• Title/Summary/Keyword: e-NIE

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Monoclonal antibody production for CP4 EPSPS detection assays (CP4 EPSPS 검출을 위한 단클론 항체 생산)

  • A-Mi Yoon;Il Ryong Kim;Wonkyun Choi
    • Korean Journal of Environmental Biology
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    • v.39 no.4
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    • pp.445-451
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    • 2021
  • In this study, we described the production of an antibody to living modified organisms (LMOs) containing the gene encoding for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) from Agrobacterium tumefaciens strain CP4 EPSPS provides resistance to the herbicide glyphosate (N- (phosphonomethyl) glycine). These LMOs were approved and have recently been used in the feed, food production, and processing industries in South Korea. Highly efficient monoclonal antibody (mAb) production is crucial for developing assays that enable the proper detection and quantification of the CP4 EPSPS protein in LMOs. This study describes the purification and characterization of recombinant CP4 EPSPS protein in E. coli BL21 (DE3) based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrixassisted laser desorption/ionization time-of-flight mass spectrometry. The production of mAbs was undertaken based on the standard operating procedure of Abclon, Inc.(South Korea), and the purity of the mAbs was assessed using SDS-PAGE. The following five mAb clones were produced: 2F2, 4B9, 6C11, 10A9, and 10G9. To verify the efficiency and specificity of the five developed mAbs, we performed Western blotting analysis using the LM (living modified) cotton crude extracts. All mAbs could detect the CP4 EPSPS protein in the LM cotton traits MON1445 and MON88913 with high specificity, but not in any other LM cottons or non-LM cottons. These data indicate that these five mAbs to CP4 EPSPS could be successfully used for the further development of antibody-based detection methods to target CP4 EPSPS protein in LMOs.

Changes in Photosynthetic Performance and Water Relation Parameters in the Seedlings of Korean Dendropanax Subjected to Drought Stress (건조 스트레스에 따른 황칠나무 유묘의 광합성과 수분특성인자 변화)

  • Lee, Kyeong Cheol
    • Korean Journal of Medicinal Crop Science
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    • v.26 no.2
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    • pp.181-187
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    • 2018
  • Background: This study aimed to investigate out the influence of drought stress on the physiological responses of Dendropanax morbifera seedlings. Methods and Results: Drought stress was induced by discontinuing water supply for 30 days. Under drought stress, photosynthetic activity was significantly reduced with decreasing soil water content (SWC), as revealed by the parameters such as Fv/Fm, maximum photosynthetic rate ($P_{N\;max}$), stomatal conductance ($g_s$), stomatal transpiration rate (E), and intercellular $CO_2$ concentration (Ci). However, water use efficiency (WUE) was increased by 2.5 times because of the decrease in $g_s$ to reduce transpiration. Particularly, E and $g_s$ were remarkably decreased when water was withheld for 21 days at 6.2% of SWC. Dendropanax morbifera leaves showed osmotic adjustment of -0.30 MPa at full turgor and -0.13 MPa at zero turgor. In contrast, the maximum bulk modulus of elasticity ($E_{max}$) did not change significantly. Thus, Dendropanax morbifera seedlings could tolerate drought stress via osmotic adjustment. Conclusions: Drought avoidance mechanisms of D. morbifera involve reduction in water loss from plants, through the control of stomatal transpiration, and reduction in cellular osmotic potential. Notably photosynthetic activity was remarkably reduced, to approximately 6% of the SWC.

Development of a Screening System for Drugs Against Human Papillomavirus-Associated Cervical Cancer: Based On E7-Rb Binding

  • Cho, Young-Sik;Cho, Cheong-Weon;Kang, Jeong-Woo;Cho, Min-Chul;Lee, Kyung-Ae;Shim, Jung-Hyun;Kwon, Our-Han;Choe, Yong-Kyung;Park, Sue-Nie;Yoon, Do-Young
    • BMB Reports
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    • v.34 no.1
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    • pp.80-84
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    • 2001
  • The human papillomavirus E7 protein can form a specific complex with a retinoblastoma tumor suppressor gene product (p105-Rb) that results in the release of the E2F transcription factor, which is critical for the growth-deregulation and transforming properties of the viral E7 oncoprotein. In an attempt to apply interaction between the E7 oncoprotein and a target cellular protein Rb for an in vitro screening system for drugs against human papillomavirus infection, we primarily investigated the E7Rb binding through a pull down assay and enzyme-linked immunosorbent assay. The pull down assay showed that both glutathione S-transferase-tagged E7 and His-tagged E7 immobilized on resins specifically produced complexes with bacterially expressed Rb in a dose-dependent manner, as determined by immunoblot analyses. This result coincided with that of an enzyme-linked immunosorbent assay, which is a useful system for the mass screening of potential drugs. Taken together, this screening system (based on the interaction between E7 and Rb) can be a promising system in the development of drugs against cervical cancers caused by human papillomavirus infection.

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Development of Process-centered Performance Task For Curriculum·Instruction·Assessment Alignment based on 2015 Revised Home Economics Curriculum (교육과정, 수업, 평가의 일체화를 위한 가정과 과정 중심 수행과제 개발)

  • Lee, Gyeong Suk;Yoo, Taemyung;Yang, Ji Sun
    • Journal of Korean Home Economics Education Association
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    • v.30 no.4
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    • pp.165-185
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    • 2018
  • The purpose of the study was to show the development process of the total instructional alignment based on the 2015 revised technology home economics curriculum, and to present the detailed aspects of this task. The results of this study were as follows: First, the clarity phase of the performance established the annual evaluation plan to ensure the consistency of the achievement standard, the learning objectives and the assessment objectives through an analysis of the curriculum. Second, the development phase of the performance tasks and the grading criteria analyzed the contents area and determined the performance assessment task. The performance assessment consisted of a portfolio and project activity types, and also comprised of knowledge markets, as well as e-NIE tasks, including individual tasks and collective tasks. Third, the performance assessment and the results phase were conducted after planning the class operation according to the instructors' directions and arrangement of the schedule into block classes. Afterwards, the performance assessment tasks were revised by the teaching-learning community and class reflection on practical application. As these results demonstrate, developing performance tasks that are centered on the learner's development processes can contribute to teachers' expertise and improvement of instruction.

The Effects of Somatid on the Cytotoxicity of Cancer Cells and Human Papillomavirus Type 16 E6 and E7 Oncogenes (생기액(生肌液)의 세포독성 및 자궁경부암 바이러스 (HPV 16 type) 암 유발인자 E6와 E7의 작용에 미치는 효과)

  • Joung, Ok;Cho, Young-Sik;Cho, Cheong-Weon;Lee, Kyung-Ae;Shim, Jung-Hyun;Cho, Min-Chul;Lee, Hong-Soo;Yeom, Young-Il;Kim, Sang-Bom;Park, Sue-Nie;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.44 no.4
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    • pp.340-346
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    • 2000
  • Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV types 16 and 18) and cervical cancer has been well known. An extract of natural products, named as Somatid, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. This Somatid inhibited the proliferation of human cervical cancer cell lines (C-33A, SiHa, CaSki) and HaCaT keratinocytes in a dose response manner, In vitro binding assay and ELISA showed that Somatid inhibited the in vitro biding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, Somatid inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by Somatid. Our data suggested that Somatid inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus can be used as a putative anti-HPV agent for the treatment of cervical carcinomas caused by HPV.

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