• Microbiology and Biotechnology Letters
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• v.24 no.5
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• pp.604-612
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• 1996

To improve the fermentation characteristics of the haploid starch-fermenting recombinant yeast strain K114/YIpMS$\Delta$R(LEU2/URA3) secreting both $\alpha$-amylase and glucoamylase was rare-mated with polyploid industrial yeast Saccharomyces sp. K35. The K35 strain had good fermentation-characteristics such as ethanol-tolerance, high temperature and sugar-tolerance, and high fermentation rate. Among the resulting 66 hybrids, the best strain RH51 was selected. The RH51 exhibited amylolytic activity of K114/YIpMS$\Delta$R(LEU2/URA3) as well as ethanol and sugar tolerance of K35. The optimum temperature of hybrid RH51 for starch fermentation was 34$\circ$C which was same as that of K35 but different from that (30$\circ$C) of K114/YIpMS$\Delta$R(LEU2/URA3). The optimum pH was 5.0. The optimum size of inoculum was 2% as the pellet (w/v) of yeast cells. The hybrid strain RH51 produced 7.0% ethanol (w/v) from 20% (w/v) soluble starch while K35 formed almost no ethanol, 0.3% (w/v). RH51 strain produced 7.5% (w/v) ethanol after 8 days in a 2.5 l fermenter containing 800 ml of 20% (w/v) soluble starch. The residual starch content in the fermentation medium was 1.68% (w/v), and therefore almost all the starch was fermented completely.

• Korean Journal of Microbiology
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• v.41 no.4
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• pp.239-245
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• 2005

Transformation-Associated Recombination (TAR) cloning technique allows selective isolation of chromosomal regions and genes from complex genomes. The procedure requires knowledge of relatively small genomic sequences that reside adjacent to the chromosomal region of interest. This technique involves homologous recombination during yeast spheroplast transformation between genomic DNA and a TAR vector that has 5' and 3' gene targeting sequences. In this study, we examined the effect of non-homologous spacing sequence in target hooks on homologous recombination using a plasmid model system. The efficiency of homologous recombination between the modified his3-TRP1-his3 fragments and HlS3 gene on plasmid were analyzed by the characterization of $Ura^+$ transformants. The numbers of $Ura^+$ transformant showed same level when seven different modified his3-TRP1-his3 fragments were used. But the percentage of positive recombinants. $Trp^+His^-$, dramatically decreased when used the modified his3-TRP1-his3 fragments contained incorrect spacing of nonhomologous region. As a result, we suggest that incorrect spacing inhibits the homologous recombination between target hook and substrate DNA. Therefore, we should consider the correct spacing in target hook when the target hook are used for cloning of orthologue gene.

• International conference on construction engineering and project management
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• 2020.12a
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• pp.267-277
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• 2020

The present research evaluates the public engagement approach in various project stages. Hong Kong had long been criticized as top-down and executive-led jurisdiction by overlooking the importance of cultural heritage and public concerns on public projects. It was suggested to the government to engage public and provide sufficient public consultation. Thereby, the government announced a series of revitalization and conversion measures in Policy Address in 2008. To carry out the measures, there were voices, because of diverse and sometimes conflicting interests, over the effect of revitalization project. On the other hand, studies reveal that there are benefits of revitalization and enhancement of public engagement approach. In pursuit of the subject, the present research aims at studying the Central Market as a case study pilot project. In October 2009, the Policy Address announced that the Central Market be revitalized. Tasked to implement the project, Urban Renewal Authority (URA) continued to adopt the people-oriented approach as the guiding principle in its core business and in heritage preservation and revitalization so as to create a sustainable development. Between government and the public, URA acts as a bridge for communication. As between URA and the public, URA conducted public consultation, set up an ad-hoc committee in January 2016 to be in charge of the project and will continuously inform the government and public with the updates and project progress. The main objectives of the present research are to assess the advantages and challenges in different stages of public engagement approach, to evaluate the engagement system, to give a comprehensive view for participation of stakeholders, and to find out effective strategies to enhance civil engagement. Research was achieved through interviews to key persons in the project, questionnaire that was distributed to community and experts in the field. Case Study of the Central Market was studied and investigated using different sources, such as newspapers, journals, etc, to evaluate the degree of public engagement in the project. Both detailed qualitative methodological approaches of interview, questionnaire, and case study, act as a synergy to demonstrate the research objectives and provide the comprehensive perceptions on the revitalization project. The results show that public participants in revitalized project have generated considerable value enhancements to social-cultural, environmental, political and economic aspect. This study provides valuable insights that the public participation can make positive contribution to sustainability in the city. The findings indicate that no any single system is flawless therefore seeking both public opinions and professional advices are also important as a comprehensive approach to achieve higher sustainability of the building.

• Bulletin of the Korean Chemical Society
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• v.24 no.4
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• pp.524-526
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• 2003

• Proceedings of the Korea Water Resources Association Conference
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• 2005.09b
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• pp.982-983
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• 2005

• Bulletin of the Korean Chemical Society
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• v.28 no.8
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• pp.1429-1432
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• 2007

• Journal of the Korean Society for Aviation and Aeronautics
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• v.22 no.1
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• pp.22-31
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• 2014

Military UAV(Unmanned Aerial Vehicle) operated as a RC(Remotely Control) model level within the limit of the military special use airspace until now. However, the high and medium altitude of URA(Unmanned Reconnaissance Aircraft) which the ROKAF have been trying to import recently is at the UAV level and needs the criteria for the classified airspace flights. The required flight criteria includes operator location, mission operation limit, equipment, etc., which are the principle and standard applied based on the airspace use for UAV. Also, the general flight rules, visual flight rules, instrument flight rules are required in order to have to be applied to the actual flight. Besides, an appliance regulation needs to be arranged regarding two-way communication, ATC and communication issue, airspace and area in-flight between UAS( Unmanned Aircraft System) users. An operation of the UAV in the air significantly requires the guarantee of the aircraft's capacity, and also the standardized flight criteria. A safe and smooth use is ensured only if this criteria is applied and understood by the entire airspace users. For the purpose, a standardized military UAV flight operations criteria that is to be applied for each airspace by UAV is to be prepared through analysis of the present state, a legend UAV system, and a special character analysis.

• Journal of Microbiology and Biotechnology
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• v.19 no.3
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• pp.271-276
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• 2009

Functional gene studies in the cultivated white button mushroom Agaricus bisporus have been constrained by the absence of effective gene-silencing tools. Using two endogenous genes from A. bisporus, we have tested the utility of dsRNA hairpin constructs to mediate downregulation of specific genes. Hairpin constructs for genes encoding orotidine 5'-monophosphate decarboxylase (URA3) and carboxin resistance (CBX) were introduced into A. bisporus using Agrobacteriummediated transfection. Although predicted changes in phenotype were not observed in vitro, quantitative-PCR analyses indicated unambiguously that transcripts in several transformants were substantially reduced compared with the non-transformed controls. Interestingly, some hairpin transformants exhibited increased transcription of target genes. Our observations show that hairpin transgenic sequences can mediate downregulation of A. bisporus endogenous genes and that the technology has the potential to expedite functional genomics of the mushroom.

• Microbiology and Biotechnology Letters
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• v.23 no.5
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• pp.536-543
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• 1995

Multicopy integration vector is a very useful vector system in that they can be integrated into chromosomal DNA in several copies and stably maintained under non-selective conditions. To develop a multicopy integration vector system in the yeast Yarrowia lipolytica, P-type ribosomal DNA was cloned from Y lipolytica. A HindIII-BglII fragment of the cloned rDNA and a promoterless URA3 gene were inserted into pGEM1, generating multicopy integration vectors, pMIYL-1 and pMIYL-2. The rDNA fragment is for targeted homologous recombination between the vector and the chromosomal DNA of Y. lipolytica, and the promoterless URA3 gene is a defective selection marker for inducing multicopy integration. pMIYL-1 and pMIYL-2 have an unique restriction enzyme site, KpnI, and two unique restriction enzyme sites, KpnI and EcoRI, repectively, which can be used for targeting of the vectors into the rDNA of Y. lipolytica chromosomal DNA. After transformation of the vectors into Y. lipolytica, copy number and stability were analyzed by Southern hybridization. The vectors were found to be present in less than 5 copies per cell and were stably maintained during growth in non-selective media.

• Journal of Microbiology
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• v.40 no.3
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• pp.245-247
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• 2002

A genomic DNA library of the fungus Trametes versicolor was constructed in a yeast integration vector which contains the URA3 gene of the budding yeast Saccharomyces cerevisiae and the gene responsible for hygromycin B resistance, and fragments acting as autonomously replicating sequences (ARSes) in the budding yeast were identified from the genomic DNA library. Sixteen recombinant plasmids from the library transformed the budding yeast Saccharomyces cerevisiae to Ura+ at high frequencies. They were maintained stably under selective conditions, but were gradually lost from yeast cells at different rates under nonselective conditions, indicating that they contain eukaryotic origins of DNA replication and exist as extrachromosomal plasmids. Base sequences of four ARS DNAs among the 16 cloned fragments revealed that all or the four contain at least one 11 bp ［(A/T)TTTA(T/C)(A/G)TTT(A/T)］consensus sequence of the budding yeast ARS.