• Title/Summary/Keyword: marker exchange

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Marker-Exchange Mutagenesis of Pectate Lyase Gene in Rhizobium fredii (Rhizobium fredii Pectate Lyase 유전자의 Marker-Exchange 변이)

  • 정민화;박용우;윤한대
    • Microbiology and Biotechnology Letters
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    • v.19 no.3
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    • pp.222-227
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    • 1991
  • Rhizobium fredii USDA193 is one of the causal organism for root nodule formation in soybean (peking). Previously we cloned the pectate lyase gene (SY1) of R. fredii USDA193. The $pel^-$ mutants (SY1$\Omega$ and SY1$\Omega$1) of SY1 were obtained using the in vitro insertional omega mutagenesis of RpelB (of Rhizobium pel) and fill-in reaction of RpelE (of Rhizobium pel) gene respectively, and we constructed two mutants (R, fredii USDA193$\Omega$ and R. fredii USDA193$\Omega$1) in pectate lyase function by marker-exchange with pe1B::$\Omega$ and R. fredii USDA193 strain (rif). The pectate lyase activity of two pel- mutant of R. fredii USDA193 was determined by spectrophotometric method. However, all pectate lyase activity of these mutants was not lost upon the mutagenesis by marker-exchange. This suggests that other pectate lyase genes may be present on the plasmid or the chromosome of R. fredii. As yet we do not have evidence linking RpelB and RpelE genes of R. fredii directly to the early nodulation process.

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Determination of Stress Profiles by Fractography in Single Ion-exchanged Glass (파괴분석을 이용한 단일이온교환된 유리의 응력 형성 관찰)

  • 이회관;강원호
    • Journal of the Microelectronics and Packaging Society
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    • v.10 no.4
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    • pp.61-64
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    • 2003
  • The fractography was used to observe the stress profile on a single ion-exchanged glass. In the processing, the temperature was varied during the ion exchange, and then the stress profile on the single ion-exchanged glass was shifted from glass surface to slightly below the glass surface. The hackle mark and mirror surface were varied according to process conditions, and the glass strength was increased with increasing the number of hackle markers. In case of breaking the stress profile by using indenter, the breaking property was similar to the annealed glass.

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An Approach for Reducing Carbon-14 Stack Emissions via Optimal Use of Ion Exchang Resins at CANDU Plant

  • Sohn, Wook;Chi, Jun-Ha;Kang, Duk-Won
    • Proceedings of the Korea Society for Energy Engineering kosee Conference
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    • 2003.05a
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    • pp.445-455
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    • 2003
  • Relatively high carbon-14 emissions, which occurred at PHWR Plant during 1998 and 1999, made the site staff to implement several operational improvements: 1) the frequency and volume of the moderator cover gas purging were reduced through increased $O_2$ additions to the cover gas, 2) the 'old' resin columns were not used during re-start of the reactor from outage, 3) efforts were made to minimize air ingress, 4) the maximum service time of moderator ion-exchange columns were restricted to about 80 days. Through the improvements, the carbon-14 emission from each PHWR reactor returned to the normal levels during the remainder of 1999 and during 2000. We carried out a special surveillance at W-1 and W-3 from September 2001 to August 2002 to properly evaluate ways to optimize the use of moderator ion exchange resins from a C-14 perspective. The surveillance showed that only data that provided an operational marker for deciding when to remove the IX-resin column is an observed increase in the C-14 stack emissions themselves. Also, it is shown that any increase over the rate of 0.4 Ci $month^{-1}$ for two consecutive weeks may be the indication for an ion-exchange resin column change, especially if the IX-resin column has been in service for more than 80 days.

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Discourse Markers in EFL Learners' Turn-Taking through Computer-Mediated Communication (CMC)

  • Hahn, Hye-Ryeong
    • English Language & Literature Teaching
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    • v.13 no.4
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    • pp.33-58
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    • 2007
  • The present paper aims at comparing the two modes of CMC - synchronous and asynchronous - in terms of discourse markers used in turn-initial positions. It further attempts to examine the viability and limitations of these two modes of CMC in fostering EFL learners' face-to-face conversation skills. For these purposes, the present study analyzed 33 Korean EFL learners' Web chat and E-mail exchange data. Discourse markers in the participants' Web chat transcripts and those in their E-mail transcripts were identified and then compared in terms of their frequency and functions. The analysis revealed that the participants show difference in their preference for discourse markers depending on the modes of CMC. Also, the functions of discourse markers used for Web chat showed were strikingly different from those for e-mail. Especially, e-mail discourse markers revealed greater discrepancy from the markers in face-to-face conversation. The differences were found to be attributable to the time factor involved with the turn-taking systems of the two modes of CMC, especially the degree of instantaneousness in their turn-taking. Findings suggest that the turn taking skills and discourse marker use in CMC is not applicable to face-to-face conversation contexts. Pedagogical implications are discussed.

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Clonig of CM-cellulase Gene of Rhizobium meliloti TAL1372 in Escherichia coli (Rhizobium meliloti TAL1372에서 섬유소분해효소 유전자 클로닝)

  • Park, Yong-Woo;Lim, Sun-Teak;Kang, Kyu-Young;Yun, Han-Dae
    • Applied Biological Chemistry
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    • v.38 no.4
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    • pp.313-319
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    • 1995
  • The involvement of the cell-wall degrading enzymes in Rhizobium has long been an unsolved question about the infection process in the formation of root nodule. To assess the contribution of the cellulase to the nodulation of rhzobia, here we report the production of cellulase from R. meliloti TAL1372 which degrade carboxymethylcellulose (CMC) model substrate with CMC-plate method. We constructed a genomic library by cloning Sau3A-digested genomic DNA from R. meliloti TAL1372 into the BamHI site of the cosmid vector pLAFR3. Out of more than one thousand transductants of E. coli, one clone (pRC8-71) had CM-cellulase activity and contained pLAFR3 cosmid with 30 kb insert of R. meliloti DNA The product of CM-cellulase gene was analyzed by native PAGE. About 45 kD protein was considered to be a product of the gene. Tn5 mutagenesis reveals that the structural gene located in a ca. 3 kb KpnI fragment. The cellulase-minus mutants of R. meliloti TAL1372 were obtained by Tn5 mutagenesis of pRC8-71 and marker exchange techniques. Analyses of the nodulation ability of these Tn5 mutants showed that the CM-cellulase gene of R. meliloti TAL1372 may be involved in early nodulation development on alfalfa (Medicago satiua).

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NDP Kinases Suppressed Bax-Dependent Apoptosis in Yeast System

  • K. C. Hwang;D. W. Ok;D. N. Kwon;H. K. Shin;Kim, J. H.
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.52-52
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    • 2001
  • Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic Many nucleoside diphosphate (NDP) kinases are ubiquitous enzymes responsible for the exchange of ${\gamma}$-phosphates between tri- and diphosphonucleosides. The catalytic reaction follows a ping-pong mechanism in which the enzyme is transiently phosphorylated on a histidine residue conserved in all nucleoside diphosphate kinases. Beside their role in nucleotide synthesis, these enzymes present additional functions, possibly independent of catalysis, in processes such as differentiation, cell growth, tumor progression, metastasis and development. To clone murine nm23-M5, several expressed sequence tags (ESTs) of the GenBank data base, selected according to their homology to nm23-H5 cDNA, reconstituted a complete open reading frame (GenBank AF222750). To test whether murine NDPKs (1, 2, 3, 4, 5, and 6) can inhibit Bax-mediated toxicity in yeast, co-transformation was performed respectively. The yeast S.cerevisiae was transformed with a copy expression plasmid containing the histidine selection marker and expressing murine Bax under the control of a galactose-inducible promoter. Several clones were selected and found to be growth inhibited when Bax expression was induced with galactose. A representative clone was transformed again with a copy expression plasmid containing the tryptophane selection marker and expressing either murine Bcl-xL or NDPK under the control of a galactose-inducible promoter. Several subclones of the double-transformants were selected and characterized. The ability of Bcl-xL and NDPKs to suppress Bax-mediated toxicity was determined by growing yeast cells overnight in galactose media and spot-testing on galactose plates starting with an equal number of yeast cells as determined by taking the OD$_{600}$. Ten-fold serial dilutions were used in the spot-test. Plates were grown at 3$0^{\circ}C$ for 2-3 days. All murine NDPKs suppressed Bax dependent apoptosis. Futher study will be peformed whether Bax-toxicity inhibition was caused by NDP kinase activity or additional function.n.

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Production and Characterization of vitellogenin monoclonal antibody on the Scorpion fish Sebastiscus marmoratus (쏨뱅이, Sebastiscus marmoratus의 vitellogenin 단클론 항체생산 및 특성에 관한 연구)

  • Kim, Young-Ju;Lim, Yoon-Kyu;Yeo, In-Kyu
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.241-254
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    • 2013
  • In order to establish bio-marker systems for the screening of endocrine-disrupting chemicals contaminated in various environment, Vitellogenin(Vtg) bio-marker have been developed to detect Scorpion fish's(Sebastiscus marmoratus) Vtg. Vtg has been induced by administration of estradiol into S. marmoratus, and purified by gel filtration and ion-exchange chromatography from serum of the fish. After immunization of the purified Vtg into BALB/c mouse, hybridomas secreting anti-Vtg antibodies have been produced. The size of induced Vtg in the serum was about 440 kDa by gel filtration using Sepharose CL-6B. By SDS-PAGE analysis, the main band of Vtg, however, was at 175 kDa, and several minor bands have been detected with the main band. Eight different monoclonal antibodies have been produced from established hybridomas and the antibodies did not cross-react with sera from different species of fishes tested in this study except with that of Sebastes hubbsi. These results suggested that the monoclonal antibody of S28 and S15 can used as capture and tracer antibodies for ELISA and ICG assays. The detection systems developed in this study can be used as Bio-marker assays to check endocrine disrupting activity of various chemicals as well as to detect known endocrine disrupting chemicals contaminated in environment.

Study on the Correlation between the Soil Properties and Albiflorin, Paeoniflorin Contents of Paeonia lactiflora Pall. (작약의 Albiflorin, Paeoniflorin 함량과 토양특성 간의 상관관계 연구)

  • Eo, Hyun Ji;Park, Youngki;Park, Gwang Hun;Kim, Ji-Ah;Kim, Da Som;Kang, Yeongyeong;Kim, Kiyoon;Jang, Jun Hyuk;Kim, Hyun-Jun
    • Korean Journal of Plant Resources
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    • v.34 no.4
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    • pp.384-394
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    • 2021
  • The aim of this study was to investigate the correlation between soil properties and marker compounds contents of Paeonia lactiflora. The methods of determining marker compounds were validated by measuring the linearity, limit of detection (LOD), limit of quantification (LOQ), precision, accuracy and recovery using UPLC analysis. P. lactiflora contained albiflorin at 0.04 ± 0.00 ~ 2.79 ± 0.21%, paeoniflorin at 1.98 ± 0.14 ~ 6.67 ± 0.84%. The root dry weight (RDW) of P. lactiflora was 0.06 ± 0.02 ~ 1.27 ± 0.28 kg. The soil properties analysis such as soil pH, electric conductivity (EC), organic matter (OM), total nitrogen (TN), available phosphate (Avail. P2O5), exchangeable cation and cation exchange capacity (CEC) were performed following standard analysis manual. The results of correlation analysis between soil properties and growth characteristics, available P2O5 was positively coreelated with the RDW of P. lactiflora. On the other hand, the RDW of P. lactiflora showed significantly negative correlation with contents of albiflorin and paeoniflorin. The results of this study was might be help to provide useful information on the establish of standard cultivation by the investigate correlation analysis between growth characteristics and marker compound contents of P. lactiflora.

A Preparative Purification Process for Recombinant Hepatitis B Core Antigen Using Online Capture by Expanded Bed Adsorption Followed by Size-Exclusion Chromatography

  • Ho, Chin Woi;Tan, Wen Siang;Chong, Fui Chin;Ling, Tau Chuan;Tey, Beng Ti
    • Journal of Microbiology and Biotechnology
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    • v.19 no.4
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    • pp.416-423
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    • 2009
  • Hepatitis B core antigen(HBcAg) is an important serological marker used in the diagnosis of hepatitis B virus(HBV) infections. In the current study, a fast and efficient preparative purification protocol for truncated HBcAg from Escherichia coli disruptate was developed. The recombinant HBcAg was first captured by anion exchange expanded bed adsorption chromatography integrated with a cell disruption process. This online capture process has shortened the process time and eliminated the "hold-up" period that may be detrimental to the quality of target protein. The eluted product from the expanded bed adsorption chromatography was subsequently purified using size-exclusion chromatography. The results showed that this novel purification protocol achieved a recovery yield of 45.1% with a product purity of 88.2%, which corresponds to a purification factor of 4.5. The recovered HBcAg is still biologically active as shown by ELISA test.

Development of an Interaction Behaviors Checklist for Early Detection of Autistic Children (자폐아동의 조기 선별을 위한 상호작용행동체크리스트 개발)

  • Im, Sook-Bin
    • Journal of Korean Academy of Nursing
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    • v.35 no.1
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    • pp.5-15
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    • 2005
  • Purpose: This study was conducted to develop a behavioral checklist to predict an autistic disorder and to identify the earliest detecting time. Method: One hundred and fifty eight children including normal, autistic, institutionalized normal, and retarded were assessed using critical interaction behavioral markers from literature review. Data was collected by semi-structured mother-child interaction by videotape recording and analyzed byfactor analysis, Cronbach a, Kappa, $x^2$, and Duncan. Result: Ten behavioral markers were sorted into 2 factors; joint-attention and synchronized behavior. Autistic children were impaired in pretend play, odeclarative pointing, proimperative pointing, gaze-monitoring, referential looking, showing, joint-attention, rhythmical vocal exchange, and synchronized laughing. The sychronized behavior was also a critical marker to predict the autistic disorder. However, it was difficult to differentiate autistic disorder from mental retardation. In addition, the appropriate detecting time was around 18 months after birth. Conclusion: This checklist should be behavior markers to predict autistic disorder and could be useful as educational material at children's clinics, parents class, and for caregivers in the health center. In addition, early detection should lead to treatment being started as soon after 18 months of age as possible.