• Title/Summary/Keyword: Fusion Activity

Search Result 580, Processing Time 0.027 seconds

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
    • /
    • v.15 no.6 s.73
    • /
    • pp.961-967
    • /
    • 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.

Physiological Difference of Estrus and Pregnant Ovary in Korean Water Deer (한국산 고라니의 발정기와 임신기 난소의 생리학적 차이)

  • Kim, Sang-Hwan;Lee, Ho-Jun;Lee, Joo-Yong;Park, Yong-Su;Yoon, Jong-Taek
    • Journal of Embryo Transfer
    • /
    • v.29 no.1
    • /
    • pp.43-50
    • /
    • 2014
  • To describe the macroscopic anatomy and ovarian-physiological difference of the genital organs of the female Korean water deer, organs from captured animals in a wild area of Korea were dissected. The ovary of estrus group was about $1.10{\pm}0.02mm$ long and weighed about $0.50{\pm}0.02g$. And pregnant group was about $1.3{\pm}0.10mm$ long and weighed about $0.40{\pm}0.05g$. And the crowns of corpora lutea were found in the estrus group, but we couldn't find crowns at the pregnant group. Especially, the estrus ovaries tended (p=0.04) to be heavier than the ovaries during pregnancy. The MMP-9 activity was higher at the Graafian follicles of pregnant group than that in estrus group. However, with regard to follicles of estrus group, MMP-2 level was higher than that in pregnant group. Furthermore, apoptosis detection marker (Casp-3) was highly expressed in Graafian follicle of the pregnant group and the corpora lutea of estrus group. Thus, the differential expression of MMPs observed in this study suggests that the reflected the mechanisms underlying of monovulatory in estrus and/or pregnancy. Our results may be very useful as it provides with information that may be considered for the development of reproductive biotechnologies in endangered animals.

A Study on Revitalization of Dental Medical Tourism in Korea

  • Kim, Hyo-Kyung
    • Journal of the Korea Society of Computer and Information
    • /
    • v.25 no.10
    • /
    • pp.249-253
    • /
    • 2020
  • Medical tourism refers to visiting overseas to receive medical services and tourism at the same time. In other words, it can be said that it is a tourism activity that combines medical service, various cultural activities, leisure, and shopping. As such, medical tourism is not limited to simple medical services, but is a new high-value-added industry that creates new profits by fusion with tourism services. Many countries are making continuous efforts to foster their own tourism industry by attracting foreign tourists. Therefore, in order to secure competitiveness in dental services among the medical tourism industry in Korea, this study was to examine the relationship between dentist's treatment, hospital facilities, dental staff, and patient satisfaction. As a result, it was found that the dentist's treatment had a positive effect on both the staff of the dental facility in the hospital and the satisfaction of the patient.

Method of Service Curation based on User Log Analysis (사용자 이용로그 분석에 기반한 서비스 큐레이션 방법)

  • Hwang, Yun-Young;Kim, Dou Gyun;Kim, Bo-Ram;Park, Seong-Eun;Lee, Myunggyo;Yoon, Jungsun;Suh, Dongjun
    • Journal of Digital Contents Society
    • /
    • v.19 no.4
    • /
    • pp.701-709
    • /
    • 2018
  • Our research team implemented and operated the system by analyzing the membership information and identifying the different preferences for each group and providing the results of the recommendation based on accumulated membership information and activity log data to the individual. The utilization log was followed up. We analyzed how many people use recommended services and analyzed whether there are any factors other than the personalization service algorithm that affect the service utilization of the system with personalization. In addition, we propose recommendation methods based on behavioral changes when incentives are given through analyzing patterns of users' usage according to methods of recommending services and contents that are often used based on analysis contents.

A study on the development of muti-media system for road guideline (길안내를 위한 멀티미디어 개발에 관한 연구)

  • Jung Ho-Young;Lee In-Won
    • The Journal of The Korea Institute of Intelligent Transport Systems
    • /
    • v.3 no.1 s.4
    • /
    • pp.97-107
    • /
    • 2004
  • As one of the most serious urban problems, routine city traffic congestion causes not only unpleasantness but also damage with increases of road tolls. Recently, many TDM plans are being improved along with plans to more effectively utilize established facilities and maximize the effect. To achieve the purpose of Intelligent Transport Systems(ITS). Recently, the core of ITS is Advanced Traveler information Systems (ATIS.) that are activity researched. A development of various muti-media system is very important of road guideline. First of all a business processing for offering travel information and give a consideration to the United states of various muti-media systems.

  • PDF

The Development of Convergence Teaching-Learning Program for the Clothing Section of Home Economic Focused on Up-cycling (업사이클링 주제의 가정과 의생활 영역 융합교육 프로그램 개발)

  • Yang, Hyun-Sook;Lee, Youn-Hee;Uh, Mi-Kyung
    • Journal of the Korea Fashion and Costume Design Association
    • /
    • v.19 no.2
    • /
    • pp.49-61
    • /
    • 2017
  • The purpose of this study was to develop an educational program designed to allow students to experience 'up-cycling' first-hand during class and understand its significance by helping them improving their practical problem-solving abilities. Teachers of home economics, Korean, and social studies to third grade middle schooler were asked to analyze the curriculums of their subjects. The analysis results were then used to identify common elements among the units, reconstruct the curriculums, and develop an integrated lesson program to offer integrated activities. Based on these, a lesson program was developed to make an eco-bag and running shoes under the theme of 'up-cycling' in the unit of 'environmentally-friendly clothing and mending of clothes' in the home economics subject. The results were as follows: First, a topical fusion lesson program was developed to integrate three subjects together. The development process involved the integration of the topic of up-cycling, a program model for integrated lessons, and a teaching and learning process plan for topical integrated education. Secondly, a lesson program for home economics was developed that was applicable to actual home economics lessons based on the topical integrated lesson program. The lesson domains were divided into large, medium, and small to create a ten-lesson teaching and learning process plan needed for the lessons, teaching materials that could be put to actual uses in lessons, and activity and evaluation logs for learners.

  • PDF

Generation and Characterization of Cell-Permeable Greem Fluorescent Protein Mediated by the Basic Domain of Human Immunodeficiency Virus Type 1 Tat

  • Park, Jin-Seu;Kim, Kyeong-Ae;Ryu, Ji-Yoon;Choi, Eui-Yul;Lee, Kil-Soo;Choi, Soo-Young
    • Journal of Microbiology and Biotechnology
    • /
    • v.10 no.6
    • /
    • pp.797-804
    • /
    • 2000
  • The human immunodeficiency virus type 1 (HIV-1) Tat is one of the viral gene products essential for HIV replication. The exogenous Tat protein is transduced through the plasma membrane and then accumulated in a cell. The basic domain of the Tat protein, which is rich in arginine and lysine residues and called the protein transduction domain (PTD), has been identified to be responsible for this transduction activity. To better understand the nature of the transduction mediated by this highly basic domain of HIV-1 Tat, the Green Fluorescent Protein (GFP) was expressed and purified as a fusion protein with a peptide derived from the HIV-1 Tat basic domain in Escherichia coli. The transduction of Tat-GFP into mammalian cells was then determined by a Western blot analysis and fluorescence microscopy. The cells treated with Tat-GFP exhibited dose- and time-dependent increases in their intracellular level of the protein. the effective transduction of denatured Tat-GFP into both the nucleus and the cytoplasm of mammalian cells was also demonstrated, thereby indicating that the unfolding of the transduced protein is required for efficient transduction. Accordingly, the availability of recombinant Tat-GFP can facilitate the simple and specific identification of the protein transduction mediated by the HIV-1 Tat basic domain in living cells either by fluorescence microscopy or by a fluorescence-activated cell sorter analysis.

  • PDF

Expression and Promoter Analyses of Pepper CaCDPK4 (Capsicum annuum calcium dependent protein kinase 4) during Plant Defense Response to Incompatible Pathogen

  • Chung, Eun-Sook;Oh, Sang-Keun;Park, Jeong-Mee;Choi, Do-Il
    • The Plant Pathology Journal
    • /
    • v.23 no.2
    • /
    • pp.76-89
    • /
    • 2007
  • CaCDPK4, a full-length cDNA clone encoding Capsicum annuum calcium-dependent protein kinase 4, was isolated from chili pepper (Capsicum annuum L.). Deduced amino acid sequence of CaCDPK4 shares the highest homology with tobacco NpCDPK8 and chickpea CaCDPK2 with 79% identity. Genomic blot analyses revealed that CaCDPK4 is present as a single copy in pepper genome, but it belongs to a multigene family. CaCDPK4 was highly induced when pepper plants were inoculated with an incompatible bacterial pathogen. Induced levels of CaCDPK4 transcripts were also detected in pepper leaves by the treatment of ethephon, an ethylene-inducing agent, and high-salt stress condition. The bacterial-expressed GST-CaCDPK4 protein showed to retain the autophosphorylation activity in vitro. GUS expression driven by CaCDPK4 promoter was examined in transgenic Arabidopsis containing transcriptional fusion of CaCDPK4 promoter. GUS expression under CaCDPK4 promoter was strong in the root and veins of the seedlings. GW (-1965) and D3 (-1377) promoters conferred on GUS expression in response to inoculation of an incompatible bacterial pathogen, but D4-GUS (-913) and DS-GUS (-833) did not. Taken together, our results suggest that CaCDPK4 can be implicated on signal transduction pathway of defense response against an incompatible bacterial pathogen in pepper.

A Novel Oxidative Stress-inducible Peroxidase Promoter and Its Applications to Production of Pharmaceutical Proteins in Transgenic Cell Cultures

  • Lee, Ok-Sun;Park, Sun-Mi;Kwon, Suk-Yoon;Lee, Haeng-Soon;Kim, Kee-Yeun;Kim, Jae-Whune;Kwak, Sang-Soo
    • Journal of Plant Biotechnology
    • /
    • v.4 no.4
    • /
    • pp.143-150
    • /
    • 2002
  • A strong oxidative stress-inducible peroxidase promoter (referred to as SWPA2 promoter) was cloned from tell cultures of sweetpotato (Ipomoea batatas) and characterized in transgenic tobacco cultured cells in terms of biotechnological applications. Employing a transient expression assay in tobacco protoplasts, with five different 5'-deletion mutants of the SWPA2 promoter fused to the $\beta$-glucuronidase (GUS) reporter gene, the 1314 bp deletion mutant showed approximately 30 times higher GUS expression than the CaMV 35S promoter. The expression of GUS activity in suspension cultures of transgenic cells derived from transgenic tobacco leaves containing the -1314 bp SWPA2 promoter-GUS fusion was strongly expressed following 15 days of subculture compared to other deletion mutants, suggesting that the 1314 bp SWPA2 promoter will be biotechnologically useful for the development of transgenic cell lines engineered to produce key pharmaceutical proteins. In this respect, we developed transgenic cell lines such as tobacco (Nicotiana tabacum L. BY-2), ginseng (Panax ginseng) and Siberian ginseng (Acanthopanax senticosus) using a SWPA2 promoter to produce a human lactoferrin (hLf) and characterized the hLf production in cultured cells. The hLf production monitored by ELISA analysis in transgenic BY-2 cells was directly increased proportional to cell growth and reached a maximal level (up to 4.3% of total soluble protein) at the stationary phase in suspension cultures. The SWPA2 promoter should result in higher productivity and increased applications of plant cultured cells for the production of high-value recombinant proteins.

Misfolding-assisted Selection of Stable Protein Variants Using Phage Displays

  • Shin, Jong-Shik;Ryu, Seung-Hyun;Lee, Cheol-Ju;Yu, Myeong-Hee
    • BMB Reports
    • /
    • v.39 no.1
    • /
    • pp.55-60
    • /
    • 2006
  • We describe a phage display strategy, based on the differential resistance of proteins to denaturant-induced unfolding, that can be used to select protein variants with improved conformational stability. To test the efficiency of this strategy, wild-type and two stable variants of ${\alpha}_1$-antitrypsin (${\alpha}_1AT$) were fused to the gene III protein of M13 phage. These phages were incubated in unfolding solution containing denaturant (urea or guanidinium chloride), and then subjected to an unfavorable refolding procedure (dialysis at $37^{\circ}C$). Once the ${\alpha}_1AT$ moiety of the fusion protein had unfolded in the unfolding solution, in which the denaturant concentration was higher than the unfolding transition midpoint ($C_m$) of the ${\alpha}_1AT$ variant, around 20% of the phage retained binding affinity to anti-${\alpha}_1AT$ antibody due to a low refolding efficiency. Moreover, this affinity reduced to less than 5% when 10 mg/mL skimmed milk (a misfolding-promoting additive) was included during the unfolding/refolding procedure. In contrast, most binding affinity (>95%) remained if the ${\alpha}_1AT$ variant was stable enough to resist unfolding. Because this selection procedure does not affect the infectivity of M13, the method is expected to be generally applicable to the high-throughput screening of stable protein variants, when activity-based screening is not possible.