• Title/Summary/Keyword: Expression Form

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Induction of Apoptosis by N-methyl-N'-nitro-N-nitrosoguanidine, an Alkylating Agent, in Human Prostate Carcinoma Cells (인체 전립선 암세포에서 Alkylating Agent인 N-methyl-N'-nitro- N-nitrosoguanidine에 의한 Apoptosis유발)

  • Park, Cheol;Choi, Byung-Tae;Lee, Won-Ho;Choi, Yung-Hyun
    • Toxicological Research
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    • v.19 no.2
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    • pp.91-98
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    • 2003
  • Alkylating agents form alkylated base adducts in the DNA and cause DNA lesions leading to cell killing. In this study, we investigated the mechanism of apoptosis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) in PC-3 and DU145 human prostate carcinoma cell lines. MNNG treatment resulted in the inhibition of cell proliferation in a concentration-dependent manner to a similar extent in both cell lines. This anti-proliferative effect of PC-3 and DU145 cells by MNNG was associated with morphological changed such as membrane shrinking, cell rounding up and formation of apoptotic bodies. MNNG treatment also induced a proteolytic cleavage of specific target proteins such as poly(ADP-ribose) polymerase (PARP) and $\beta$-catenin proteins in DU145 cells but in PC-3 cells. Furthermore, we observed an increase of proapoptotic protein Bax family expression and a decrease of antiapoptotic protein Bcl-2 family by MNNG treatment in a concentration-dependent manner MNNG also induced a proteolytic activation of caspase-3 and -9, which is believed to play a central role in the apoptotic signaling pathway.

Construction of ELISA System for the Detection of Indian citrus ringspot virus (Indian citrus ringspot virus의 ELISA 진단 시스템 구축)

  • Shin, Myeung-Ju;Kwon, Young-Chul;Ro, Hyeon-Su;Lee, Hyun-Sook
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.231-235
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    • 2012
  • Indian citrus ring spot virus (ICRSV) is known to cause a serious disease to citrus, especially to Kinnow mandarin, the popular cultivated citrus species in India. In this study, we developed diagnostic systems based on enzyme-linked immunosorbent assay (ELISA). In order to generate antibodies against ICRSV coat protein, we overexpressed the coat protein in Escherichia coli using the pET15b expression vector containing an optimized ICRSV coat protein gene. The recombinant ICRSV coat protein was overexpressed as soluble form at $37^{\circ}C$ upon IPTG induction. The protein was purified to 95% in purity by Ni-NTA column chromatography. The purified protein was immunized to rabbit for the generation of polyclonal antibody (PAb). The PAb showed a specific immunoreaction to recombinant ICRSV coat protein in western blot analysis and ELISA. Diluted rabbit antisera (10,000 fold) could detect less than 10 ng and 5 ng of the target protein in western blot and ELISA analysis, respectively.

Comparison of Accuracy of Stability Formulas for Breakwater Armor Stones (방파제 피복석 안정 공식의 정밀도 비교)

  • 서경덕;유동훈
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.15 no.3
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    • pp.186-189
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    • 2003
  • Various empirical formulas have been developed to describe the stability of breakwater armor stones. The Hudson formula is known to have needs to be refined in many ways. but it is most widely used because of its simplicity. The van der Meer formula is sometimes used for the cross-check, but it is not very popularly used due to its complexity and the uncertainty of the relevant parameters. Recently, on the other hand, Yoo(2003) proposed a formula of a simple form using a new non-dimensional number which is called 'action slope' (abbreviated as Yoo formula hereinafter). In this study, in order to provide coastal engineers with some idea of the use of these formulas, their accuracies were estimated by comparing with the experimental data reported by van der Meer in 1987 and 1988. It was found that the van der Meer formula showed the highest accuracy, while the Hudson formula of very low accuracy needed considered judgment on its use. On the other hand, the Yoo formula of slightly lower accuracy but simpler expression than the van der Meer formula seems to be able to be used in engineering practice if a slightly higher safety factor is taken.

Secretion and Localization of Pseudomonas auratiaca Levansucrase Expressed in Saccharomyces cerevisiae (Saccharomyces cerevisiae에서 발현된 Pseudomonas aurantiaca Levansucrase의 분비국재성)

  • 임채권;김광현;김철호;이상기;남수완
    • Microbiology and Biotechnology Letters
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    • v.32 no.3
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    • pp.206-211
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    • 2004
  • Levansucrase gene(lscA) from Pseudomonas aurantiaca was subcloned downstream of GAL1 promoter in pYES 2.0 and pYInu-AT [GAL10 promoter + exoinulinase signal sequence of Kluyveromyces marxianus], resulting pYES-lscA and p YInu-lscA, respectively. The two expression plasmids were introduced into an invertase-deficient strain, Sacchromayces cerevisiae SEY2102, and transformants with high activity of levansucrase were selected. When each yeast transform ants was cultivated in medium containing galactose, the extracellular and intracellular activities of levansucrase reached about 8.62 U/ml with the strain harboring pYES-lscA and 5.43 U/ml with the strain harboring pYInu-lscA. The levansucrase activity of 80% was detected in the periplasmic space and cytoplasm. The levansucrase activity in the medium of SEY2102/pYInu-lscA was 0.87 U/ml whereas that of SEY2102/pYES-lscA was 0.47 U/ml, which implying the exoinulinase signal sequence didn't enhance the secretion efficiency of levansucrase. Furthermore, the recombinant levansucrase expressed in yeast seems to be produced as a hyper-glycosylated form.

Simulacra Theory as a Conceptual Framework for Understanding Expression and Technology in (<수면의 과학>에 나타난 시뮬라시옹 표현기법 연구)

  • Bang, Yoon-Kyeong
    • Cartoon and Animation Studies
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    • s.24
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    • pp.135-154
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    • 2011
  • Simulacra theory as propounded by Jean Baudrillard in his seminal treatise, "Simulacra and Simulation," explores the significance of images, media and art in contemporary culture. Its central theoretical premise is the simulacrum, a sign or symbol that plays a crucial role in constructing perceived reality but which lacks a real-world referent. In Michel Gondry creates simulacra in the form of hallucinatory dream imagery by combining stop-motion animation and live-action elements. As experimental film-making that combines analog and minimum digital technologies, the result is a tour-de-force of synchronization. This study analyzes the film's technique and expressive content and, by adopting simulacra theory as a conceptual framework, aims to provide a better understanding of Gondry's work.

Production of Soluble Human Granulocyte Colony Stimulating Factor in E. coli by Molecular Chaperones

  • PARK SO-LIM;SHIN EUN-JUNG;HONG SEUNG-PYO;JEON SUNG-JONG;NAM SOO-WAN
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1267-1272
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    • 2005
  • The effects of coexpression of GroEL/ES and DnaK/DnaJ/GrpE chaperones on the productivity of the soluble form of human granulocyte colony stimulating factor (hG-CSF) in E. coli were examined. Recombinant hG-CSF protein was coexpressed with DnaK/DnaJ/GrpE or GroEL/ES chaperones under the control of the araB or Pzt-1 promoter, respectively. The optimal concentration of L-arabinose for the expression of DnaK/DnaJ/GrpE was found to be 1 mg/ml. When L-arabinose was added at $OD_{600}$=0.2 (early-exponential phase), soluble hG-CSF production was greatly increased. In addition, it was observed that the DnaK/DnaJ/GrpE and GroEL/ES chaperones had no synergistic effects on preventing aggregation of hG-CSF protein. Consequently, by coexpression of the DnaK/DnaJ/GrpE chaperone, the signal intensity of the hG-CSF protein band in the soluble fraction of cell lysate was increased from $3.5\%\;to\;13.9\%$, and Western blot analysis also revealed about a 4-5-fold increase of production of soluble hG-CSF over the non-induction case of DnaK/DnaJ/GrpE.

The Function of Social Criticism Film Implemented in Animation Film (애니메이션 영화 <창, 2012>에 구현된 시사고발 영화 기능)

  • Lee, Hyun-Seok
    • The Journal of the Korea Contents Association
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    • v.17 no.3
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    • pp.150-161
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    • 2017
  • The short animation directed by Sang-Ho Yeon has led the sympathy and reflection of audiences through reveling the violence of the military in the form of the social criticism film. The explores the social value of animation by the content of social criticism, not for entertainment uses but for public interest. This research addresses how the function of social criticism implements in animation film, especially, its narratives and direction. In the light of this, firstly, literature will review the general characteristics of social criticism films, Secondly, based on literature reviews, analysis tools will be drew, thirdly, the case study regarding will be processed by focusing on The function of social criticism, the technique of direction and expression of animation. As a result of this research, shows unique traits because of the hierarchical and exclusive characteristics of military culture, which differs from general characteristics of social criticism films dealing with historic and social issues. In addition to this, the function of social criticism is effectively implemented in animation film, by the creative narratives and technique of direction by a director, Sang-Ho Yeon.

Characterization of Surface Layer Proteins in Lactobacillus crispatus Isolate ZJ001

  • Chen, Xueyan;Chen, Yang;Li, Xiaoliang;Chen, Ning;Fang, Weihuan
    • Journal of Microbiology and Biotechnology
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    • v.19 no.10
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    • pp.1176-1183
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    • 2009
  • Lactobacillus crispatus (L. crispatus) ZJ001 is highly adhesive to epithelial cells and expresses S-layer proteins. In this study, S-S-layer layer genes were sequenced and expressed in E. coli to characterize the function of proteins with this particular strain. L. crispatus ZJ001 harbored two S-layer genes slpA and slpB, and only slpA gene was expressed in the bacterium, as revealed by RT-PCR and immunoassays. The mature SlpA showed 47% amino acid sequence identity to SlpB. The SlpA and SlpB of L. crispatus ZJ001 were highly homologous at the C-terminal region to other Lactobacillus S-layer proteins, but were substantially variable at N-terminal and middle regions. Electron microscopic analysis indicated that His-slpA expressed in E. coli was able to form a sheet-like structure similar to the natural S-layer, but His-slpB formed as disc-like structures. In the cell binding experiments, HeLa cells were able to bind to both recombinant His-slpA and His-slpB proteins to the extent similar to the natural S-layer. The cell binding domains remain mostly in the N-terminal regions in SlpA and SlpB, as shown by high binding of truncated peptides SlpA2-228 and SlpB2-249. Our results indicated that SlpA was active and high binding to HeLa cells, and that the slpA gene could be targeted to display foreign proteins on the bacterial surface of ZJ001 as a potential mucosal vaccine vector.

Molecular and Cellular Analyses of NCP, a Nuclear and Centrosomal Protein in Mouse Gametes and Early Embryos

  • Oh, Hwa-Soon;Youn, Hong-Hee;Lee, Kwang-Hee;Son, Chae-Ick;Lee, Sang-Ho
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.97-97
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    • 2003
  • For many animals the centrosome consists of a pair of centrioles and surrounding pericentriolar materials (PCMs). PCMs have been known to play roles during cell division. It is known that centrioles are necessary to assemble centrosomal components. However, many types of oocytes undergo meiosis without centrioles. It is known that in nonmurine mammalian species, the sperm introduces an intact proximal centriole unlike sea urchin where two centrioles are introduced. In case of mouse sperm, the presence of centrosome is not clear In this study, a monoclonal antibody was developed to investigate centrosome during mouse germ cell and early embryo development. Results of immunostaining and Western blotting in CHO cells suggest that the monoclonal antibody recognizes a nuclear and centrosomal protein, thus called NCP. The NCP monoclonal antibody was used to screen a cDNA expression library prepared from 12.5 mouse brain to isolate NCP gene. Nucleotide size of NCP gene obtained from immunoscreening was about 5.5kb. It is determined that the NCP may be closely related with pericentriolar material -1 gene (Pcm-1) from the result of sequencing analysis. The molecular weight, 66kDa, calculated by known DNA sequence in database is consistent with that of detected from Western blotting using CHO cell lysates. Therefore, it is assumed that NCP may be alternative splicing form of Pcm-1 of which molecular weight is 228kDa. In mouse oocytes, NCP was distributed in nucleus as in CHO cells. It was shown that the NCP was localized around neck region, probably the centrosome in mouse neck region. Interestingly, dramatic change in distribution of NCP was also shown in male germ cell development. Finally, we observed the cellular distribution of NCP during early embryo development. NCP was detected in nucleus as well as centrosome foci. It is suggested that the centrioles reassembly we occurring in blastocysts and then affects the distribution of NCP.

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A Study on the Form of Charms with the Attributes of Experimental Typography (실험 타이포그래피 관점에서의 부적의 조형성)

  • 정성환;김민호
    • Archives of design research
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    • v.14 no.4
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    • pp.99-108
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    • 2001
  • The Charm is composed of various kind of ancient signs, letters and systematic characters so that they delivers messages with many expressions. They, compared with the experimental typography, are more logical and experimental than we generally think. Also, they have meanings and attributes as characters do, in which there are symbols, hieroglyphs, pictures and other similar words. Each attribute diverse from symbolic meaning, associates to deconstruction of letters. It is needed to review the experimental attributes of charms with the view of structural typography for potentialities of expression. The purpose of the study is to analyze and classify the morphologic construction of charms so that we can find the possibilities of applying the attributes to visual languages and typography. Future study seems to be extending the area of expressing of typography fit for our own culture, not for Western

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