• Title/Summary/Keyword: Alternative expression

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Novel Vectors for the Convenient Cloning and Expression of In Vivo Biotinylated Proteins in Escherichia coli

  • Cho, Eun-Wie;Park, Jung-Hyun;Na, Shin-Young;Kim, Kil-Lyong
    • BMB Reports
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    • v.32 no.5
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    • pp.497-501
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    • 1999
  • Biotinylation of recombinant proteins is a powerful tool for the detection and analysis of proteins of interest in a large variety of assay systems. The recent development of in vivo biotinylation techniques in E. coli has opened new possibilities for the production of site-specifically biotinylated proteins without the need for further manipulation after the isolation of the recombinantly expressed proteins. In the present study, a novel vector set was generated which allows the convenient cloning and expression of proteins of interest fused with an N-terminal in vivo biotinylated thioredoxin (TRX) protein. These vectors were derived from the previously reported pBIOTRX vector into which was incorporated part of the pBluescript II+phagemid multiple cloning site (MCS), amplified by PCR using a pair of sophisticated oligonucleotide primers. The functionality of these novel vectors was examined in this system by recombinant expression of rat transforming growth factor-$\beta$. Western-blot analysis using TRX-specific antibodies or peroxidase-conjugated streptavidin confirmed the successful induction of the fusion protein and the in vivo conjugation of biotin molecules, respectively. The convenience of molecular subcloning provided by the MCS and the effective in vivo biotinylation of proteins of interest makes this novel vector set an interesting alternative for the production of biotinylated proteins.

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Expression Analysis of ${\beta}$-Ketothiolase and Acetoacetyl-CoA Reductase of Rhodobacter sphaeroides

  • KHO, DHONG HYO;CHEOL YUN JEONG;JEONG JUG LEE
    • Journal of Microbiology and Biotechnology
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    • v.11 no.6
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    • pp.1031-1037
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    • 2001
  • By a sequential action of ${\beta}$-ketothiolase and acetoacetyl-CoA reductase, two molecules of acetyl-CoA re converted into D-3-hydroxybutyryl-CoA, a substrate for PHB synthase to form poly-3-hydroxybutyryl-CoA, a substrate for PHB synthase to form poly-3-hydroxybutyrate (PHB) of rhodobacter sphaeroides. The ${\beta}$-ketothiolase gene, phbA, and acetoacetyl-CoA reductase gene, phbB, were cloned and analyzed for their expression. Enzyme activities of ${\beta}$-ketothiolase and acetoacetyl-CoA reductase showed constitutive levels during aerobic and photoheterotrophic growth of R. sphaeroides. In addition, no difference of each enzyme activity was observed between cells grown aerobically and photoheterotrophically. The constitutive level of the enzyme activities are regulated according to the growth phases along with growth conditions. Thus, phbAB expression is not determinative in regulating the PB content. On the other hand, phbA-deleted cell AZI accumulated only $10\%$ PHB of the wild-type, and an elevated dosage of phbAB in trans in R. sphaeroides resulted in a higher content of PHB, indicating that phbAB codes for the enzymes responsible for providing the main supply of subsyrate for PHB synthase. PHB formation by an alternative pathway that does not does not depend on the phbA-and phbB-coding enzymes is also proposed.

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The Study of Skeleton System for Facial Expression Animation (Skeleton System으로 운용되는 얼굴표정 애니메이션에 관한 연구)

  • Oh, Seong-Suk
    • Journal of Korea Game Society
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    • v.8 no.2
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    • pp.47-55
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    • 2008
  • This paper introduces that SSFE(Skeleton System for Facial Expression) to deform facial expressions by rigging of skeletons does same functions with 14 facial muscles based on anatomy. A three dimensional animation tool (MAYA 8.5) is utilized for making the SSFE that presents deformation of mesh models implementing facial expressions around eyes, nose and mouse. The SSFE has a good reusability within diverse human mesh models. The reusability of SSFE can be understood as OSMU(One Source Multi Use) of three dimensional animation production method. It can be a good alternative technique for reducing production budget of animations. It can also be used for three dimensional animation industries such as virtual reality and game.

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Soft decision for Gray Coded PAM Signals Using Max-Log-MAP (Max-Log-MAP을 이용한 Gray 부호화된 PAM 신호의 연판정 계산식)

  • Hyun, Kwang-Min;Yoon, Dong-Weon
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.31 no.2C
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    • pp.117-122
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    • 2006
  • In this paper, we present a simple and general soft bit decision expression for a Gray coded PAM signal over additive white Gaussian noise(AWGN) channel with the log likelihood ratio(LLR). In order to reduce the complexity of the LLR calculation, we make the bitwise LLR expression simple by replacing the mathematical max functions of the conventional Max-Log-MAP expression with simple arithmetic functions associated with some deterministic parameters, such as a received value and distance between symbols on a signal space. Taking the implementation issues, like the area of silicon, the power consumption, the timing latency, and so on, into consideration, we submit that the proposed method is a promising alternative way to conventional methods for reconfigurable systems.

The Study Characteristics of Korean Traditional Space from an Ecological and Aesthetic (생태미학적 관점에서 본 한국 전통공간의 특성에 관한 연구)

  • Yang, Eun-Jee;Kim, Kai-Chun
    • Korean Institute of Interior Design Journal
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    • v.19 no.3
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    • pp.57-66
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    • 2010
  • Currently, the nature and ecosystem's destruction by human beings and science is appearing as a serious problem. The possibility of ecological aesthetics is being sought as one of operations for new society in the contemporary society that the nature and ecosystem are getting attention, The present research aims to examine the characteristics and its expression method by setting the ecological aesthetics as an alternative of the future design. The history, meaning, mental image and image of Korean traditional Space forming the balance and harmony with the Nature, etc., which are important design concepts, were deduced as the beauty of emptiness, the beauty of a gap, the beauty of harmony and the beauty of changing and the beauty of simplicity as characteristics of experiencing space. If the deduced expression is examined through a case analysis of traditional housing, the characteristics trying to unite with the Nature while seeing ecology and aesthetics as one root, which the ecological aesthetics regarding as a root. The ecological arts based on the ecological aesthetics enters inside the universal life of human beings by examining the ecological and aesthetic characteristics appearing in Korean traditional Space, and the arts will move forward in a direction that its function and role link with maintenance of total ecosystem. The space application possibility and expression characteristics of ecological aesthetics should be continuously researched.

Repetitive Somatic Embryogenesis in Cacao and Optimisation of Gene Expression by Particle Bombardment

  • Marcelo de Oliveira, Santos;Albuquerque de Barros, Erika Valeria Saliba;Penha Tinoco, Maria Laine;Miranda Brasileiro, Ana Cristina;Lima Aragao, Francisco Jose
    • Journal of Plant Biotechnology
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    • v.4 no.2
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    • pp.71-76
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    • 2002
  • In order to achieve repetitive somatic embryogenesis in cacao (Theobroma cacao L.), callus derived from floral tissues were continuously cultured in a medium containing 2,4-D. In 5% of the explants, repetitive somatic embryogenesis was observed after 8 weeks and maintained in a globular stage for several weeks. This is the first report showing repetitive somatic embryogenesis in cacao. The calli were bombarded with a plasmid containing $\beta$-glucuronidase (gus) as reporter gene. Two week old calli showed the high average number of cells expressing the us gene. The effect of osmotic agents (mannitol, sorbitol and sucrose) on gene expression was evaluated. Pre-treatment during 16 h with 0.25 M mannitol revealed an improvement in gene expression. The potential utilization of the repetitive embryogenesis, combined with osmotic treatment, is discussed as an alternative to achieve stable transgenic cacao plants.

Anti-Inflammatory Effects of Rice Bran Ethanol Extract in Murine Macrophage RAW 264.7 Cells (미강에탄올추출물의 RAW264.7 세포에서 항염증효과)

  • Park, Jeong-Suk;Kim, Mi-Hye
    • YAKHAK HOEJI
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    • v.55 no.6
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    • pp.456-461
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    • 2011
  • The aim of the present study is to investigate the anti-inflammatory effect of a Rice Bran Ethanol Extract (RBE). Inflammation, such as a bacterial infection in vivo metabolites, such as external stimuli or internal stimuli to the defense mechanisms of the biological tissue a variety of intracellular regulatory factors deulin inflammatory TNF-${\alpha}$, IL-$1{\beta}$, IL-6, IL-8, such as proinflammatory cytokines, prostagrandin, lysosomal enzyme, free radicals are involved in a variety of mediators. The present study was designed to determine the effect of the RBE on pro-inflammatory factors such as NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 in lipopolysaccharide (LPS) - stimulated RAW264.7 macrophages cells. The cell toxicity was determined by MTS assay. To evaluate of anti-inflammatory effect of RBE, amount of NO was measured using the NO detection kit and the iNOS expression was measured by reverse transcriptase polymerase chain reaction (RT-PCR). And proinflammatory cytokines were measured by ELISA kit. As a result, the RBE reduced NO, iNOS expression and TNF-${\alpha}$, IL-$1{\beta}$, IL-6 production without cytotoxicity. Our results suggest that the RBE may have an anti-inflammatory property through suppressing inflammatory mediator productions and appears to be useful as an anti-inflammatory material.

A Study on The of Prototype Representation (프로토타입 표현에 관한 연구)

  • Ock, Young-Jong;Yeo, Jeong-Mo
    • Proceedings of the Korean Institute of Information and Commucation Sciences Conference
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    • 2011.10a
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    • pp.721-724
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    • 2011
  • A prototype needs to show the requirements of the users and help them understand better. These prototypes are useful in a variety of industries. But most existing prototypes depend on the visual representation. This method of visual expression has a limitation, which can't express the detailed contents. This paper suggests an expression method of linguistic formation as an alternative. The prototype of linguistic expression can demonstrate the detailed contents of the requirements, keep a good communication among those who are related to the project, and allow them to deal with a large number of possible problems which may happen from the developing process. As it's also easy to make the developing process documented, the prototype will be helpful to improve the software development.

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Recombinant human BMP-2/-7 heterodimer protein expression for bone tissue engineering using recombinant baculovirus expression system

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong Ryul;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.32 no.2
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    • pp.49-53
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    • 2016
  • Bone morphogenetic proteins (BMPs) are essential growth factors for bone formation, skeletal development and bone regeneration. The BMP-2/7 heterodimer is known to have remarkable effects on osteogenic induction that are even stronger than the BMP-2 or BMP-7 homodimers. We designed a recombinant human BMP-2/7 (rhBMP-2/7) heterodimer protein with four glycine residues between BMP-2 and BMP-7 protein to facilitate free bond rotation of domains. The Baculovirus Expression Vector System (BEVS) is routinely used to produce recombinant proteins in the milligram scale. In this study, the BEVS was used to express the rhBMP-2/7 protein whrer the recombinant baculovirus was recovered in the host Sf9 cells. To confirm the biological activity of rhBMP-2/7 protein secreted from the BEVS as an osteogenic differentiation and induction factor, we measured the BMP-induced ALP activity. rhBMP-2/7 could be used as an alternative to BMPs to overcome limitations like short half-life and requirement for high concentrations. Furthermore, rhBMP-2/7 may be an efficient tool for various application studies such as bone regeneration and skeletal development.

A splice variant of human Bmal1 acts as a negative regulator of the molecular circadian clock

  • Lee, Jiwon;Park, Eonyoung;Kim, Ga Hye;Kwon, Ilmin;Kim, Kyungjin
    • Experimental and Molecular Medicine
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    • v.50 no.12
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    • pp.6.1-6.10
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    • 2018
  • Bmal1 is one of the key molecules that controls the mammalian molecular clock. In humans, two isoforms of Bmal1 are generated by alternative RNA splicing. Unlike the extensively studied hBmal1b, the canonical form of Bmal1 in most species, the expression and/or function of another human-specific isoform, hBmal1a, are poorly understood. Due to the lack of the N-terminal nuclear localization signal (NLS), hBMAL1a does not enter the nucleus as hBMAL1b does. However, despite the lack of the NLS, hBMAL1a still dimerizes with either hCLOCK or hBMAL1b and thereby promotes cytoplasmic retention or protein degradation, respectively. Consequently, hBMAL1a interferes with hCLOCK:hBMAL1b-induced transcriptional activation and the circadian oscillation of Period2. Moreover, when the expression of endogenous hBmal1a is aborted by CRISPR/Cas9-mediated knockout, the rhythmic expression of hPer2 and hBmal1b is restored in cultured HeLa cells. Together, these results suggest a role for hBMAL1a as a negative regulator of the mammalian molecular clock.