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Cloning and Characterization of Monofunctional Catalase from Photosynthetic Bacterium Rhodospirillum rubrum S1

  • Lee, Dong-Heon;Oh, Duck-Chul;Oh, You-Sung;Malinverni, Juliana C.;Kukor, Jerome J.;Kahng, Hyung-Yeel
    • Journal of Microbiology and Biotechnology
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    • v.17 no.9
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    • pp.1460-1468
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    • 2007
  • In this study, an approx. 2.5-kb gene fragment including the catalase gene from Rhodospirillum rubrum S1 was cloned and characterized. The determination of the complete nucleotide sequence revealed that the cloned DNA fragment was organized into three open reading frames, designated as ORF1, catalase, and ORF3 in that order. The catalase gene consisted of 1,455 nucleotides and 484 amino acids, including the initiation and stop codons, and was located 326 bp upstream in the opposite direction of ORF1. The catalase was overproduced in Escherichia coli UM255, a catalase-deficient mutant, and then purified for the biochemical characterization of the enzyme. The purified catalase had an estimated molecular mass of 189 kDa, consisting of four identical subunits of 61 kDa. The enzyme exhibited activity over a broad pH range from pH 5.0 to pH 11.0 and temperature range from $20^{\circ}C$ to $60^{\circ}C$C. The catalase activity was inhibited by 3-amino-1,2,4-triazole, cyanide, azide, and hydroxylamine. The enzyme's $K_m$ value and $V_{max}$ of the catalase for $H_2O_2$ were 21.8 mM and 39,960 U/mg, respectively. Spectrophotometric analysis revealed that the ratio of $A_{406}$ to $A_{280}$ for the catalase was 0.97, indicating the presence of a ferric component. The absorption spectrum of catalase-4 exhibited a Soret band at 406 nm, which is typical of a heme-containing catalase. Treatment of the enzyme with dithionite did not alter the spectral shape and revealed no peroxidase activity. The combined results of the gene sequence and biochemical characterization proved that the catalase cloned from strain S1 in this study was a typical monofunctional catalase, which differed from the other types of catalases found in strain S1.

ORF5a Protein of Porcine Reproductive and Respiratory Syndrome Virus is Indispensable for Virus Replication (PRRS 바이러스 ORF5a 단백질의기능학적역할)

  • Oh, Jongsuk;Lee, Changhee
    • Microbiology and Biotechnology Letters
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    • v.43 no.1
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    • pp.1-8
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    • 2015
  • In this study, a DNA-launched reverse genetics system was developed from a type 2 porcine reproductive and respiratory syndrome virus (PRRSV) strain, KNU-12. The complete genome of 15,412 nucleotides was assembled as a single cDNA clone and placed under the eukaryotic CMV promoter. Upon transfection of BHK-tailless pCD163 cells with a full-length cDNA clone, viable and infectious type 2 progeny PRRSV were rescued. The reconstituted virus was found to maintain growth properties similar to those of the parental virus in porcine alveolar macrophage (PAM) cells. With the availability of this type 2 PRRSV infectious clone, we first explored the biological relevance of ORF5a in the PRRSV replication cycle. Therefore, we used a PRRSV reverse genetics system to generate an ORF5a knockout mutant clone by changing the ORF5a translation start codon and introducing a stop codon at the 7th codon of ORF5a. The ORF5a knockout mutant was found to exhibit a lack of infectivity in both BHK-tailless pCD163 and PAM-pCD163 cells, suggesting that inactivation of ORF5a expression is lethal for infectious virus production. In order to restore the ORF5a gene-deleted PRRSV, complementing cell lines were established to stably express the ORF5a protein of PRRSV. ORF5a-expressing cells were capable of supporting the production of the replicationdefective virus, indicating complementation of the impaired ORF5a gene function of PRRSV in trans.

A study on the uniqueness of 's hyper-realistic animation style (영화 <아바타>가 보여주는 극사실적 애니메이션 스타일의 특이성 연구)

  • Lee, Youn-H.
    • Cartoon and Animation Studies
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    • s.20
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    • pp.47-61
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    • 2010
  • The Uncanny Valley is a hypothesis introduced by a roboticist, Masahiro Mori. The theory holds that as robots and other facsimiles of humans look and act more human-like, the emotional response from a human spectator to them will become increasingly empathic, until they approach realistic similarity to humans but not quite exactly like them, when those images stop being likable and instead become repulsive and "uncanny." Although the scientific validity of this hypothesis is still debatable, it is true that many spectators has tendency to reject hyper-realistic animation. For that reason, the great success of the recent movie Avatar is remarkable. Computer animations that try to represent humans in hyper-realistic way fall into a paradox: successful result images would look too mundane and unsuccessful ones look "wrong". On top of that, audience would expect from hyper-realistic animation to see same level of acting of live-action films, which is usually not the case. The subtle differences between digital characters and real actors irritate spectators and hinder them in absorbtion. The biggest difference between Avatar and other hyper-realistic animation is the referent of their representation. While Avatar pursues a similar hyper-realistic visual style, the referents of the representation shown in this movie are totally alien to us thus impossible to compare with the real counterparts. Unlike characters in other hyper-realistic animations who had to fight an uphill battle with great actors in real life, those in Avatar have criteria of their own. That is why this movie can be free from the dreadful valley and let the audience absorb into the spectacle.

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Study on the energy-saving constant temperature and humidity machine operating characteristics (에너지 절감형 항온항습기 운전 특성에 관한 연구)

  • Cha, Insu;Ha, Minho;Jung, Gyeonghwan
    • Journal of Energy Engineering
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    • v.25 no.3
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    • pp.27-33
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    • 2016
  • The heat recovery system that was applied in this study, is the energy-saving type that can produce the maximum cooling capacity less power in use. In order to have a more precise control function the temperature and humidity of the constant temperature and humidity machine, control algorithm is applied to designed a fuzzy PID controller, and the outside air compensation device (air-cooled) demonstrated excellent ability to dehumidify the moisture, $-20^{\circ}C$ in winter. High efficiency and the low-noise type sirocco fan operate quitely and designed to fit the bottom-up and top-down in accordance with the characteristics of equipment. as a result of experiment data, the conversion efficiency is 95% or more, power recovery time is within 5sec, stop delay time is within 30sec, pump down time is 10sec, pump delay time is 5sec, heating delay time is 5sec, temperature deviation is ${\pm}2^{\circ}C$ (cooling deviation: $2^{\circ}C$, Heating deviation : $2^{\circ}C$), humidity deviation is a ${\pm}5%$ (humidification deviation 3.0%, dehumidification deviation 3.0%). Recently, ubiquitous technology is important. so, the constant temperature and humidity machine designed to be able to remotely control to via the mobile phone, and more scalable to support MMI software and automatic interface. Further, the life of the parts and equipment is extended by the failure.

The Effects of NogYongDaeBoTang,(NYDBT)on the Alzheimer's Disease Model Induced by CT-105 and $A{\beta}$ (녹용대보탕이 ${\beta}-Amyloid$로 유도(誘導)된 Alzheimer's Disease 병태(病態) 모델에 미치는 영향(影響))

  • Seo, Gyoo-Tae;Lee, Eun-Kyung;Choi, Cheol-Hong;Chung, Dae-Kyoo
    • Journal of Oriental Neuropsychiatry
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    • v.18 no.2
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    • pp.101-132
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    • 2007
  • Objective : This research investigates the effect of the NogYongDaeBoTang,(NYDBT) on Alzheimer's disease. Method : The effects of the NYDBT extract on (1) $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ mRNA of PC-12 cells treated with LPS; (2) acetylcholinesterase(AChE), amyloid precursor proteins(APP), and glial fibrillary acidic protein(GFAP) mRNA the AChE activity and the APP production of PC-12 cell treated with CT-105; (3) the behavior; (4) expression of $IL-1{\beta}$, $TNF-{\alpha}$, MDA, $IL-1{\beta}$ mRNA, and $TNF-{\alpha}$ mRNA; (5) the infarction area of the hippocampus, and brain tissue injury in Alzheimer‘s diseased mice induced with ${\beta}A$ were investigated. Results : 1. The NYDBT extract suppressed the expression of $IL-1{\beta}$, IL-6 and $TNF-{\alpha}$ mRNA in BV2 microglia cell line treated with LPS. 2. The NYDBT extract suppressed the expression of $IL-1{\beta}$, IL-6, and $TNF-{\alpha}$ protein production in BV2 microglia cell line treated with LPS. 3. For the NYDBT extract group a significant inhibitory effect on the memory deficit was shown for the mice with Alzheimer's disease induced by $A{\beta}$ in the Morris water maze experiment, which measured stop-through latency, and distance movement-through latency. 4. The NYDBT extract suppressed the over-expression of $IL-1{\beta}$ protein, $TNF-{\alpha}$ protein, MDA, and CD68/CD11b, in the mice with Alzheimer's disease induced by $A{\beta}$. 5. The NYDBT extract reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by $A{\beta}$. 6. The NYDBT extract reduced the Tau protein, GFAP protein, and presenilin1/2 protein (immunohistochemistry) of hippocampus in the mice with Alzheimer's disease induced by $A{\beta}$. Conclusions : These results suggest that the NYDBT extract may be effective for the prevention and treatment of Alzheimer's disease.

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The Effects of Hominis Placenta Herbal-Acupuncture Solution on the Alzheimer's Disease Model Induced by ${\beta}A$ (자하차(紫河車) 약침(藥鍼)이 ${\beta}A$로 유도(誘導)된 Alzheimer's Disease 병태(病態) 모델에 미치는 영향(影響))

  • Lee, Byung-Hun;Park, Sun-Young;Choi, Cheol-Hong;Lee, Eun-Kyung;Chung, Dae-Kyoo
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.2
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    • pp.41-64
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    • 2008
  • Objective: Hominis Placenta is used in many cure, mainly treats a weak, chronic disease, especially senile. This research investigates the effect of the Hominis Placenta Herbal-Acupuncture Solution on Alzheimer's disease. Method: The effects of the Hominis Placenta Herbal-Acupuncture Solution on (1) $IL-1{\beta}$ protein, $TNF-{\alpha}$ protein, MDA, and CD68/CD11b (2) the behavior (3) the infarction area of the hippocampus, and brain tissue injury in Alzheimer's diseased mice induced with 13A were investigated. Results: 1. For the Hominis Placenta Herbal-Acupuncture Solution group a significant inhibitory effect on the memory deficit was shown for the mice with Alzheimer's disease induced by ${\beta}$ A in the Morris water maze experiment, which measured stop-through latency, and distance movement-through latency. 2. The Hominis Placenta Herbal-Acupuncture Solution group suppressed the over-expression of $IL-1{\beta}$ protein, $TNF-{\alpha}$ protein, MDA, and CD68/CD11b, in the mice with Alzheimer's disease induced by ${\beta}A$. 3. The Hominis Placenta Herbal Acupuncture Solution group reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by ${\beta}A$. 4. The Hominis Placenta Herbal-Acupuncture Solution group reduced the Tau protein, GFAP protein, and presenilin1/2 protein, beta-secretase protein, (immunohistochemistry) of hippocampus in the mice with Alzheimer's disease induced by ${\beta}A$. Conclusion: These results suggest that the Hominis Placenta Herbal-Acupuncture Solution group may be effective for the prevention and treatment of Alzheimer's disease. Investigation into the clinical use of the Hominis Placenta Herbal-Acupuncture Solution for Alzheimer's disease is suggested for future research.

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Action mechanism of upstream open reading frame from S-adenosylmethionine decarboxylase gene as a in vivo translational inhibitor (S-Adenosylmethionine decarboxylase 유전자의 upstream open reading frame이 in vivo에서 translational inhibitor 로서의 작용 기작)

  • Choi, Yu-Jin;Park, Ky-Young
    • Journal of Plant Biotechnology
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    • v.38 no.1
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    • pp.87-93
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    • 2011
  • S-Adenosylmethionine decarboxylase (SAMDC; EC 4.1.4.50), a key enzyme for polyamines biosynthesis, was tightly regulated for homeostatic levels. Carnation SAMDC gene (CSDC9) has an small upstream open reading frame (uORF) of 54 amino acids in 5'-leader sequence. To explore the functional mechanism of uORFs in controlling translation, we used a GUS reporter gene driven with the 35S promoter and uORF region of SAMDC gene for making transgenic tobacco plants. In our experiment, there were a translational inhibition of its downstream GUS ORF by SAMDC uORF sequence or SAMDC uORF protein. Expecially, translational inhibition was most effective in point-mutated construct, in which the start codon was changed. Therefore, this results suggested the ribosomal stalling might be involved in this translational inhibitory process. The frame shift in amino acid sequence of SAMDC uORF with start codon and stop codon resulted in a moderate increasing in GUS activity, suggesting the native amino acid sequence was important for a function as a translational inhibitor. Also, we showed that the production of GUS protein was significantly inhibited in the presence of the small uORF using histochemical analysis of GUS expression in seedlings and tobacco flowers. Importantly, the small uORF sequence induced a real peptide of 5.7 kDa, which was provided the presence of SAMDC uORF peptide band using an in vitro transcription/translation system. The peptide product of uORF might interact with other components of translational machinery as well as polyamines, which was resulted from that polyamine treatment was inhibited GUS protein band in SDS-PAGE experiment.

Site Application Characteristics of Deep-Site Biopile System for Cleaning Oil-Contaminated Soil/Underground Water (유류오염 토양/지하수 정화를 위해 개발된 DSB(Deep-Site Biopile) System 현장적용특성)

  • Han Seung-Ho;Kong Sung-Ho;Kang Jung-Woo
    • Journal of Soil and Groundwater Environment
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    • v.10 no.2
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    • pp.28-34
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    • 2005
  • The aim of this article is to assess the application characteristics of the site by remediating oil-contaminated area using DSB (Deep-site Biopile) system. In the contaminated area, the soil was composed of penetrable sand and the leaked oil was spread widely (total 7,201 cubic meters) through 2.5 meter deep underground water flow. DSB system was operated for 30 minutes intervals for 24 hours in a day (30 minutes opεration and 30 minutes stop). To check contamination level change in the contaminated area after DSB system was operated, samples were taken. The result from the site shows that BTEX/TPH contamination level was dropped 50% after 30-day operation of DSB system, and that contamination level was dropped below contamination level check standard after 165 days and the remediation was completed. Unlike traditional biological remediation methods DSB system could efficiently process soil and water which were contaminated by high levels of oil compounds.

Design and implementation of a 3-axis Motion Sensor based SWAT Hand-signal Motion-recognition System (3축 모션 센서 기반 SWAT 수신호 모션 인식 시스템 설계 및 구현)

  • Yun, June;Pyun, Kihyun
    • Journal of Internet Computing and Services
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    • v.15 no.4
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    • pp.33-42
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    • 2014
  • Hand-signal is an effective communication means in the situation where voice cannot be used for expression especially for soldiers. Vision-based approaches using cameras as input devices are widely suggested in the literature. However, these approaches are not suitable for soldiers that have unseen visions in many cases. in addition, existing special-glove approaches utilize the information of fingers only. Thus, they are still lack for soldiers' hand-signal recognition that involves not only finger motions, but also additional information such as the rotation of a hand. In this paper, we have designed and implemented a new recognition system for six military hand-signal motions, i. e., 'ready', 'move', quick move', 'crawl', 'stop', and 'lying-down'. For this purpose, we have proposed a finger-recognition method and motion-recognition methods. The finger-recognition method discriminate how much each finger is bended, i. e., 'completely flattened', 'slightly flattened', 'slightly bended', and 'completely bended'. The motion-recognition algorithms are based on the characterization of each hand-signal motion in terms of the three axes. Through repetitive experiments, our system have shown 91.2% of correct recognition.

Characterization of Mud Loach (Misgurnus mizolepis) Apolipoprotein A-I: cDNA Cloning, Molecular Phylogeny and Expression Analysis (미꾸라지(Misgurnus mizolepis) Apolipoprotein A-I cDNA의 구조, 분자계통 및 발현 특징 분석)

  • Lee, Youn-Ho;Noh, Jae-Koo;Kim, Keun-Yong;Cho, Young-Sun;Nam, Yoon-Kwon;Kim, Dong-Soo
    • Journal of Aquaculture
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    • v.20 no.1
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    • pp.65-72
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    • 2007
  • Full length complementary DNA encoding apolipoprotein A-I (apoA-I) was isolated and characterized in mud loach (Misgurnus mizolepis). Mud loach apoA-I cDNA encoding 24 bp of 5'-untranslated region (UTR), 762 bp of single open reading frame (ORF) consists of 254 amino acids and 293 bp of 3'-UTR excluding stop codon and poly (A+) tail. Two overlapping polyadenylation signals (AATAAAATAAA) was found 9 bp prior to the poly (A+) tail. Mud loach apoA-I represented considerable homology to those from other teleost species at amino acid level with conserving common features of vertebrate apoA-I. Molecular phylogenetic analysis inferred the phylogenetic hypothesis that was generally in accordance with the previous taxonomic relationship. Apolipoprotein A-I mRNA was detected in various tissues, but the mRNA levels were quite varied depending on tissues based on semi-quantitative RT-PCR. Liver and brain showed the significantly higher levels of apoA-I transcripts than other tissues. mRNA expression of apoA-I was quite low in very early stage of embryonic development, however dramatically enhanced from 8 hours post fertilization. This increased mRNA level was retained consistently up to 14 days post hatching.