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민승의,박유진,김동명 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.1
In this work, we attempted a sequential expression of different proteins in a same reaction mixture for cell-free protein synthesis. While a cell-free protein synthesis system of the conventional batch configuration could support translation reactions only for limited time periods and protein synthesis stops within an hour, the translational activity in a completed reaction was recovered by dialyzing the reaction mixture against an appropriate buffer. As a result, when the reaction mixture was re-programmed with second DNA template, the encoded protein was successfully generated by using the ‘recycled’ translational machinery. We expect that the proposed approach will offer a facile route to the preparation of protein pairs for studying the structure and function of interacting proteins.
민승의,김동명 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.1
In this study, we describe the use of antisense oligonucleotides for selective repression of gene expression in a cell-free protein synthesis system. When the selected antisense oligonucleotides were added to a reaction mixture of cell-free protein synthesis that had been programmed with multiple template DNAs, expression of the target genes carrying the complementary sequence were selectively repressed. Moreover, the antisense oligonucleotides were able to be used as a reversible switch to control the expression of target genes by conjugating them with a temperature- responsive polymer. Presented results suggest that expression of recombinant proteins can be modulated by physical stimuli and the possibility of modulating the expression of individual genes in the context of complicated networks.
민승의,김동명 한국공업화학회 2014 한국공업화학회 연구논문 초록집 Vol.2014 No.1
The ability to incorporate unnatural amino acids can expand the scope of protein engineering beyond the present limitations of using 20 canonical amino acids. While cell-based production of unnatural amino acid-containing protein is cumbersome and time and labor-intensive process, herein we propose a rapid and efficient tool for translational incorporation of unnatural amino acids into proteins. The advantages of cell-free protein synthesis include speed and throughput in protein production. In addition, the open nature of cell-free protein synthesis allows direct modifications of the components for protein synthesis. While the simple replacements of amino acids led to mixed synthesis of proteins containing unnatural and natural amino acids, in our experiments to replace tyrosine with 3,4-dihydroxy-L-phenylalanine (L-DOPA), almost exclusive incorporation of unnatural amino acids was achieved by washing the cell extract to remove residual natural amino acids in cell extract. Proposed strategy should be able to provide a facile route to selective incorporation unnatural amino acids in protein structure.
Cell-free synthesis of scFv with enhanced solubility and disulfide bond formation
민승의,김동명 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.1
In this study we attempted to express the scFv antibodies that had been engineered to penetrate into the cells and give signal through split GFP complementation. Conventional cell-based gene expression methods often fail to produce sufficient amount of scFv. In contrast, cell-free protein synthesis system provides a high accessibility to the translational mechanism, thereby providing optimal environments to increase the productivity and solubility of scFv. scFv productivity was enhanced by adding a ubiquitin sequence in front of the ORF of scFv sequence. To increase the solubility, we used molecular chaperon GroEL/ES-enriched extract. To form correct disulfide bonds, we have changed redox conditions of the cell extract and reaction mixture using GSSG/GSH and DsbC. We expect that the established conditions would be applied as a general strategy for producing active scFvs.
브랜드 성격을 기반으로 한 정책 수용도와 부처 신뢰의 선행 요인 탐구
민승,김지원,성용준 한국광고홍보학회 2023 한국광고홍보학보 Vol.25 No.2
While a number of policies are being proposed and implemented to better impact citizens’ lives, the poor level of policy acceptance has been a persistent problem. Accordingly, the increase in government department trust as well as in policy acceptance intention must be considered. Numerous studies, thus, have addressed the importance of individual government departments’ branding and have explored conducted studies in terms of the brand personality (Aaker, 1997) perspective, yet most have explored only a few departments. The present study attempted to expand the existing literature by examining how the policy consumers (e.g., citizens) perceive each department’s brand personality and how such dimensions of departments’ brand personality affect trust and policy acceptance. Results showed that respondents recognized the ‘authoritative’ personality dimension as the highest for most of the departments, and the correlations among the eight personality dimensions, trust, and policy acceptance intention were significantly high. In addition, the brand personality dimensions that significantly affects policy acceptance intention mediated by department trust were found to be four dimensions: sincerity, competence, friendliness, and ethicality. These findings provide theoretical implications in expanding the departments’ brand personality literatures and practical implications in providing guidelines for more effective policy implementation to increase policy acceptance. 국민의 삶에 긍정적인 영향을 미치기 위해 다양한 정책들이 제시 및 이행되고 있지만, 정책이 잘 수행되지 못하는 문제가 꾸준히 제기되고 있으며, 이를 위해 정책 대상자(국민)의 정책 수용 의도와 그의 선행 요인으로 부처에 대한 신뢰를 높이는 것이 중요하다. 이에 따라 선행 연구에서는 부처 브랜딩의 필요성을 제시하며 다양한 관점에서 브랜드 성격을 기반으로 탐구하였으나, 대부분 몇 가지의 부처만을 대상으로 진행되었다. 따라서 본 연구에서는 부처 브랜드 성격을 기반으로 부처에 대한 정책 대상자(국민)의 인식이 어떻게 이루어져 있는지, 그리고 이러한 부처 브랜드 성격 측면이 부처 신뢰와 정책 수용도에 어떠한 영향을 미치는지를 살펴봄으로써 기존 연구를 확장하고자 하였다. 설문조사 결과 사람들은 대부분 부처에 대해 8개의 브랜드 성격 요인 중 ‘권위적임’ 성격 측면이 가장 높다고 인식하였으며, 8개의 성격 요인과 부처 신뢰, 정책 수용도 간의 상관관계가 매우 높게 나타났다. 또한, 부처 신뢰를 통해 정책 수용도에 영향을 미치는 부처 브랜드 성격 요인은 ‘진정성’, ‘유능함’, ‘우호적임’, ‘윤리적임’의 4가지 성격 측면으로 나타났다. 이러한 연구 결과는 기존의 공공 맥락에서 지속적으로 탐구되었던 부처 브랜드 성격 인식을 확장하여 살펴보았다는 점에서 이론적 의의가 있으며, 정책 집행하는 과정에서 정책 수용도를 높이는 데 보다 효과적인 정책 이행을 위한 가이드라인을 제시한다는 점에서 실무적 의의가 있다.