• Title/Summary/Keyword: Strand method

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Complementary DNA Cloning of Genomic RNA in Orchid Strain of Tobacco Mosaic Virus

  • Won Mok Park
    • Journal of Plant Biology
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    • v.37 no.3
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    • pp.349-355
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    • 1994
  • Viral RNA was extracted from a purified orchid strain of tobacco mosaic virus (TMV-O) from Cymbidium "Grace Kelly". Polyadenylated viral RNAs were primed with Not I-oligo (dT) primer-adapter. First-strand cDNAs were reversely transcribed by Moloney murine leukaemia virus reverse transcriptase (RNAse H-), and then second-strand cDNAs were synthesized by RNase H and DNA polymerase I. The resulting double-stranded cDNAs were ligated into pSPORT1 vector and transformed into competent E. coli strain JM109 cells. The size of cDNAs within the recombinant plasmids was ranging from 0.9 to 3.9 kb. Among the selected clones, pTMO-0205 and -0210 covered the 3' half and the 5' half of the viral genomic RNA, respectively, which were covering more than 99% of the viral genemo size based on sequencing analysis. Two cDNA fragments which were 3.1 kb BamHI and NotI fragement released from pTMO-0.205 and 3.3 kb SalI and BamHI fragment released from pTMO-0210 were ligated with T4 DNA ligase. The clone was almost entire length, lacking only 31 nucleotides from the 5' terminus based on the sequencing result. This method was shown to be efficiently applicable to other plant viral gnomic RNA for the construction of cDNA.n of cDNA.

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Molecular Cloning of H-Y Antigen Gene III. Construction of Mouse Testis cDNA Library and Screening of H-Y Ag Gene (H-Y 항원 유전자의 클로닝에 관한 연구 III. 생쥐정소 cDNA Library 구성과 유전자의 검색)

  • 이정렬;김창규;김종배
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.43-48
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    • 1993
  • These experiments were carried out to construct mouse testis cDNA library and to to seen H-Y Ag gene. Mouse testis was obtained from BALB/c inbreed mouse that was after-born 1 week. Isolation of mouse testis total RNA was carried out by guanidum/cesium choloride, poly(A+) mRNAs were purified by oligo d(T)-cellulose chromatography method. To investigate protein synthesis activity, in-vitro translation carried out by total RNA and poly(A+) mRNA. The products of in-vitro translation were identified in 12.5% PAGE. Single strand DNA and double strand DNA were synthesized from poly(A+) mRNA and purified using phenol/chloroform/isoamylalcohol. Synthesized cDNA was combined with cohesive Eco RI polylinker, its recombination efficiencies were identified by X-gal and IPTG. In the cDNA library, 1$\times$107 phagemids were screened with 32P labelled probe. Hybridization were carried on $65^{\circ}C$ for 16~20hours. And 1$\times$106 phagemids were screened with rabbit-anti-H-Y. In former, select 5 positive clones, and later, 1 positive clone. Its southern blot analysis showed various size of insert cDNA from 0.7kb to 3kb.

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Numerical study of mono-strand anchorage mechanism under service load

  • Marceau, D.;Fafard, M.;Bastien, J.
    • Structural Engineering and Mechanics
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    • v.18 no.4
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    • pp.475-491
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    • 2004
  • Anchorage devices play an important role in post-tensioned bridge structures since they must sustain heavy loads in order to permit the transfer of the prestressing force to the structure. In external prestressing, the situation is even more critical since the anchorage mechanisms, with the deviators, are the only links between the structure and the tendons throughout the service life of the structure. The behaviour of anchorage devise may be studied by using the finite element method. To do so, each component of the anchorage must be adequately represented in order to approximate the anchor mechanism as accurately as possible. In particular, the modelling of the jaw/tendon device may be carried out using the real geometry of these two components with an appropriate constitutive contact law or by replacing these components by a single equivalent. This paper presents the numerical study of a mono-strand anchorage device. The results of a comparison between two different representations of the jaw/tendon device, either as two distinct components or as a single equivalent, will be examined. In the double-component setup, the influence of the wedge configuration composing the jaw, and the influence of lubrication of the anchor, will be assessed.

A Reconstruction of Probability Unit of Elementary Mathematics Textbook Based on Freudenthal's Reinvention Method (Freudenthal의 재발명 방법에 기초한 제7차 초등수학교과서 확률 단원 재구성)

  • Kang, Ho-Jin;Kang, Heung-Kyu
    • Journal of Elementary Mathematics Education in Korea
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    • v.12 no.1
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    • pp.79-100
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    • 2008
  • Freudenthal has advocated the reinvention method. In that method, the pupils start with a meaningful context, not ready-made concepts, and invent informative method through which he could arrive at the formative concepts progressively. In many face the reinvention method is contrary to the traditional method. In traditional method, which was named as 'concretization method' by Freudenthal, the pupils start with ready-made concepts, and applicate this concepts to various instances through which he could arrive at the understanding progressively. Through analysis, it turns out that Korea's seventh elementary mathematics textbook is based on concretization method. In this thesis, first of all, I will reconstruct probability unit of seventh elementary textbook according to Freudenthal's reinvention method. Next, I will perform teaching experiment which is ruled by new lesson design. Lastly, I analysed the effects of teaching experiment. Through this study, I obtained the following results and suggestions. First, the reinvention method is effective on the teaching of probability concept and algorithm. Second, in comparison with current textbook strand, my strand which made probability concept go ahead and combinatorics concept let behind is not deficiency. Third, tree diagram is effective matrix which contribute to formalization of combinatorics calculation. Lastly, except for fraction, diverse representation of probability, for example percentage or informal ratio expression must be introduced in teaching process.

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Simultaneous Detection of 10 Foodborne Pathogens using Capillary Electrophoresis-Based Single Strand Conformation Polymorphism

  • Oh, Mi-Hwa;Hwang, Hee-Sung;Chung, Bo-Ram;Paik, Hyun-Dong;Han, Sang-Ha;Kang, Sun-Moon;Ham, Jun-Sang;Kim, Hyoun-Wook;Seol, Kuk-Hwan;Jang, Ae-Ra;Jung, Gyoo-Yeol
    • Food Science of Animal Resources
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    • v.32 no.2
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    • pp.241-246
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    • 2012
  • This report outlines the development of a rapid, simple, and sensitive detection system for pathogenic bacteria using a capillary electrophoresis-based, single strand conformation polymorphism (CE-SSCP) combined with PCR. We demonstrate that this method, used with primers targeting the V4 region of the16S rRNA gene, is capable of the simultaneous detection of 10 microbes that could be associated with foodborne illness, caused by animal-derived foods: Salmonella enterica, Listeria monocytogenes, Escherichia coli O157:H7, Campylobacter jejuni, Staphylococcus aureus, Bacillus cereus, Clostridium perfringens, Yersinia enterocolitica, Vibrio parahaemolyticus, and Enterobacter sakazakii. The traditional detection techniques are time-consuming and labor-intensive, due to the necessary task of separate cultivation of each target species. As such, the CE-SSCP-PCR method, that we have developed, has the potential to diagnose pathogens rapidly, unlike the traditional technique, in order to prevent foodborne illness in a much more efficient manner.

Analysis of p53 Somatic Mutation in Head and Neck Cancer Using Denaturing High Performance Liquid Chromatography(DHPLC) (두경부 종양에서 DHPLC를 이용한 p53체세포 돌연변이 검출 연구)

  • Kim, Kwang-Youl;Park, Sang-Bum;Han, Sang-Man;Nam, Youn-Hyoung;Jang, Won-Cheoul
    • Journal of the Korean Chemical Society
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    • v.48 no.1
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    • pp.33-38
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    • 2004
  • Mutation of p53 tumor suppressor gene in HNSCC (head and neck squamous cell carcinoma) has been proposed high rate. We extracted genomic DNA from 50 head and neck cancer. The DNA was amplified by PCR at exon 5-8 in p53 tumor suppressor gene. We have compared single strand conformation polymorphism (SSCP) and denaturing high performance liquid chromatography (DHPLC) method for analysis of p53 somatic mutation. As a result, 16 deleted mutations (32%) were detected by SSCP analysis and 17 deleted mutations (34%) were detected by DHPLC analysis at exon 8. All of 17 mutations were proved by sequencing. We conclude that DHPLC is a fast and simple screening method rather than SSCP analysis.

Analysis of the Manufacturing Techniques for the KwangDahoe Tying on the Sword in Joseon Dynasty (조선시대 도검 패용 광다회의 제작기법 분석)

  • Baek, Je-Sun;Chung, Kwang-Young
    • MUNHWAJAE Korean Journal of Cultural Heritage Studies
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    • v.50 no.3
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    • pp.64-87
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    • 2017
  • Dahoe is a traditional braid in Joseon Dynasty. There are many Dahoe artifacts, which can give us a glimpse of the costume, culture, and life of that era. The study of Dahoe is necessary in various fields, but it is difficult because this manufacturing technique has been passed down from hand to hand. Few studies have been done so far. This research examines the manufacturing technique of KwangDahoe which is passed down by Maedeupjang (Decorative Knotting) and is generally used for knotting and/or tying objects. The main characteristic of TieKwangDahoe, made through the same method as WonDahoe, is the square hole in the middle. It was impossibile to remake the original braid because there is no confirmed number of the strand. Especially it is very difficult to do conservation and restoration on serious degradation state of the fiber. Therefore, it is necessary to analyze the non-destructive manufacturing techniques method for Dahoe and assess their applicability. First, we analyzed the artifacts' manufacturing technique based on the database of the Dahoe's manufacturing technique. In order to do that, we undertook schematization, restoration, morphological analyzation of the Dahoe. And then, X-ray CT scans were performed to improve the reliability of the DB. These results of scanning were interpreted based on the manufacturing technique. The selected Tie-KwangDahoe on the sword for the study are artifacts including artistic value and symbolism in Joseon Dynasty. Based on the analysis of the manufacturing technique, we found that both artifacts were made of 20-strand braid of single cross according to the length-scale measure. It was manufactured using 8-strand on left-right side, 12-strand on front-back side by the braiding manufacturing technique method Finally, this research suggests non-destructive analysis method of Dahoe's manufacturing technique is based on the database and the analysis results. I hope this research can be useful in various professional fields of Dahoe in the future. Moreover, I hope this can be of any help in preserving Korean cultural heritage.

An Estimation on Application of Slope Stabilization Method Using PC Strand Cable (케이블을 이용한 사면안정 공법 적용성 평가)

  • Kim, Hong-Taek;Baek, Seung-Cheol;Yoo, Chan-Ho;Jang, Chung-Gil;Min, Kyung-Nam
    • Proceedings of the Korean Geotechical Society Conference
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    • pp.984-991
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    • 2008
  • In this study, to evaluate the slope and tunnel stabilization method using pc cable with bulbed. To estimate the application of tunnel support using field tests and numerical analysis results. The reinforcement effects of slope stabilization method reinforced by PC cables were estimated compared with conventional soil nail system that reinforce the slope using rebar.

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New Methods for Assessing Liquefaction Potential Based on the Characteristics of Material (재료의 역학적 거동특성에 기초한 액상화 평가방법)

  • Kim, Gyeong-Hwan;Park, In-Jun;Kim, Su-Il
    • Geotechnical Engineering
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    • v.14 no.5
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    • pp.205-218
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    • 1998
  • The purpose of this study is to develop and utilize new assessment of liquefaction potential based on DSC(disturbed state concept) and dissipated energy concept. The term liquefaction has suddenly loses its shear strength and behaves like a fluid. Liquefaction has been a source of a major damage during severe earthquake. In this study, the cyclic undrained behavior of Joomoonjin strand is investigated by using an automates triaxial testing device(C. K. Chan type). In order to assess liquefaction potential of saturated strand, DSC method and energy method are applied for the experimental data. The use of DSC method and energy method to define the liquefaction potential is verified through laboratory testis of cyclic triaxial test on saturated sand specimens. Based on the analytical results of DSC method, the relationship between the factor affecting liquefaction characteristics(Dr) and physical properties of the saturated santa(fs and D.) is found. Based on the analytical results of energy method, it is found that the initial liquefaction of rand is related to the significant change in the dissipated energy. Finally, it is shown that the DSC method and energy method can capture the liquefaction mechanism.

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Anterior Cruciate Ligament Double Bundle Reconstruction with Hamstring Tendon Autografts - Technical Notes (자가 슬괵건을 이용한 전방 십자 인대 이준 다발 재건술 - 수술 술기 -)

  • Ahn, Jin-Hwan;Lee, Sang-Hak;Ahn, Hyung-Kwon;Kang, Hong-Jae
    • Journal of the Korean Arthroscopy Society
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    • v.9 no.2
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    • pp.222-231
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    • 2005
  • Purpose: This article describes a double-bundle ACL reconstruction technique using a five-strand hamstring tendon autograft with conventional anteromedial bundle reconstruction and additional posterolateral bundle reconstruction. Operative technique: For the tibial tunnel, the conventional single tunnel technique is performed and for the femoral tunnel, the double tunnel technique is performed with the anteromedial and posterolateral bundle. After minimal notchplasty, the anteromedial femoral tunnel is prepared with leaving one milimeter of posterior femoral cortex within the over-the-top, which if positioned at the 11-o'clock orientation for the right knee or at the 1-o'clock position for the left knee. The posterolateral femoral tunnel that is located 5 to 7 mm superior to the inner margin of the lateral meniscus anterior horn at $90^{\circ}$ of flexion is prepared with tile outside-in technique using a 4.5 cannulated reamer. The graft material for the double bundle reconstruction is made of the conventional four-strand hamstring autograft in the anteromedial bundle and of a single-strand semitendinosus tendon in the posterolateral bundle. The anteromedial bundle is fixed with using a rigid fix system on the femoral side and the posterolateral bundle is fixed to tie with the miniplate from the outside femur. Then, with the knee in $10^{\circ}\;to\;20^{\circ}$ of flexion, a bioabsorbable screw is simultaneously applied to achieve tibial fixation with tensioning of both bundles. Conclusion: A double bundle reconstruction with five-strand hamstring autograft, which is designed with a favorable conventional anteromedial bundle and an additional posterolateral bundle to restore rotation stability, seems to be a very effective method for the treatment for ACL instabilities.

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