• Title/Summary/Keyword: isolation techniques

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Novel Vectors for the Convenient Cloning and Expression of In Vivo Biotinylated Proteins in Escherichia coli

  • Cho, Eun-Wie;Park, Jung-Hyun;Na, Shin-Young;Kim, Kil-Lyong
    • BMB Reports
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    • v.32 no.5
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    • pp.497-501
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    • 1999
  • Biotinylation of recombinant proteins is a powerful tool for the detection and analysis of proteins of interest in a large variety of assay systems. The recent development of in vivo biotinylation techniques in E. coli has opened new possibilities for the production of site-specifically biotinylated proteins without the need for further manipulation after the isolation of the recombinantly expressed proteins. In the present study, a novel vector set was generated which allows the convenient cloning and expression of proteins of interest fused with an N-terminal in vivo biotinylated thioredoxin (TRX) protein. These vectors were derived from the previously reported pBIOTRX vector into which was incorporated part of the pBluescript II+phagemid multiple cloning site (MCS), amplified by PCR using a pair of sophisticated oligonucleotide primers. The functionality of these novel vectors was examined in this system by recombinant expression of rat transforming growth factor-$\beta$. Western-blot analysis using TRX-specific antibodies or peroxidase-conjugated streptavidin confirmed the successful induction of the fusion protein and the in vivo conjugation of biotin molecules, respectively. The convenience of molecular subcloning provided by the MCS and the effective in vivo biotinylation of proteins of interest makes this novel vector set an interesting alternative for the production of biotinylated proteins.

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A Study on the Contents of Fundamental Nursing Practice (기본간호학 실습교육현황과 실습내용 개선을 위한 조사 연구)

  • 이영휘;서화숙
    • Journal of Korean Academy of Nursing
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    • v.28 no.3
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    • pp.650-661
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    • 1998
  • The purpose of this study was to provide fundamental data for determining contents of Fundamental Nursing Practice and developing desirable bedside nursing techniques for clinical nursing areas. Subjects for this study were 86 nurses who were employees of two university hospitals located in Seoul and a district area. Data were collected by questionnaires. Items of Fundamental Nursing Practice were classified into 72 items according to the result from content analysis of 9 textbooks of Fundamental Nursing. The results are as follows : 1) Items which above 80% of respondents practiced during the school inside practice or clinical nursing practice course were axillary temperature measurement, radial pulse measurement, respiratory rate measurement, application of oral hygiene, hand-washing technique, application of hot and cold bags, intramuscular injection technique, open bed-making, soap enema method, application of nelaton catheterization and oral and nasal suction methods. 2) Above 90% of respondents replied that all items except temperature measurement and bed-making were requisite contents for Fundamental Nursing Practice. Above 10% of respondents replied oral and rectal temperature measurement and bed-making were unnecessary content. 3) Above 90% of respondents replied that operating methods of all items except isolation technique, admission and discharge procedures, and retention enema in the Fundamental Nursing Practice course and clinical situation were consistent. The main reasons that respondents did not apply methods which they learned in the Fundamental Nursing Practice course to the clinical situation were 'insufficient time', 'colleagues were using different methods', 'insufficient supply of instuments' or 'inappropriate appliances'.

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Two New Phenolic Compounds from the Fruiting Bodies of Ganoderma tropicum

  • Hu, Li-Li;Ma, Qing-Yun;Huang, Sheng-Zhuo;Guo, Zhi-Kai;Guo, Jian-Chun;Dai, Hao-Fu;Zhao, You-Xing
    • Bulletin of the Korean Chemical Society
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    • v.34 no.3
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    • pp.884-886
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    • 2013
  • Chemical investigation of the fruiting bodies of Ganoderma tropicum led to the isolation of two new phenolic compounds, ganodermatropins A (1) and B (2). Their structures were elucidated by spectroscopic techniques (MS, 1D and 2D NMR). Ganodermatropin A exhibited antimicrobial activity against Staphylococcus aureus.

Purification and Structural Characterization of Glycolipid Biosurfactants from Pseudomonas aeruginoas YPJ-80

  • Park, Oh-Jin;Lee, Young-Eun;Cho, Joong-Hoon;Shin, Hyun-Jae;Yoon, Byung-Dae;Yang, Ji-Won
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.3 no.2
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    • pp.61-66
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    • 1998
  • Glycolipids produced by Pseudomonas aeruginosa YPJ-80 were characterized by chromatographic and spectorscopic techniques as a mixture of two rhamnolipids. For recovery of glycolipids from the culture broth, various isolation methods including ultrafiltration, adsorption and solvent extraction were compared. Ultrafiltration showed the best results in terms of glycolipids recovery. Further purification for spectroscopic analysis was carried out by adsorption chromatography and preparative thin layer chromatography. From the spectroscopic analysis, such as IR spectroscopy. FAB-MS, 1H-NMR and 13C-NMR and hydrolysis analysis, the glycolipids were identified as L-${\alpha}$-rhamnopyranosyl-${\beta}$-hydroxydecanoly-${\beta}$-hydroxydecanoate and 2-O-${\alpha}$-L-rhamnopyranosyl-${\alpha}$-L-rhamnopyranosyl-${\beta}$-hydroxydecanoyl-${\beta}$-hydroxydecanoate. Monorhamnolipid and dirhamnolipid lowered the surface tension of water to 28.1 mN/M and 29.3 mN/m, respectively.

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Design Structure Matrix: A Model Proposal and Implementation on Harbor and Building Design Project

  • Akram, Salman;Kim, Jeonghwan;Pi, Seungwoo;Seo, Jongwon
    • Korean Journal of Construction Engineering and Management
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    • v.14 no.1
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    • pp.144-152
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    • 2013
  • Design is an iterative, generative, and multidisciplinary process by its nature. Iteration occurs often in most of the engineering design and development projects including construction. Design iterations cause rework, and extra efforts are required to get the optimal sequence and to manage the projects. Contrary to simple design, isolation of the generative iterations in complex design systems is very difficult, but reduction in overall iterations is possible. Design depends upon the information flow within domain and also among various design disciplines and organizations. Therefore, it is suggested that managers should be aware about the crucial iterations causing rework and optimal sequence as well. In this way, managers can handle design parameters related to such iterations pro-actively. There are a number of techniques to reduce iterations for various kinds of engineering designs. In this paper, parameter based Design Structure Matrix (DSM) is chosen. To create this DSM, a survey was performed and then partitioned using a model. This paper provides an easy approach to those companies involved in or intend to be involved in "design and build projects".

Polarization Splitter Made of the Side-Polished Fiber Coupler Including a thin Metal Intermediate Layer (얇은 중간 금속층을 포함한 측면 연마 광섬유 결합기를 이용한 편광 분리기)

  • 김광택;이준옥
    • Journal of the Institute of Electronics Engineers of Korea SD
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    • v.40 no.9
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    • pp.651-657
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    • 2003
  • We have demonstrated an in-line polarization splitter based on a side-polished fiber coupler including a thin metal intermediate layer. The experimental results revealed that the metal layer with proper thickness prevents TE polarization component from optical coupling between two contacted side-polished fibers, whereas it allows TM polarization component to the coupling. The design and fabrication techniques about the polarization splitter exploiting the side-polished fibers have been presented. The fabricated polarization splitter exhibited 18dB and 23dB of isolation ratio for TE polarization and TM polarization, respectively. The measured insertion loss for TE and TM polarization was 0.7dB and 1.3dB, respectively.

Isolation and Differential Expression of an Acidic PR-1 cDNA Gene from Soybean Hypocotyls Infected with Phtophthora sojae f. sp. glycines

  • Kim, Choong-Seo;Yi, Seung-Youn;Lee, Yeon-Kyung;Hwang, Byung-Kook
    • The Plant Pathology Journal
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    • v.16 no.1
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    • pp.9-18
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    • 2000
  • Using differential display techniques, a new acidic pathogenesis-related (PR) protein-1 cDNA (GMPRla) gene was isolated from a cDNA library of soybean (Glycinemax L.Merr, cultivar Jangyup) hypocotyls infected by Phytophthora sojae f. sp. glycines. The 741 bp of fulllength GMPRla clone contains an open reading frame of 525 nucleotides encoding 174 amino acid residues (pI 4.23) with a putative signal peptide of 27 amino acids in the N-terminus. Predicted molecular weight of the protein is 18,767 Da. The deduced amino acid sequence of GMPRla has a high level of identity with PR-1 proteins from Brassica napus, Nicotiana tabacum, and Sambucus nigra. The GMPRla mRNA was more strongly expressed in the incompatible than the compatible interaction. The transcript accumulation was induced in the soybbean hypocotyls by treatment with ethephon or DL-$\beta$-amino-n-butyric acid, but not by wounding. In situ hybridization data showed that GMPRIa mRNAs were usually localized in the vascular bundle of hypocotyl tissues, especially phloem tissue. Differences between compatible and incompatible interactions in the timing of GMPRla mRNA accumulation were remarkable, but the spatial distribution of GMPRla mRNA was similar in both interactions. However, more GMPRla mRNA was accumulated in soybean hypocotyls at 6 and 24 h after inoculation.

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Phenolic Components from the Fruits of Pourthiaea villosa (윤노리나무 과실의 페놀성 성분)

  • Lee, Hyun Jin;Ahn, Dalrae;Lee, Eun Byeol;Lee, Tae Gwan;Kim, Dae Keun
    • Korean Journal of Pharmacognosy
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    • v.44 no.1
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    • pp.16-21
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    • 2013
  • The fruits of Pourthiaea villosa were extracted with methanol and its extract was fractionated with n-hexane, methylene chloride, ethyl acetate and n-butanol. Repeated column chromatography of silica gel, sephadex LH-20 and HPLC led to the isolation of nine phenolic compounds from ethyl acetate soluble fraction. The chemical structures were elucidated as kaemferol-3-O-${\beta}$-D-glucopyranoside (astragalin) (1), isorhamnetin-3-O-${\beta}$-D-glucopyranoside (2), kaempferol-3-O-${\beta}$-D-xylopyranosyl($1{\rightarrow}2$)-${\alpha}$-L-rhamnopyranoside (3), caffeic acid (4), quercetin-3-O-${\beta}$-D-xylopyranosyl($1{\rightarrow}2$)-a-L-rhamnopyranoside (5), quercetin-3-O-${\beta}$-D-xylopyranosyl($1{\rightarrow}2$)-${\beta}$-D-glucopyranoside (6), quercetin-3-O-${\beta}$-D-xylopyranosyl($1{\rightarrow}2$)-${\beta}$-D-galactopyranoside (7), quercetin-3-O-${\alpha}$-L-rhamnopyranoside (quercitrin) (8), and kaempferol-3-O-${\alpha}$-L-rhamnopyranoside (afzelin) (9) by spectroscopic techniques. These compounds were isolated from this plant for the first time.

Displacement aging component-based stability analysis for the concrete dam

  • Huang, Xiaofei;Zheng, Dongjian;Yang, Meng;Gu, Hao;Su, Huaizhi;Cui, Xinbo;Cao, Wenhan
    • Geomechanics and Engineering
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    • v.14 no.3
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    • pp.241-246
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    • 2018
  • The displacement monitoring data series reconstruction method was developed under equal water level effects based on displacement monitoring data of concrete dams. A dam displacement variation equation was set up under the action of temperature and aging factors by optimized analysis techniques and then the dam displacement hydraulic pressure components can be separated. Through the dynamic adjustment of temperature and aging effect factors, the aging component isolation method of dam displacement was developed. Utilizing the isolated dam displacement aging components, the dam stability model was established. Then, the dam stability criterion was put forward based on convergence and divergence of dam displacement aging components and catastrophe theory. The validity of the proposed method was finally verified combined with the case study.

Identification of Non-mutans Streptococci Organisms in Dental Plaques Recovering on Mitis-Salivarius Bacitracin Agar Medium

  • Yoo So Young;Kim Pyung Sik;Hwan Ho Keel;Lim Seong Hoon;Kim Kwang Won;Choe Son Jin;Min Byung Moo;Kook Joong Ki
    • Journal of Microbiology
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    • v.43 no.2
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    • pp.204-208
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    • 2005
  • The objective of this study was to both isolate and identify non-mutans streptococci organisms (non­MSO) from dental plaques recovered on mitis-salivarius sucrose bacitracin agar (MSB) plates. The dental plaque samples, which had been collected from 63 human subjects, were diluted and plated on MSB. The bacteria growing on the MSB plates were then identified with biochemical tests, as well as with 16S rDNA cloning and sequencing techniques. Our data indicated that bacteria from 30 subjects had been recovered on the MSB plates. Among the 21 typical colonies selected from the 30 subjects, 12 colonies, derived from 10 subjects, were identified as non-MSO. These 12 colonies were determined to be Streptococcus anginosus (8 colonies), S. sanguinis (1 colony), and Pantoea agglomerans (3 colonies). These results strongly suggest that a new selective medium will be required for the reliable isolation of mutans streptococci.