• Title/Summary/Keyword: Over-expression

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Effect of DPBll Gene for the Transcriptional Induction by DNA Damage During Cell Cycle in Saccharomyces cerevisiae (출아효모의 세포주기동안 DNA 상해에 의한 발현 유도에 미치는 DPB11 유전자의 영향)

  • 선우양일;임선희;배호정;김중현;김은아;김승일;김수현;박정은;김재우
    • Korean Journal of Microbiology
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    • v.38 no.2
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    • pp.96-102
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    • 2002
  • The S-phase checkpoint mechanisms response to DNA damage or inhibition of DNA replication for maintenance of genetic stability in eukaryotic cells. These roles include cell cycle control arrest at S-phase and Iranscriptional induction of repair genes. To characterize the defects of dpbll mutant for both these responses, we examined the over-expression effect of DPBll gene, the sensitivity to HU, MMS, and the transcriptional pattern by DNA damage agent for RNRS mRNA. RNRS transcript is induced in response to a wide variety of agents that either damage D7A directly through chemical modification or induce stress by blocking DNA synthesis. As results, dpbll-1 cells are sensitive to DNA damage agents and the level of RNR3 mRNA is reduced approximately 40% than wild type cells. Moreover, we found the same results in dpb2-1 cells. Therefore, we propose that DPB2 and DPBll act as a sensor of replication that coordinates the transcriptional and cell cycle responses to replication blocks.

Developing an Independent Rendering Tool to Control Render Layer in 3D Animation Files (3D 애니메이션 파일의 렌더링 레이어를 조절하기 위한 독립적인 렌더링 도구 개발)

  • Kim, Ki-Hong;Choi, Chul-Young;Chae, Eel-Jin
    • The Journal of the Korea Contents Association
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    • v.8 no.9
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    • pp.84-93
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    • 2008
  • Since 2005, the production of feature 3D animation has been growing up worldwide and technology of hardware and software has been advanced fully enough to get over the limitation of visual expression, so that rendering weight of the whole work process is getting more important. A final file in the state right before rendering increases. the data size gets bigger. Therefore, if such final file is rendered as it is, the production schedule may not be met. For, some needless time shall be spent to call and save a high-capacity file in amendment work and to execute the rendering entirely again in addition to amendment. In this study, Maya API [1]was used to study the efficient information amendment method of extracting the rendering layers' files in order to develop a tool that executes rendering by applying such method. As the result, compliance to production schedule that is important in animation production can be done through the independent rendering tool based layer rendering that can maximize the efficiency of amendment work.

Evaluation of Reservoir Drought Response Capability Considering Precipitation of Non-irrigation Period using RCP Scenario (RCP 시나리오에 따른 비관개기 누적강수량을 고려한 둑높이기 저수지의 미래 가뭄대응능력 평가)

  • Bang, JeHong;Lee, Sang-Hyun;Choi, Jin-Yong;Lee, Sung-Hack
    • Journal of The Korean Society of Agricultural Engineers
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    • v.59 no.1
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    • pp.31-43
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    • 2017
  • Recent studies about irrigation water use have focused on agricultural reservoir operation in irrigation period. At the same time, it is significant to store water resource in reservoir during non-irrigation period in order to secure sufficient water in early growing season. In this study, Representative Concentration Pathways (RCP) 4.5, 8.5 scenarios with the Global Climate Model (GCM) of The Second Generation Earth System Model (CanESM2) were downscaled with bias correlation method. Cumulative precipitation during non-irrigation season, October to March, was analyzed. Interaction between cumulative precipitation and carry-over storage was analyzed with linear regression model for ten study reservoirs. Using the regression model, reservoir drought response ability was evaluated with expression of excess and deficiency. The results showed that future droughts will be more severe than past droughts. Especially in case of non-exceedance probability of 10%, drought in southern region seemed to be serious. Nine study reservoirs showed deficiency range from 10% to 55%, which turned out to be vulnerable for future drought. Only Jang-Chan reservoir was secure for early growing season in spite of drought with deficiency of 8% and -2%. The results of this study represents current agricultural reservoirs have vulnerability for the upcoming drought.

Cloning and Expression of a Thermostable ${\alpha}$-Galactosidase from the Thermophilic Fungus Talaromyces emersonii in the Methylotrophic Yeast Pichia pastoris

  • Simila, Janika;Gernig, Anita;Murray, Patrick;Fernandes, Sara;Tuohy, Maria G.
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1653-1663
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    • 2010
  • The first gene (${\alpha}$-gal1) encoding an extracellular ${\alpha}$-Dgalactosidase from the thermophilic fungus Talaromyces emersonii was cloned and characterized. The ${\alpha}$-gal1 gene consisted of an open reading frame of 1,792 base pairs interrupted by six introns that encoded a mature protein of 452 amino acids, including a 24 amino acid secretory signal sequence. The translated protein had highest identity with other fungal ${\alpha}$-galactosidases belonging to glycosyl hydrolase family 27. The ${\alpha}$-gal1 gene was overexpressed as a secretory protein with an N-terminal histidine tag in the methylotrophic yeast Pichia pastoris. Recombinant ${\alpha}$-Gal1 was secreted into the culture medium as a monomeric glycoprotein with a maximal yield of 10.75 mg/l and purified to homogeneity using Hisbinding nickel-agarose affinity chromatography. The purified enzyme was maximally active at $70^{\circ}C$, pH 4.5, and lost no activity over 10 days at $50^{\circ}C$. ${\alpha}$-Gal1 followed Michaelis-Menten kinetics ($V_{max}\;of\;240.3{\mu}M/min/mg,\;K_m\;of\;0.294 mM$) and was inhibited competitively by galactose ($K_m{^{obs}}$ of 0.57 mM, $K_i$ of 2.77 mM). The recombinant T. emersonii ${\alpha}$-galactosidase displayed broad substrate preference, being active on both oligo- and polymeric substrates, yet had strict specificity for the ${\alpha}$-galactosidic linkage. Owing to its substrate preference and noteworthy stability, ${\alpha}$-Gal1 is of particular interest for possible biotechnological applications involving the processing of plant materials.

XML Document Retrieval Models for Heterogeneous Data Set using Independent Regular paths (독립적인 질의 경로들을 사용하여 이질적인 문서들을 검색하는 XML 문서 검색 모델)

  • 유신재;민경섭;김형주
    • Journal of KIISE:Software and Applications
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    • v.30 no.1_2
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    • pp.140-152
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    • 2003
  • An XML document has a structure which may be irregular. It is difficult for end-users to comprehend the irregular document structure exactly. For these XML documents, an end-user has a difficulty in using structured query. Therefore, an end-user formulates no structured query or a query which has a little structure information. In this context, we propose new retrieval models which use the structured information for ranking and compensate the difference between user query structure and document structure. To ease with querying, we assume the independence among querying paths which represent structural constraints. Since this assumption makes degradation of the expression power of a query language, we also propose a model which overcome this problem. As there had been no test collections for XML documents, we made a small test collection from TIPSTER of the RTEC and experimented on this collection without a structured query, From this experiment, we showed that our models improve average precision about 67% over conventional Vector-Space model.

Defense Genes Induced by Pathogens and Abiotic Stresses in Panax ginseng C.A. Meyer

  • Lee, Ok-Ran;Sathiyaraj, Gayathri;Kim, Yu-Jin;In, Jun-Gyo;Kwon, Woo-Seang;Kim, Ju-Han;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.35 no.1
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    • pp.1-11
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    • 2011
  • Korean ginseng is a medicinally important perennial herb from the family Araliaceae. It has been cultivated for its highly valued medicinal properties for over 1,000 years in east Asian countries such as China, Korea, and Japan. Due to its longtime cultivation in shady areas, ginseng is frequently exposed to pathogenic infections. Plants protect themselves from microbial pathogens using an array of defense mechanisms, some of which are constitutively active, while others are activated upon pathogen invasion. These induced defense responses, controlled by defense-related genes, require tradeoffs in terms of plant fitness. We hypothesize that ginseng, as with other plants, possesses regulatory mechanisms that coordinate the activation of attacker-specific defenses in order to minimize fitness costs while attaining optimal resistance. Several classes of defense-related genes are induced by infection, wounds, irradiation, and other abiotic stresses. Both salicylates and jasmonates have been shown to cause such responses, although their specific roles and interactions in signaling and development are not fully understood in ginseng. This review summarizes possible defense-related genes in ginseng based on their expression patterns against biotic and abiotic stresses and describes their functional roles.

Cloning, Expression, and Characterization of UDP-glucose Pyrophosphorylase from Sphingomonas chungbukensis DJ77

  • Yoon, Moon-Young;Lee, Kyoung-Jin;Park, Hea-Chul;Park, Sung-Ha;Kim, Sang-Gon;Kim, Sung-Kun;Choi, Jung-Do
    • Bulletin of the Korean Chemical Society
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    • v.30 no.6
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    • pp.1360-1364
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    • 2009
  • The bacterium Sphingomonas chungbukensis DJ77 produces the extracellular polysaccharide gellan in high yield. Gellan produced by this bacterium is widely used as a gelling agent, and the enzyme UDP-glucose pyrophosphorylase (UGP) is thought to play a key role in the gellan biosynthetic pathway. The UGP gene has been successfully cloned and over-expressed in E. coli. The expressed enzyme was purified with a molecular weight of approximately 32 kDa, as determined by a SDS-polyacrylamide gel, but the enzyme appears as ca. 63 kDa on a native gel, suggesting that the enzyme is present in a homodimer. Kinetic analysis of UDP-glucose for UGP indicates $K_m$ = 1.14 mM and $V_{max}$ = 10.09 mM/min/mg at pH 8.0, which was determined to be the optimal pH for UGP catalytic activity. Amino acid sequence alignment against other bacteria suggests that the UGP contains two conserved domains: An activator binding site and a glucose-1-phosphate binding site. Site-directed mutagenesis of Lys194, located within the glucose-1-phosphate binding site, indicates that substitution of the charge-reversible residue Asp for Lys194 dramatically impairs the UGP activity, supporting the hypothesis that Lys194 plays a critical role in the catalysis.

Inducing apoptosis by the inhibition of c-myb in oral squamous carcinoma cell line, KB cell

  • Lee, Jung-Chang;Moon, Hyun-Ju;Lee, Young-Hee;Jung, Ji-Eun;Sharma, Manju;Jhee, Eun-Jung;Yi, Ho-Keun
    • International Journal of Oral Biology
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    • v.32 no.4
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    • pp.127-133
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    • 2007
  • Oral squamous cell carcinoma (OSCC) is the most common malignancy and is a major cause of worldwide cancer mortality. The proto-oncogene c-myb plays an important role in regulation of cell growth and differentiation, and it is expressed at high levels in hematopoietic cells and many other types of cancers. However, the function of c-myb is not well known in OSCC. The present study aimed to reveal the function of c-myb and to test the alternation of cell growth and signaling by c-myb in OSCC. In this study, c-myb and dominant-negatibe myb(DNmyb) were expressed in an adenovirus-mediated gene delivery system to KB cells. The over-expressed c-myb brought increased cellular proliferation compared with control cells. However, DN-myb infected KB cells showed significant reduction of cell growth and enhanced induction of apoptosis to activate PARP and caspase 9. c-myb induced increase of IGF-I, -II and IGF-IR expressions while DN-myb down-regulated these expression. Activation of ERK and Akt/PKB pathway was shown only in c-myb transduced cells. These findings suggest that the role of c-myb in cell growth of oral cancer cells is partially mediated through the modulation of IGFs, ERK and Akt/PKB. From this results, DN-myb is strongly recommended as a curable gene for the treatment of c-myb dependent malignancies such as OSCC.

Brazilian Test of Concrete Specimens Subjected to Different Loading Geometries: Review and New Insights

  • Garcia, Victor J.;Marquez, Carmen O.;Zuniga-Suarez, Alonso R.;Zuniga-Torres, Berenice C.;Villalta-Granda, Luis J.
    • International Journal of Concrete Structures and Materials
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    • v.11 no.2
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    • pp.343-363
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    • 2017
  • The objective of this work was finding out the most advisable testing conditions for an effective and robust characterization of the tensile strength (TS) of concrete disks. The independent variables were the loading geometry, the angle subtended by the contact area, disk diameter and thickness, maximum aggregate size, and the sample compression strength (CS). The effect of the independent variables was studied in a three groups of experiments using a factorial design with two levels and four factors. The likeliest location where failure beginning was calculated using the equations that account for the stress-strain field developed within the disk. The theoretical outcome shows that for failure beginning at the geometric center of the sample, it is necessary for the contact angle in the loading setup to be larger than or equal to a threshold value. Nevertheless, the measured indirect tensile strength must be adjusted to get a close estimate of the uniaxial TS of the material. The correction depends on the loading geometry, and we got their mathematical expression and cross-validated them with the reported in the literature. The experimental results show that a loading geometry with a curved contact area, uniform load distribution over the contact area, loads projected parallel to one another within the disk, and a contact angle bigger of $12^{\circ}$ is the most advisable and robust setup for implementation of BT on concrete disks. This work provides a description of the BT carries on concrete disks and put forward a characterization technique to study costly samples of cement based material that have been enabled to display new and improved properties with nanomaterials.

High Level of Soluble Expression in Escherichia coli and Characterisation of the Cloned Bacillus thuringiensis Cry4Ba Domain III Fragment

  • Chayaratanasin, Poramed;Moonsom, Seangdeun;Sakdee, Somsri;Chaisri, Urai;Katzenmeier, Gerd;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.40 no.1
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    • pp.58-64
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    • 2007
  • Similar to the other known structures of Bacillus thuringiensis Cry $\delta$-endotoxins, the crystal structure of the 65-kDa activated Cry4Ba toxin comprises three domains which are, from the N- to C-terminus, a bundle of $\alpha$-helices, a three-$\beta$-sheet domain, and a $\beta$-sandwich. To investigate the properties of the C-terminal domain III in isolation from the rest of the toxin, the cloned Cry4Ba-domain III was over-expressed as a 21-kDa soluble protein in Escherichia coli, which cross-reacted with anti-Cry4Ba domain III monoclonal antibody. A highly-purified domain III was obtained in a monomeric form by ion-exchange and size-exclusion FPLC. Circular dichroism spectroscopy indicated that the isolated domain III fragment distinctly exists as a $\beta$-sheet structure, corresponding to the domain III structure embodied in the Cry4Ba crystal structure. In vitro binding analysis via immuno-histochemical assay revealed that the Cry4Ba-domain III protein was able to bind to the apical microvilli of the susceptible Stegomyia aegypti larval midguts, albeit at lower-binding activity when compared with the full-length active toxin. These results demonstrate for the first time that the C-terminal domain III of the Cry4Ba mosquito-larvicidal protein, which can be isolated as a native folded monomer, conceivably participates in toxin-receptor recognition.