etiologies of recurrent spontaneous abortion are immunologic factors, endocrinologic problems, anatomical abnormalities, genetic abnormalities, infection, and unexplained factors. Among those etiologic factors, genetic abnormalities occur in about 5% of the couples who experience recurrent spontaneous abortions, and most common parental chromosomal abnormality contributing to recurrent abortion is balanced translocation. The advent of in vitro fertilization (IVF), the development of skills associated with the handling of human embryo, and an explosion of knowledge in molecular biology have opened the possibility of early diagnosis of genetic disease in preimplantation embryos. Therefore preimplantation genetic diagnosis (PGD) is indicated for couples, infertile or not, at risk of transmitting a genetic disease. A case of successful pregnancy and term delivery by PGD using fluorescence in situ hybridization (FISH) technique in patient with recurrent spontaneous abortion due to balanced translocation is presented with brief review of literatures.

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Hybridization and Use Of Grapes as an Oviposition Substrate Improves the Adaptation of Olive Fly Bactrocera oleae (Rossi) (Diptera: Tephritidae) to Artificial Rearing Conditions

  • Sohel, Ahmad;Viwat, Wornoayporn;Polychronis, Rempoulakis;Emily A., Fontenot;Ul Haq, Ihsan;Carlos, Caceres;Hannes F., Paulus;Marc J.B., Vreysen
    • International Journal of Industrial Entomology and Biomaterials
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    • v.29 no.2
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    • pp.198-206
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    • 2014
  • The olive fly Bactrocera oleae (Rossi) is the key pest for olive cultivation worldwide. Substantial effort has been invested in the development of the sterile insect technique (SIT) to control this pest. One of the limitations to develop SIT technology for olive fruit fly is the low ability of wild females to lay eggs in other medium than olive fruits, and their slow adaptation to oviposition in artificial substrates. In the present study, fruit grapes were used as an alternative egg collection medium to harvest eggs and young larvae from freshly colonized wild strains originating from France, Italy, Spain and Croatia. The larvae were allowed to develop into the fruits until the second instar, before they were extracted out and further reared on a standard artificial diet. Furthermore, F1 to F4 female flies were alternatively offered wax bottles to oviposit. Finally, the performance of hybrid strains created from crosses between wild and long colonised flies was assessed. The results showed that females of all 4 wild strains readily oviposited eggs in grapes and from the F2 generation onward, females from all strains were adapted to laying eggs in wax bottles. No difference was observed in eggs and pupae production among all strains tested. The findings are discussed for their implications on SIT application against olive fruit fly.

Potato breeding via protoplast fusion (원형질체 융합을 이용한 감자 육종)

  • Cho, Kwang-Soo;Park, Tae-Ho
    • Journal of Plant Biotechnology
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    • v.41 no.2
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    • pp.65-72
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    • 2014
  • Plant cells from which the cell walls have been enzymatically or mechanically removed are called protoplasts. The protoplasts are theoretically totipotent and can be used as sources of somatic cell fusion in practical breeding programs. Wild Solanum species have often been used as sources of important agricultural traits including diverse disease resistance. However, they cannot often be directly applied to breeding programs due to their sexual incompatibility with S. tuberosum. Somatic hybridization via protoplast fusion is one of the ideal methods to overcome this limitation and to introgress certain traits into S. tuberosum. This technique has still widely been used in potato since the first fusion was reported in 1970s. Therefore, this review highlights general perspectives of protoplast fusion and discusses the application of protoplast fusion in potato breeding.

Optimal Environmental and Economic Operation using Evolutionary Computation and Neural Networks (진화연산과 신경망이론을 이용한 전력계통의 최적환경 및 경제운용)

  • Rhee, Sang-Bong;Kim, Kyu-Ho;You, Seok-Ku
    • The Transactions of the Korean Institute of Electrical Engineers A
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    • v.48 no.12
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    • pp.1498-1506
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    • 1999
  • In this paper, a hybridization of Evolutionary Strategy (ES) and a Two-Phase Neural Network(TPNN) is applied to the optimal environmental and economic operation. As the evolutionary computation, ES is to search for the global optimum based on natural selection and genetics but it shows a defect of reducing the convergence rate in the latter part of search, and often does not search the exact solution. Also, neural network theory as a local search technique can be used to search a more exact solution. But it also has the defect that a solution frequently sticks to the local region. So, new algorithm is presented as hybrid methods by combining merits of two methods. The hybrid algorithm has been tested on Emission Constrained Economic Dispatch (ECED) problem and Weighted Emission Economic Dispatch (WEED) problem for optimal environmental and economic operation. The result indicated that the hybrid approach can outperform the other computational efficiency and accuracy.

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Diagnosis of human genetic mutations based on DNA microarray technology

  • Park, Hyun-Gyu
    • 한국생물공학회:학술대회논문집
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    • 2005.04a
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    • pp.17-17
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    • 2005
  • In this presentation, we will discuss several recent achievements developed in my laboratory for microarray-based diagnosis of human genetic mutations including HNF-1 and BRCA1 mutations. To determine the presence of the genetic mutations in a human sample, we prepared allele-specific oligonucleotide chips from selected mutation sites and generated target probes using a tow-step method for Cy-3 DNA $samples^{1)}$ or in vitro transcription of promoter-tagged PCR products for Cy-3 RNA $samples^{2)}$. Hybridization of the target probes to the chips successfully identified all of the genotypes for the tested sites. For more reliable diagnosis, we also employed single base extension (SBE) reaction and zip-code microarray technique for our strategy. Particularly we developed an efficient PNA zip-code microarray for the detection of $HNF-1{\alpha}$ $mutations^{3)}$. Using multiplex SBE reactions and zip-code strategy, we were able to correctly diagnose several mutation sites in exon 2 of $HNF-1{\alpha}$ with a wild-type and mutant including a MODY3 patient. These works represent successful applications of DNA microarray technology for the diagnosis of human genetic mutations.

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