• Title/Summary/Keyword: fusion partner

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Development of human tumor necrosis factor-α muteins with improved therapeutic potential

  • Jang, Seung-Hwan;Kim, Hyo-Jin;Cho, Kwang-Hwi;Shin, Hang-Cheol
    • BMB Reports
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    • v.42 no.5
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    • pp.260-264
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    • 2009
  • Tumor necrosis factor-$\alpha$ (TNF-$\alpha$) exhibits cytotoxicity towards various tumor cells in vitro and induces apoptotic necrosis in transplanted tumors in vivo. It also shows severe toxicity when used systemically for the treatment of cancer patients, hampering the development of TNF-$\alpha$ as a potential anticancer drug. In order to understand the structure-function relation of TNF-$\alpha$ with respect to receptor binding, we selected four regions on the bottom of the TNF-$\alpha$ trimer that are in close contact with the receptor and carried out mutagenesis studies and computational modeling. From the study, various TNF-$\alpha$ muteins with a high therapeutic index were identified. These results will provide a structural basis for the design of highly potent TNF-$\alpha$ for therapeutic purposes. By conjugating TNF-$\alpha$ muteins with a high therapeutic index to a fusion partner, which targets a marker of angiogenesis, it could be possible to develop TNF-$\alpha$ based anticancer drugs.

Case Analysis on Platform Business Models for IT Service Planning (IT서비스 기획을 위한 플랫폼 비즈니스 모델 사례 분석연구)

  • Kim, Hyun Ji;Cha, yun so;Kim, Kyung Hoon
    • Korea Science and Art Forum
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    • v.25
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    • pp.103-118
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    • 2016
  • Due to the rapid development of ICT, corporate business models quickly changed and because of the radical growth of IT technology, sequential or gradual survival has become difficult. Internet-based new businesses such as IT service companies are seeking for new convergence business models that have not existed before to create business models that are more competitive, but the economic efficiency of business models that were successful in the past is wearing off. Yet, as reaching the critical point where the platform value becomes extremely high for platforms via the Internet is happening at a much higher speed than before, platform-ization has become a very important condition for rapid business expansion for all kinds of businesses. This study analyzes the necessity of establishing platform business models in IT service planning and identifies their characteristics through case analyses of platform business models. The results derived features First, there is a need to ensure sufficient buyers and sellers, and second, platform business model should provide customers with distinctive value of the only platforms are generating. third, the common interests between platform-driven company and a partner, participants Should be existing. Fourthly, by expanding base of participants and upgrades, expansion of adjacent areas we must have a continuous scalability and evolution must be sustainable. While it is expected that the identified characteristics will cause tremendous impacts to the establishment of platform business models and to the graphing of service planning, we also look forward to this study serving as the starting point for the establishment of theories of profit models for platform businesses, which were not mentioned in the study, so that planners responsible for platform-based IT service planning will spend less time and draw bigger schemes in building planning drafts.

Eine typologische Studio zur sprachlichen $Repr\"{a}sentation$ der komitativen und instrumentalen Relationen am Beispiel des Deutschen und Koreanischen (동반관계와 도구관계의 표현양식에 대한 언어유형학적 연구 - 독일어와 한국어를 중심으로 -)

  • Shin Yong-Min
    • Koreanishche Zeitschrift fur Deutsche Sprachwissenschaft
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    • v.10
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    • pp.201-229
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    • 2004
  • In der vorliegenden Albeit wird die funktionale $Dom\"{a}ne$ der Konkomitanz unter funktional-typologischen Gesichtpunkten untersucht. Dabei wird ein funktionales Rahmenwerk fur die Untersuchungen $\"{u}ber$ die verschiedenen komitativen und intrumentalen Relationen entwickelt. Es gibt eine Reihe von Partizipantenrelationen in der funktionalen $Dom\"{a}ne$ der Konkomitanz, $\n\"{a}mlich$ 'Partner', 'Companion', 'Vehicle', 'Tool', 'Material' und 'Manner'. Diese Partizipanten $k\"{o}nnen$ als Konkomitant bezichnet werden. Sie bilden ein Kontinuum $bez\"{u}glich$ der Empathiehierarchie sowie der Kontrollhierarchie. Konkomitanten $k\"{o}nnen$ bei ihrer syntaktischen Kodierung $gem\"{\a}{\ss}$ den spezifischen Typen der konkomitanten Relationen und ihren absoluten Eigenschaften variieren. Es werden sieben verschiedene Typen der Kodierungsstrategien vorgestellt, d.h. konkomitante $Pr\'{a}dikation$, adpositionale Markierung, Kasusmarkierung, Verbderivation, inkorporation, Konversion und lexikale Fusion. In einer gegebenen Sprache kann es $nat\"{u}rlich$ feinere Variation, die z.B. mit den Graden der Grammatikalisierung und Lexikakisierung dieser Kodierungsstrategien zu tun hat, geben. Die Distribution der strukturellen Apparaturen auf der funktionalen $Dom\"{a}ne$ der Konkomitanz ist innerhalb einer Sprache nicht einheitlich. Dies gilt auch $f\"{u}r$ den Sprachen (z.B. Deutsch und Englisch), die gewohnlich als ein typisches Beispiel fur den Synkretismus zwischen Komitativ und Instrumental angesehen werden. Die $Pr\"{a}position$ mit im Deutschen $dr\"{u}ckt$ nur eine Teilmenge der verschiedenen konkomitanten Relationen aus. Tlansportrnittei' und Material' werden also nicht durch die $Pr\"{a}position$ mit $ausgedr\"{u}ckt$. Beim Ausdruck der konkomitanten Relationen verwendet Deutsch konsistent die Strategie der adpositionalen Markierung. Andere $m\"{o}gliche$ Strategien $k\"{o}nnen$ als ein minimaler Sonderfall angesehen werden. Koreanisch $verf\"{u}gt$ hingegen auf zwei Ausdrucksstrategien, $n\"{a}mlich$ konkomitante $Pr\"{a}dikation$ und Kasusmarkieriing. Die Auswahl zwischen den beiden Strategien ist meistens von den Empathieeigenschaften der Konkomitanten $abh\"{a}ngig$. Auch innerhalb einer Ausdrucksstrategie spielt die Empathie der Konkomitanten eine wichtige Rolle bei der Auswahl der Strukturmittel. Einen Synkretismus weisen im Koreanischen 'Partner' & 'Companion' einerseits und 'Vehicle' & 'Tool' & 'Material' andererseits auf, Die Ersteren werden durch die auditive Kasusmarkierung und die Letzteren durch die instrumentale Kasusmarkierung ausgedruckt. Ein Synkretimus beim Ausdruck der verschiedenen konkomitanten Relationen ist in den Sprachen, die nur konkomitante $Pr\"{a}dikation$ aufweisen, nicht zu erwarten, denn ein konkomitantes $Pr\"{a}dikat$ gibt die Art der Beteiligung der Konkomitanten an der Situation lexikalisch mehr oder weniger explizit wieder

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Surface Display of Heme- and Diflavin-Containing Cytochrome P450 BM3 in Escherichia coli: A Whole-Cell Biocatalyst for Oxidation

  • Yim, Sung-Kun;Kim, Dong-Hyun;Jung, Heung-Chae;Pan, Jae-Gu;Kang, Hyung-Sik;Ahn, Tae-Ho;Yun, Chul-Ho
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.712-717
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    • 2010
  • Cytochrome P450 enzymes (P450s) are involved in the synthesis of a wide variety of valuable products and in the degradation of numerous toxic compounds. The P450 BM3 (CYP102A1) from Bacillus megaterium was the first P450 discovered to be fused to its redox partner, a mammalian-like diflavin reductase. Here, we report the development of a whole-cell biocatalyst using ice-nucleation protein (Inp) from Pseudomonas syringae to display a hemeand diflavin-containing oxidoreductase, P450 BM3 (a single, 119-kDa polypeptide with domains of both an oxygenase and a reductase) on the surface of Escherichia coli. The surface localization and functionality of the fusion protein containing P450 BM3 were verified by flow cytometry and measurement of enzymatic activities. The results of this study comprise the first report of microbial cell-surface display of a heme- and diflavin-containing enzyme. This system should allow us to select and develop oxidoreductases containing heme and/or flavins into practically useful whole-cell biocatalysts for extensive biotechnological applications, including selective synthesis of new chemicals and pharmaceuticals, bioconversion, bioremediation, live vaccine development, and biochip development.

Production of Recombinant Trehalose Synthase from Thermus caldophilus GK24 (재조합 내열성 트레할로스 합성효소의 생산)

  • Choi, Jae-Youl;Cha, Wol-Suk;Shin, Hyun-Jae
    • KSBB Journal
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    • v.21 no.4
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    • pp.298-301
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    • 2006
  • A gene(GeneBank AF 135796) coding for a trehalose synthase from Thermus caldophilus GK24 was cloned into Escherichia coli K12 using five vector systems. The constitutive expression system(pHCETS) which shows the highest trehalose synthase activity from flask culture of recombinant E. coli was selected for the production of trehalose from maltose. For the shake flask culture, the final dry cell weight was 0.9 g/L and the trehalose synthase activity was 25 U/mL. Fed-batch culture of recombinant E. coli harboring plasmid pHCETS which uses the glycerolas a carbon source was performed in jar fermentor: the dry cell weight of 20 g/L and the trehalose synthase activity of 13.7 U/mL were attained in 48 h.

A Study on the RPA Module Implementation of Cloud Travel and Expense Management System (클라우드 경비지출관리 솔루션의 RPA 모듈 구현에 관한 연구)

  • Lee, In-Sung;Oh, In-Ha
    • Journal of Convergence for Information Technology
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    • v.11 no.4
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    • pp.46-54
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    • 2021
  • As the realization of the 4th industrial revolution is approaching, the paradigm of the corporate work environment is changing to digital, from the traditional work environment. In particular, technologies like RPA(robotic process automation) and chatbot reduce the need for human labor or task by automating simple repetitive tasks, enabling humans to focus on more valuable tasks. In this study, corporates operating expense management services in public cloud computing environments develop a cloud module that simplifies expense report management by grafting robotic process automation and chatbot technology. According to the result of the expert evaluation, the developed system marked 80.3% of satisfaction levels and the highest satisfaction level 94% was confirmed in terms of easy of use. Based on the research result, future research can be suggested to expand the works occurring inside and outside the company to a single RPA environment by additionally linking the work system related to expense management.

Osteonectin Interacts with Human Nebulin C-terminus in Skeletal Muscle

  • Park, Eun-Ran;Kim, Hyun-Suk;Choi, Jun-Hyuk;Lee, Yeong-Mi;Choi, Jae-Kyoung;Joo, Young-Mi;Ahn, Seung-Ju;Min, Byung-In;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.263-272
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    • 2007
  • Nebulin is a giant actin binding protein (600-900 kDa) which is specific to skeletal muscle. This protein is known to regulate thin filaments length in sarcomere as a molecular template. The C-terminus of nebulin is located in the Z-disc of muscle sarcomere and is bound to other proteins such like myopalladin, titin, archvillin, and desmin. The N-terminus of nebulin binds to tropomodulin at the pointed ends of the thin filaments. In recent research, nebulin not only found in brain but also expressed in heart, stomach, and liver. So, the roles of nebulin in non-muscle tissue have been studied. However, lack of information or studies on nebulin binding proteins and nebulin function in brain are available so far. Therefore, the current study have investigated a novel binding partner of Nebulin C-terminus by using yeast two-hybrid screening with human brain cDNA library. Nebulin C-terminus, containing simple repeats, serine rich and SH3 domain, interacts with osteonectin C-terminal region. The specific interaction of nebulin and osteonectin were confirmed in vitro by using GST pull-down assay and reconfirmed in vivo by using transfected COS-7 cells with EGFP-tagged nebulin and DsRed-tagged osteonectin. Consequently, this study identified SH3 domain in nebulin C-terminus specifically binds to extracellular Ca-binding (EeC domain in osteonectin. Also, nebulin C-terminus fusion protein colocalized with osteonectin EC domain fusion protein in transfected COS-7 cells. The current study found the interaction between nebulin and osteonectin in human brain for the first time and suggested the nebulin in brain may be associated with osteonectin, as a regulator of cell cycle progression and mitosis.

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Expression and Purification of Recombinant Human Interferon-gamma Produced by Escherichia coli (대장균이 생산한 재조합 인체 감마인터페론의 발현과 정제)

  • Park, Jung-Ryeol;Kim, Sung-Woo;Kim, Jae-Bum;Jung, Woo-Hyuk;Han, Myung-Wan;Jo, Young-Bae;Jung, Joon-Ki
    • KSBB Journal
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    • v.21 no.3
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    • pp.204-211
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    • 2006
  • For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

A Research and Application of Polyhydroxyalkanoates in Biosensor Chip (생분해성 고분자, 폴리하이드록시알카노에이트를 이용한 바이오센서 칩 연구와 그 응용)

  • Park, T.J.;Lee, S.Y.
    • KSBB Journal
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    • v.22 no.6
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    • pp.371-377
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    • 2007
  • Polyhydroxyalkanoates (PHAs) are a family of microbial polyesters that can be produced by fermentation from renewable resources. PHAs can be used as completely biodegradable plastics or elastomers. In this paper, novel applications of PHAs in biosensor are described. A general platform technology was developed by using the substrate binding domain (SBD) of PHA depolymerase as a fusion partner to immobilize proteins of interest on PHA surface. It could be shown that the proteins fused to the SBD of PHA depolymerase could be specifically immobilized onto PHA film, PHA microbead, and microcontact printed PHA surface. We review the results obtained for monitoring the specific interaction between the SBO and PHA by using enhanced green fluorescent protein, red fluorescent protein, single chain antibody against hepatitis B virus preS2 surface protein and severe acute respiratory syndrome coronavirus surface antigen as model proteins. Thus, this system can be efficiently used for studying protein-protein and possibly protein-biomolecule interactions for various biotechnological applications.

Some characters of bacterial cellulases in goats' rumen elucidated by metagenomic DNA analysis and the role of fibronectin 3 module for endoglucanase function

  • Nguyen, Khanh Hoang Viet;Dao, Trong Khoa;Nguyen, Hong Duong;Nguyen, Khanh Hai;Nguyen, Thi Quy;Nguyen, Thuy Tien;Nguyen, Thi Mai Phuong;Truong, Nam Hai;Do, Thi Huyen
    • Animal Bioscience
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    • v.34 no.5
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    • pp.867-879
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    • 2021
  • Objective: Fibronectin 3 (FN3) and immunoglobulin like modules (Ig) are usually collocated beside modular cellulase catalytic domains. However, very few researches have investigated the role of these modules. In a previous study, we have sequenced and analyzed bacterial metagenomic DNA in Vietnamese goats' rumen and found that cellulase-producing bacteria and cellulase families were dominant. In this study, the properties of modular cellulases and the role of a FN3 in unique endoglucanase belonging to glycosyl hydorlase (GH) family 5 were determined. Methods: Based on Pfam analysis, the cellulases sequences containing FN3, Ig modules were extracted from 297 complete open reading frames (ORFs). The alkaline, thermostability, tertiary structure of deduced enzymes were predicted by AcalPred, TBI software, Phyre2 and Swiss models. Then, whole and truncated forms of a selected gene were expressed in Escherichia coli and purified by His-tag affinity column for assessment of FN3 ability to enhance enzyme activity, solubility and conformation. Results: From 297 complete ORFs coding for cellulases, 148 sequences containing FN3, Ig were identified. Mostly FN3 appeared in 90.9% beta-glucosidases belonging to glycosyl hydrolase family 3 (GH3) and situated downstream of catalytic domains. The Ig was found upstream of 100% endoglucanase GH9. Rarely FN3 was seen to be situated downstream of X domain and upstream of catalytic domain endoglucanase GH5. Whole enzyme (called XFN3GH5 based on modular structure) and truncate forms FN3, XFN3, FN3GH5, GH5 were cloned in pET22b (+) and pET22SUMO to be expressed in single and fusion forms with a small ubiquitin-related modifier partner (S). The FN3, SFN3 increased GH5 solubility in FN3GH5, SFN3GH5. The SFN3 partly served for GH5 conformation in SFN3GH5, increased modules interaction and enzyme-soluble substrate affinity to enhance SXFN3GH5, SFN3GH5 activities in mixtures. Both SFN3 and SXFN3 did not anchor enzyme on filter paper but exfoliate and separate cellulose chains on filter paper for enzyme hydrolysis. Conclusion: Based on these findings, the presence of FN3 module in certain cellulases was confirmed and it assisted for enzyme conformation and activity in both soluble and insoluble substrate.