• Title/Summary/Keyword: Fusion system

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JSAP1 Interacts with Kinesin Light Chain 1 through Conserved Binding Segments (JSAP1과 Kinesin Light Chain 1의 결합 및 결합부위 규명)

  • Kim, Sang-Jin;Lee, Chul-Hee;Park, Hye-Young;Yea, Sung-Su;Jang, Won-Hee;Lee, Sang-Kyeong;Park, Yeong-Hong;Cha, Ok-Soo;Moon, Il-Soo;Seog, Dae-Hyun
    • Journal of Life Science
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    • v.17 no.7 s.87
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    • pp.889-895
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    • 2007
  • A conventional kinesin, KIF5/kinesin-I, is composed of two kinesin heavy chains (KHCs) and two kinesin light chains (KLCs) and binds directly to microtubules. KIF5 motor mediates the transport of various membranous organelles, but the mechanism how they recognize and bind to a specific cargo has not yet been completely elucidated. Here, we used the yeast two-hybrid system to identify the neuronal protein(s) that interacts with the tetratricopeptide repeats (TRP) of KLCI and found a specific interaction with JNK/stress-activated protein kinase-associated protein 1 (JSAP1/JIPP3). The yeast two-hybrid assay demonstrated that the TRP 1,2 domain-containing region of KLCI mediated binding to the leucine zipper domain of JSAP1. JSAP1 also bound to the TRP region of lac2 but not to neuronal KIF5A, KIF5C and ubiquitous KIF5B in the yeast two-hybrid assay. In addition, these proteins showed specific interactions in the GST pull-down assay and by co-immunoprecipitation. KLCI and KIF5B interacted with GST-ISAP1 fusion proteins, but not with GST alone. An antibody to JSAPI specifically co-immunoprecipitated KIF5s associated with JSAP1 from mouse brain extracts. These results suggest that JSAP1, as KLC1 receptor, is involved in the KIF5 mediated transport.

Purification of Human HtrA1 Expressed in E. coli and Characterization of Its Serine Protease Activity (E. coli에서 발현된 human HtrA1 단백질의 정제와 HtrA1의 serine protease 활성 조건에 관한 연구)

  • Kim, Kyung-Hee;Kim, Sang-Soo;Kim, Goo-Young;Rhim, Hyang-Shuk
    • Journal of Life Science
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    • v.16 no.7 s.80
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    • pp.1133-1140
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    • 2006
  • Human HtrA1 (High temperature requirement protein A1) is a homologue of the E. coli periplasmic serine protease HtrA. A recent study has demonstrated that HtrA1 is a serine protease involved in processing of insulin like growth factor binding protein (ICFBP), indicating that it serves as an important regulator of IGF activity. Additionally, several lines of evidence suggest a striking correlation between proteolytic activity of HtrA1 serine protease and the pathogenesis of several diseases; however, physiological roles of HtrA1 remain to be elucidated. We used the pGEX bacterial expression system to develop a simple and rapid method for purifying HtrA1, and the recombinant HtrA1 protein was utilized to investigate the optimal conditions in executing its proteolytic activity. The proteolytically active HtrA1 was purified to approximately 85% purity, although the yield of the recombinant HtrA1 protein was slightly low $460{\mu}g$ for 1 liter E. coli culture). Using in vitro endoproteolytic cleavage assay, we identified that the HtrA1 serine protease activity was dependent on the enzyme concentration and the incubation time and that the best reaction temperature was $42^{\circ}C$ instead of $37^{\circ}C$. We arbitrary defined one unit of proteolytic activity of the HtrA1 serine protease as 200nM of HtrA1 that cleaves half of $5{\mu}M\;of\;{\beta}-casein$ during 3 hr incubation at $37^{\circ}C$. Our study provides a method for generating useful reagents to investigate the molecular mechanisms by which HtrA1 serine protease activity contributes in regulating its physiological function and to identify natural substrates of HtrA1.

Performance Characteristics of PM10 and PM2.5 Samplers with an Advanced Chamber System (챔버 기술 개발을 통한 PM10과 PM2.5 시료채취기의 수행 특성)

  • Kim, Do-Hyeon;Kim, Seon-Hong;Kim, Ji-Hoon;Cho, Seung-Yeon;Park, Ju-Myon
    • Journal of Korean Society of Environmental Engineers
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    • v.32 no.8
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    • pp.739-746
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    • 2010
  • The purposes of this study are 1) to develop an advanced chamber system within ${\pm}10%$ of air velocity at the particulate matter (PM) collection area, 2) to research theoretical characteristics of PM10 and PM2.5 samplers, 3) to assess the performance characteristics of PM10 and PM2.5 samplers through chamber experiments. The total six one-hour experiments were conducted using the cornstarch with an mass median aerodynamic diameter (MMAD) of $20\;{\mu}m$ and an geometric standard deviation of 2.0 at the two different air velocity conditions of 0.67 m/s and 2.15 m/s in the chamber. The aerosol samplers used in the present study are one APM PM10 and one PM2.5 samplers accordance with the US federal reference methods and specially designed three mini-volume aerosol samplers (two for PM10 and one for PM2.5). The overall results indicate that PM10 and PM2.5 mini-volume samplers need correction factors of 0.25 and 0.39 respectively when APM PM samplers considered as reference samplers and there is significant difference between two mini-volume aerosol samplers when a two-way analysis of variance is tested using the measured PM10 mass concentrations. The PM10 and PM2.5 samplers with the cutpoints and slopes (PM10: $10{\pm}0.5\;{\mu}m$ and $1.5{\pm}0.1$, PM2.5: $2.5{\pm}0.2\;{\mu}m$ and $1.3{\pm}0.03$) theoretically collect the ranges of 86~114% and 64~152% considering the cornstarch characteristics used in this research. Furthermore, the calculated mass concentrations of PM samplers are higher than the ideal mass concentrations when the airborne MMADs for the cornstarch used are smaller than the cutpoints of PM samplers and the PM samplers collected less PM in another case. The chamber experiment also showed that PM10 and PM2.5 samplers had the bigger collection ranges of 37~158% and 55~149% than the theocratical calculated mass concentration ranges and the relatively similar mass concentration ranges were measured at the air velocity of 2.15 m/s comparing with the 0.67 m/s.

A Novel Glycine-Rich Region in Sox4 is a Target for the Proteolytic Cleavage in E. coli (전사활성 인자인 Sox4의 단백질 분해효소에 의한 표적 부위에 관한 연구)

  • 허은혜;최주연;장경희;김인경;임향숙
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.153-161
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    • 2002
  • Sox4, a transcription factor, consists of three functional domains: an HMG-box domain as a DNA binding domain, serine rich region as a transactivation domain and glycine rich region (GRR), an unknown functional domain. Although Sox4 is known to be functionally involved in heart, B-cell and reproductive system development, its physiological function remains to be elucidated. We used pGEX expression system to develop a simple and rapid method for purifying Sox4 protein in suitable forms for biochemical studies of their functions. Unexpectedly, we observed that full-length Sox4 appears to be protease-sensitive during expression and purification in E. coli. To map the protease-sensitive site in Sox4, we generated various constructs with each of functional domains of Sox4 and purified as the GST-Sox4 fusion proteins using glutathione beads. We found that the specific cleavage site for the proteolytic enzyme, which exists in E. coli, is localized within the novel GRR of Sox4. Our study suggest that the GRR of Sox4 may a target for the cellular protease action and this cleavage in the GRR may be involved in regulating physiological function of Sox4. Additionally, our study may provide a useful method for investigating the proteolytic cleavage of the target molecule in E. coli.

Inhibitory Effort of the N-terminal GST on the Tautomerase Activity of Macrophage Migration Inhibitory Factor (GST 융합 시스템에서 나타나는 macrophage migration inhibitory factor의 tautomerase 활성 저해에 관한 연구)

  • Kim Sang-Soo;Kim Kyung-Hee;Park Hyo-Jin;Hur Eun-hye;Rhim Hyangshuk
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.961-967
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    • 2005
  • Macrophage migration inhibitory fartor (MIF), known as a cytokine, is a multifunctional protein that is ubiquitously expressed in a variety of cells and tissues; however, enzymatic function of MIF still remains elusive in cells. In this study, we assessed details of the tautomerase activity of MIF. We established rapid purification condition for MIF by using pGEX system and compared the L-dopachrome tautomerase activity of GST-MIF, tMIF, and MIF. The results show that GST (glutathione S-transferase)-epitope tag or N-terminal amino acids flanking the essential $P^{2}$ almost completely abrogated L-dopachrome tautomerase activity of MIF. Subsequently, to determine whether the N-terminal tags have effects on oligomerization of MIF, protein cross-linking products were analyzed on $15\%$ SDS-PACE. The result demonstrates that N-terminal tags are dispensable for the formation of MIF's homooligomers. Thus, the results imply that exposure of If containing hydrophobic pocket in the active site is critical for L-dopachrome tautomerase activity of MIF. In addition, our study suggest that the MIF's tautomerase activity might be influenced by interacting with cellular partners.

Exploring the 4th Industrial Revolution Technology from the Landscape Industry Perspective (조경산업 관점에서 4차 산업혁명 기술의 탐색)

  • Choi, Ja-Ho;Suh, Joo-Hwan
    • Journal of the Korean Institute of Landscape Architecture
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    • v.47 no.2
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    • pp.59-75
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    • 2019
  • This study was carried out to explore the 4th Industrial Revolution technology from the perspective of the landscape industry to provide the basic data necessary to increase the virtuous circle value. The 4th Industrial Revolution, the characteristics of the landscape industry and urban regeneration were considered and the methodology was established and studied including the technical classification system suitable for systematic research, which was selected as a framework. First, the 4th Industrial Revolution technology based on digital data was selected, which could be utilized to increase the value of the virtuous circle for the landscape industry. From 'Element Technology Level', and 'Core Technology' such as the Internet of Things, Cloud Computing, Big Data, Artificial Intelligence, Robot, 'Peripheral Technology', Virtual or Augmented Reality, Drones, 3D 4D Printing, and 3D Scanning were highlighted as the 4th Industrial Revolution technology. It has been shown that it is possible to increase the value of the virtuous circle when applied at the 'Trend Level', in particular to the landscape industry. The 'System Level' was analyzed as a general-purpose technology, and based on the platform, the level of element technology(computers, and smart devices) was systematically interconnected, and illuminated with the 4th Industrial Revolution technology based on digital data. The application of the 'Trend Level' specific to the landscape industry has been shown to be an effective technology for increasing the virtuous circle values. It is possible to realize all synergistic effects and implementation of the proposed method at the trend level applying the element technology level. Smart gardens, smart parks, etc. have been analyzed to the level they should pursue. It was judged that Smart City, Smart Home, Smart Farm, and Precision Agriculture, Smart Tourism, and Smart Health Care could be highly linked through the collaboration among technologies in adjacent areas at the Trend Level. Additionally, various utilization measures of related technology applied at the Trend Level were highlighted in the process of urban regeneration, public service space creation, maintenance, and public service. In other words, with the realization of ubiquitous computing, Hyper-Connectivity, Hyper-Reality, Hyper-Intelligence, and Hyper-Convergence were proposed, reflecting the basic characteristics of digital technology in the landscape industry can be achieved. It was analyzed that the landscaping industry was effectively accommodating and coordinating with the needs of new characters, education and consulting, as well as existing tasks, even when participating in urban regeneration projects. In particular, it has been shown that the overall landscapig area is effective in increasing the virtuous circle value when it systems the related technology at the trend level by linking maintenance with strategic bridgehead. This is because the industrial structure is effective in distributing data and information produced from various channels. Subsequent research, such as demonstrating the fusion of the 4th Industrial Revolution technology based on the use of digital data in creation, maintenance, and service of actual landscape space is necessary.

The Causal Relation between Win-Win Growth Strategies of Small and Medium-Sized Businesses and Corporate Performance (중소기업의 동반성장 전략과 기업성과의 인과 관계)

  • Ban, Won Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.19 no.12
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    • pp.552-560
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    • 2018
  • Since 1960's, the large conglomerates of South Korea have grown due to the corporate-centered, fast-paced growth drive, while the small and medium-sized businesses supported the country's economy as the subordinate structure of these conglomerates. Due to the globalization of the business environments, the focus of competition shifted from competitions between individual companies to one between networks of companies. Therefore, more emphasis is now put on the capabilities of the cooperation networks between companies rather than the capabilities of individual companies. Therefore, in this study, the author examined the influence of the win-win growth strategy elements through cooperation with small and medium-sized businesses upon corporate performance. This study was conducted with the workers of small and medium-sized businesses that have previous cooperation experiences with South Korean conglomerates over the period from March 2 to May 17, 2018. For this, a total of 515 questionnaires were retrieves to obtain the data for analysis. The analysis was conducted using SPSS 22.0 and AMOS 18.0. The analytical processes that were taken included exploratory factor analysis, confirmatory factor analysis, confidence analysis, correlation analysis, and structural equation analysis model. The results of the analysis showed that, first of all, the win-win growth strategy factors that affected the strategic performance, which is a part of cooperate performance were, respectively, harmonization with the goals, production technical support, and quality system. Second, the win-win growth strategy factors that affected the financial performance, which is a part of corporate performance, turned out to be harmonization with the goals, quality system, and incentive. With the results of this study, it was shown that the elements such as harmonization with the goals, production technical support, quality systems, and incentives were key infrastructural factors that affected the corporate performance directly. On the other hand, its implication is that informative or knowledge-related factors, such as joint knowledge creation, do not have their own added values, while they are not too much likely to affect corporate performances for the moment.

The Policy of Win-Win Growth between Large and Small Enterprises : A South Korean Model (한국형 동반성장 정책의 방향과 과제)

  • Lee, Jang-Woo
    • Korean small business review
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    • v.33 no.4
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    • pp.77-93
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    • 2011
  • Since 2000, the employment rate of small and medium enterprises (SMEs) has dwindled while the creation of new jobs and the emergence of healthy SMEs have been stagnant. The fundamental reason for these symptoms is that the economic structure is disadvantageous to SMEs. In particular, the greater gap between SMEs and large enterprises has resulted in polarization, and the resulting imbalance has become the largest obstacle to improving SMEs' competitiveness. For example, the total productivity has continued to drop, and the average productivity of SMEs is now merely 30% of that of large enterprises, and the average wage of SMEs' employees is only 53% of that of large enterprises. Along with polarization, rapid industrialization has also caused anti-enterprise consensus, the collapse of the middle class, hostility towards establishments, and other aftereffects. The general consensus is that unless these problems are solved, South Korea will not become an advanced country. Especially, South Korea is now facing issues that need urgent measures, such as the decline of its economic growth, the worsening distribution of profits, and the increased external volatility. Recognizing such negative trends, the MB administration proposed a win-win growth policy and recently introduced a new national value called "ecosystemic development." As the terms in such policy agenda are similar, however, the conceptual differences among such terms must first be fully understood. Therefore, in this study, the concepts of win-win growth policy and ecosystemic development, and the need for them, were surveyed, and their differences from and similarities with other policy concepts like win-win cooperation and symbiotic development were examined. Based on the results of the survey and examination, the study introduced a South Korean model of win-win growth, targeting the promotion of a sound balance between large enterprises and SMEs and an innovative ecosystem, and finally, proposing future policy tasks. Win-win growth is not an academic term but a policy term. Thus, it is less advisable to give a theoretical definition of it than to understand its concept based on its objective and method as a policy. The core of the MB administration's win-win growth policy is the creation of a partnership between key economic subjects such as large enterprises and SMEs based on each subject's differentiated capacity, and such economic subjects' joint promotion of growth opportunities. Its objective is to contribute to the establishment of an advanced capitalistic system by securing the sustainability of the South Korean economy. Such win-win growth policy includes three core concepts. The first concept, ecosystem, is that win-win growth should be understood from the viewpoint of an industrial ecosystem and should be pursued by overcoming the issues of specific enterprises. An enterprise is not an independent entity but a social entity, meaning it exists in relationship with the society (Drucker, 2011). The second concept, balance, points to the fact that an effort should be made to establish a systemic and social infrastructure for a healthy balance in the industry. The social system and infrastructure should be established in such a way as to create a balance between short- term needs and long-term sustainability, between freedom and responsibility, and between profitability and social obligations. Finally, the third concept is the behavioral change of economic entities. The win-win growth policy is not merely about simple transactional relationships or determining reasonable prices but more about the need for a behavior change on the part of economic entities, without which the objectives of the policy cannot be achieved. Various advanced countries have developed different win-win growth models based on their respective cultures and economic-development stages. Japan, whose culture is characterized by a relatively high level of group-centered trust, has developed a productivity improvement model based on such culture, whereas the U.S., which has a highly developed system of market capitalism, has developed a system that instigates or promotes market-oriented technological innovation. Unlike Japan or the U.S., Europe, a late starter, has not fully developed a trust-based culture or market capitalism and thus often uses a policy-led model based on which the government leads the improvement of productivity and promotes technological innovation. By modeling successful cases from these advanced countries, South Korea can establish its unique win-win growth system. For this, it needs to determine the method and tasks that suit its circumstances by examining the prerequisites for its success as well as the strengths and weaknesses of each advanced country. This paper proposes a South Korean model of win-win growth, whose objective is to upgrade the country's low-trust-level-based industrial structure, in which large enterprises and SMEs depend only on independent survival strategies, to a high-trust-level-based social ecosystem, in which large enterprises and SMEs develop a cooperative relationship as partners. Based on this objective, the model proposes the establishment of a sound balance of systems and infrastructure between large enterprises and SMEs, and to form a crenovative social ecosystem. The South Korean model of win-win growth consists of three axes: utilization of the South Koreans' potential, which creates community-oriented energy; fusion-style improvement of various control and self-regulated systems for establishing a high-trust-level-oriented social infrastructure; and behavioral change on the part of enterprises in terms of putting an end to their unfair business activities and promoting future-oriented cooperative relationships. This system will establish a dynamic industrial ecosystem that will generate creative energy and will thus contribute to the realization of a sustainable economy in the 21st century. The South Korean model of win-win growth should pursue community-based self-regulation, which promotes the power of efficiency and competition that is fundamentally being pursued by capitalism while at the same time seeking the value of society and community. Already existing in Korea's traditional roots, such objectives have become the bases of the Shinbaram culture, characterized by the South Koreans' spontaneity, creativity, and optimism. In the process of a community's gradual improvement of its rules and procedures, the trust among the community members increases, and the "social capital" that guarantees the successful control of shared resources can be established (Ostrom, 2010). This basic ideal can help reduce the gap between large enterprises and SMEs, alleviating the South Koreans' victim mentality in the face of competition and the open-door policy, and creating crenovative corporate competitiveness. The win-win growth policy emerged for the purpose of addressing the polarization and imbalance structure resulting from the evolution of 21st-century capitalism. It simultaneously pursues efficiency and fairness on one hand and economic and community values on the other, and aims to foster efficient interaction between the market and the government. This policy, however, is also evolving. The win-win growth policy can be considered an extension of the win-win cooperation that the past 'Participatory Government' promoted at the enterprise management level to the level of systems and culture. Also, the ecosystemic development agendum that has recently emerged is a further extension that has been presented as a national ideal of "a new development model that promotes the co-advancement of environmental conservation, growth, economic development, social integration, and national and individual development."

Effects of Encapsulation Layer on Center Crack and Fracture of Thin Silicon Chip using Numerical Analysis (봉지막이 박형 실리콘 칩의 파괴에 미치는 영향에 대한 수치해석 연구)

  • Choa, Sung-Hoon;Jang, Young-Moon;Lee, Haeng-Soo
    • Journal of the Microelectronics and Packaging Society
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    • v.25 no.1
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    • pp.1-10
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    • 2018
  • Recently, there has been rapid development in the field of flexible electronic devices, such as organic light emitting diodes (OLEDs), organic solar cells and flexible sensors. Encapsulation process is added to protect the flexible electronic devices from exposure to oxygen and moisture in the air. Using numerical simulation, we investigated the effects of the encapsulation layer on mechanical stability of the silicon chip, especially the fracture performance of center crack in multi-layer package for various loading condition. The multi-layer package is categorized in two type - a wide chip model in which the chip has a large width and encapsulation layer covers only the chip, and a narrow chip model in which the chip covers both the substrate and the chip with smaller width than the substrate. In the wide chip model where the external load acts directly on the chip, the encapsulation layer with high stiffness enhanced the crack resistance of the film chip as the thickness of the encapsulation layer increased regardless of loading conditions. In contrast, the encapsulation layer with high stiffness reduced the crack resistance of the film chip in the narrow chip model for the case of external tensile strain loading. This is because the external load is transferred to the chip through the encapsulation layer and the small load acts on the chip for the weak encapsulation layer in the narrow chip model. When the bending moment acts on the narrow model, thin encapsulation layer and thick encapsulation layer show the opposite results since the neutral axis is moving toward the chip with a crack and load acting on chip decreases consequently as the thickness of encapsulation layer increases. The present study is expected to provide practical design guidance to enhance the durability and fracture performance of the silicon chip in the multilayer package with encapsulation layer.

Characterization of SID2 that is required for the production of salicylic acid by using β-GLUCURONIDASE and LUCIFERASE reporter system in Arabidoposis (리포트 시스템을 이용한 살리실산 생합성 유전자 SID2의 발현 해석)

  • Hong, Mi-Ju;Cheong, Mi-Sun;Lee, Ji-Young;Kim, Hun;Jeong, Jae-Cheol;Shen, Mingzhe;Ali, Zahir;Park, Bo-Kyung;Choi, Won-Kyun;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.169-176
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    • 2008
  • Salicylic acid(SA) is a phytohormone that is related to plant defense mechanism. The SA accumulation is triggered by abiotic and biotic stresses. SA acts as a signal molecular compound mediating systemic acquired resistance and hypersensitive response in plant. Although the role of SA has been studied extensively, an understanding of the SA regulatory mechanism is still lacking in plants. In order to comprehend SA regulatory mechanism, we have been transformed with a SID2 promoter:GUS::LUC fusion construct into siz1-2 mutant and wild plant(Col-0). SIZ1 encodes SUMO E3 ligase and negatively regulates SA accumulation in plants. SID2(SALICYLIC ACID INDUCTION DEFICIENT2) is a crucial enzyme of SA biosynthesis. The Arabidopsis SID2 gene encodes isochorismate synthase(ICS) that controls SA level by conversion of chorismate to isochorismate. We compared the regulation of SID2 in wild-type and siz1-2 transgenic plants that express SID2 promoter:GUS::LUC constructs respectively. The expressions of $\beta$-GLUCURONIDASE and LUCIFERASE were higher in siz 1-2 transgenic plant without any stress treatment. SID2 promoter:GUS::LUC/siz1-2 transgenic plant will be used as a starting material for isolation of siz1-2 suppressor mutants and genes involved in SA-mediated stress signaling pathway.