• BMB Reports
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• 제43권1호
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• pp.40-45
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• 2010

AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme that plays a central role in cellular metabolic stress. Modulation of nitric oxide (NO) and cyclooxygenase-2 (COX-2) is considered a promising approach for the treatment of inflammation and neuronal diseases. In this study, the AMPK gene was fused in-frame with PEP-1 peptide in a bacterial expression vector to produce a PEP-1-AMPK fusion protein. Expressed and purified PEP-1-AMPK fusion proteins were transduced efficiently into macrophage Raw 264.7 cells in a time- and dose-dependent manner. Furthermore, transduced PEP-1-AMPK fusion protein markedly inhibited LPS-induced iNOS and COX-2 expression. These results suggest that the PEP-1-AMPK fusion protein can be used for the protein therapy of COX-2 and NO-related disorders such as inflammation and neuronal diseases.

• Journal of Microbiology
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• 제43권3호
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• pp.285-294
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• 2005

A bacterial strain M4-7 capable of degrading various polyesters, such as poly$(\varepsilon-caprolactone)$, poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3-hydroxyoctanoate), and poly(3-hydroxy-5-phenylvalerate), was isolated from a marine environment and identified as Pseudomonas alcaligenes. The relative molecular mass of a purified extracellular medium-chain-length poly(3-hydroxyalkanoate) (MCL-PHA) depolymerase $(PhaZ_{palM4-7})$ from P. alcaligenes M4-7 was 28.0 kDa, as determined by SDS-PAGE. The $PhaZ_{palM4-7}$ was most active in 50 mM glycine-NaOH buffer (pH 9.0) at $35^{\circ}C$. It was insensitive to dithiothreitol, sodium azide, and iodoacetamide, but susceptible to p-hydroxymercuribenzoic acid, N-bromosuccinimide, acetic anhydride, EDTA, diisopropyl fluorophosphate, phenylmethylsulfonyl fluoride, Tween 80, and Triton X-100. In this study, the genes encoding MCL-PHA depolymerase were cloned, sequenced, and characterized from a soil bacterium, P. alcaligenes LB19 (Kim et al., 2002, Biomacro-molecules 3, 291-296) as well as P. alcaligenes M4-7. The structural gene $(phaZ_{palLB19})$ of MCL-PHA depolymerase of P. alcaligenes LB19 consisted of an 837 bp open reading frame (ORF) encoding a protein of 278 amino acids with a deduced $M_r$ of 30,188 Da. However, the MCL-PHA depolymerase gene $(phaZ_{palM4-7})$ of P. alcaligenes M4-7 was composed of an 834 bp ORF encoding a protein of 277 amino acids with a deduced Mr of 30,323 Da. Amino acid sequence analyses showed that, in the two different polypeptides, a substrate-binding domain and a catalytic domain are located in the N-terminus and in the C-terminus, respectively. The $PhaZ_{palLB19}$ and the $PhaZ_{palM4-7}$ commonly share the lipase box, GISSG, in their catalytic domains, and utilize $^{111}Asn$ and $^{110}Ser$ residues, respectively, as oxyanions that play an important role in transition-state stabilization of hydrolytic reactions.

• 만화애니메이션 연구
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• 통권45호
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• pp.417-434
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• 2016

알타미라 동굴의 다리 여덟 달린 멧돼지 벽화는 움직임을 표현하고자 하는 인류의 오랜 욕망을 보여주는 예로 제시되곤 한다. 그러나 여덟 개의 다리는 서로 다른 시기에 그려진, 이른바 더블 레이어의 결과물이다. 그럼에도 불구하고 알타미라 동굴벽화를 애니메이션의 시원과 결부시키는 설명은 별다른 의문이나 검증, 출처의 인용 등이 없이 꾸준히 재생산되고 있다. 여러 개의 다리를 움직임과 연결시키는 것은 시대를 초월한 시지각과는 무관하다. 그것은 특정한 시기에 발명된 움직임 표현 코드로 보아야 한다. 여기에는 19세기 후반에 시도된 연속촬영술, 그 중에서도 에티엔 쥘 마레의 시간사진술이 결정적인 역할을 한다. 일련의 연속적인 움직임들을 하나의 프레임 속에 겹쳐서 인화한 마레의 사진술은 20세기 초에 뒤샹과 발라를 비롯한 아티스트들의 회화 작업에 반영되면서 움직임 표현 코드로 형성되었다. 그 무렵부터 등장하기 시작한 애니메이션 매뉴얼들도 움직임을 분석하고 구현하기 위한 방법으로 마레의 시간사진술 이미지를 도입하였다. 결국 다리 여덟 개의 알타미라 동굴벽화를 움직임의 표현이라고 이해하는 것은 오늘날의 시각 코드를 통해 과거를 바라보고자 하는 오류이다.

• 대한바이러스학회지
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• 제30권1호
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• pp.71-81
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• 2000

We have determined and analyzed the full-length cDNA sequence of a coxsackievirus B3 (CVB3) Korean isolate (CVB3-Korea/97) which has been known as a general human pathogen. The whole genome contains 7,400 nucleotides and has a single large open reading frame with 6,555 nucleotides that encodes a potential polyprotein precursor of 2,185 amino acids. The genome also contains a 5' non-coding region (NCR) of 741 bases and a 3' NCR of 104 bases followed by poly(A) tail. Sequence homologies of nucleotides and deduced amino acids between the CVB3-Korea/97 strain and the prototype (Nancy strain) were 81.7% and 91.5%, respectively. The genes encoding the functional proteins including viral protease and RNA dependent RNA polymerase showed higher homology than those encoding the structural proteins. We have further analyzed the sequences of 5' NCR, VP1 and VP2 of CVB3-Korea/97, which are known as cardiovirulent determining factors at the nucleotide and amino acid levels. Although the CVB 3-Korea/97 strain was isolated from an aseptic meningitis patient without cardiomyopathy, its 234th nucleotide and 165th amino acid were uracil and Asn as same as those of other cardiovirulent strains one. However, the 155th amino acid of VP1, which closely associated with cardiovirulence, was replaced with $Arg^{155}$ by single nucleotide substitution from $A^{2916}$ to $T^{2916}$. Moreover, additional amino acid substitutions were observed in the flanking region of $Asp^{155}$. Taken together, amino acid(s) substitution in VP1 may playa critical role in determining cardiovirulence of the CVB3-Korea/97 strain rather than individual nucleotide replacements in the 5' NCR and/or an amino acid substitution in VP2.

• 생명과학회지
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• 제24권12호
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• pp.1291-1300
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• 2014

본 연구에서는 참전복(Haliotis discus hannai)의 대용량 염기서열 분석을 통해 peroxiredoxin (Prx) 2 유전자의 full length의 cDNA를 동정하였다. 참전복 Prx 2 유전자의 총 길이는 1,052 bp로 597 bp의 open reading frame는 총 199개의 아미노산을 코딩하고 있으며 분자량은 22 kDa, 등전점은 7.58로 예측되었다. 참전복 Prx 2 아미노산서 열에는 typical 2-Cys Prx의 특징을 갖는 motif와 효소활성에 중요한 cysteine잔기가 매우 보존되어 있었다. 참전복 Prx 2 아미노산 서열은 다른 종의 Prx 2와 64~99% 유사하였고, 특히 패류의 Prx 2와 가장 유사성이 높았다. 참전복 Prx 2 유전자의 mRNA는 관찰된 모든 조직에서 발현하고 있었으며, 특히 외투막, 아가미, 족부, 간췌장, 소화관에서 높은 발현이 확인되었다. 참전복의 Prx 2는 비브리오균으로 감염시킨 전복의 혈구세포에서 감염 후 1시간 째 발현이 증가했다가 서서히 감소하였고, 간췌장 조직에서는 감염 6시간 경과 후 발현 정도가 최고로 증가했다가 감소하였다. 따라서 typical 2-Cys Prx의 특징을 잘 보존되어 있는 참전복 Prx 2는 병원균 감염에 따른 산화스트레스 조절에 관여하는 인자로 중요한 역할을 할 것으로 예상된다.

• Plant Biotechnology Reports
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• 제3권2호
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• pp.147-155
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• 2009

Oryza grandiglumis Chitinase IVa (OgChitIVa) cDNA encoding a class IV chitinase was cloned from wild rice (Oryza grandiglumis). OgChitIVa cDNA contains an open reading frame of 867 nucleotides encoding 288 amino acid residues with a predicted molecular weight of 30.4 kDa and isoelectric point of 8.48. Deduced amino acid sequences of OgChitIVa include the signal peptide and chitin-binding domain in the N-terminal domain and conserved catalytic domain. OgChitIVa showed significant similarity at the amino acid level with related monocotyledonous rice and maize chitinase, but low similarity with dicotyledoneous chitinase. Southern blot analysis showed that OgChitIVa genes are present as two copies in the wild rice genome. It was shown that RNA expression of OgChitIVa was induced by defense/stress signaling chemicals, such as jasmonic acid, salicylic acid, and ethephon or cantharidin and endothall or wounding, and yeast extract. It was demonstrated that overexpression of OgChitIVa in Arabidopsis resulted in mild resistance against the fungal pathogen, Botrytis cinerea, by lowering disease rate and necrosis size. RT-PCR analysis showed that PR-1 and PR-2 RNA expression was induced in the transgenic lines. Here, we suggest that a novel OgChitIVa gene may play a role in signal transduction process in defense response against B. cinerea in plants.

• Journal of Animal Science and Technology
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• 제50권2호
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• pp.265-272
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• 2008

지방세포분화 초기에 중요한 역할을 수행하는 것으로 보고되고 있는 C/EBPβ 유전자를 cloning하기 위하여 한우 26개월령 등심조직을 이용하여 Total RNA를 추출하고 RT-PCR을 수행한 결과 한우 및 흑한우 C/EBPβ는 1047bp, 348 아미노산 서열을 가지고 있었다. 한우 및 흑한우 C/EBPβ의 아미노산 서열을 NCBI에 등록된 Japanese black cattle C/EBPβ(NCBI accession No. NM_176788) 비교한 결과 약 97%의 상동성을 나타내었다. 또한 인간, 닭, 생쥐, 쥐의 C/EBPβ 아미노산 서열을 비교한 결과 각각 96%, 58%, 74%, 75%의 상동성을 나타내었다. 그러나 각 종간의 leucine zipper domain간의 상동성은 매우 높게 나타났다. 한우 C/EBPβ의 발현은 폐, 등심, 지방조직에서 나타나고 있으나, 지방조직에서 좀 더 많은 발현을 보이고 있었다. 그리고 C/EBPβ 유전자의 bZIP 영역에서 polymorphism이 나타나지 않음을 확인하였다.

• 정보관리연구
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• 제35권1호
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• pp.1-27
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• 2004

지식경영에 있어서 조직 구성원들 간에 지식을 공유하는 가장 기초적이면서 기틀이 되는 것은 공유할 지식을 체계적으로 관리할 수 있는 지식분류체계를 구축하는 것이다. 이에 이 연구에서는 지식경영의 도입단계에 있는 한국원자력연구소에서 조직 구성원들이 실제로 지식을 효율적으로 공유할 수 있도록 하기 위한 최적의 지식분류체계를 구축하였다. 이 연구에서는 지식경영의 본격적인 도입에 따라 한국원자력연구소에서 실제로 활용할 수 있는 전소차원의 지식분류체계를 구축하기 위해서, 기본적인 분류체계 설계원칙 아래 기존의 다양한 분류체계를 참고하여, (1) 지식분류체계 초안 작성단계, (2) 지식분류체계 수정안 작성단계, (3) 지식분류체계 검증 및 확정단계 등과 같이 3단계 과정을 거쳐 지식분류체계를 완성하였다. 완성된 지식분류체계는 기술적인 측면과 조직체계를 감안한 미래지향적인 분류체계로서 총 218항목으로서, 대분류 8항목, 중분류 43항목, 소분류 167항으로 구성되어 있다. 여기에서 설계된 지식분류체계는 한국원자력연구소에서 지식경영을 운영할 때 지식의 체계적인 관리를 통한 조직 구성원들의 지식 공유 및 활용의 가장 기초적이면서 중요한 틀로써 이용될 수 있을 것이다. 아울러 지식분류체계 자체는 물론 지식분류체계의 초안 작성에서부터 최종안을 완성하기까지의 제반 설계과정 및 방법론적인 측면은 지식경영을 도입하려는 타 연구기관이나 기업체 등에서 지식분류체계를 설계할 때 매우 유용하게 활용될 수 있을 것으로본다.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2000년도 International Meeting 2000
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• pp.23-36
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• 2000

Japanese encephalitis virus (JEV) is the causative agent of a mosquito-borne encephalitis and is transmitted to human via persistently infected mosquito vectors. Although the virus is known to cause only acute infection, there were reports that showed neurological sequelae, latent infection in peripheral mononuclear cells, and recurrence of the disease after acute encephalitis. Innate resistance of certain cell lines, abnormal SN1 expression of the virus, and anti-apoptotic effect of cullular bcl-2 have been suggested as probable causes of JEV persistence even in the absence of defective interfering (DI) particles. Although possible involvement of DI particles in JEV persistence was suggested, neither has a direct evidence for DI presence nor its molecular characterization been made. Two questions asked in this study are whether the DI virus plays any role in JEV persistent infection if it is associated with and what type of change(s) can be made in persistently infected cells to avoid apoptosis even with the continuous virus replication, DI-free standard stock of JEV was infected in BHK-21, Vero, and SW13 cells and serial high multiplicity passages were performed in order to generate DI particles. There different-sized DI RNA species which were defective in both structural and nonstructural protein coding genes. Rescued ORFs of the DI genome maintained in-frame and the presence of replicative intermediate or replicative form RNA of the DI particles confirmed their replication competence. On the other hand, several clones with JEV persistent infection were established from the cells survived acute infections during the passages. Timing of the DI virus generation during the passages seemed coincide to the appearance of persistently infected cells. The DI RNAs were identified in most of persistently infected cells and were observed throughout the cell maintenance. One of the cloned cell line maintained the viral persistence without DI RNA coreplication. The cells with viral persistence released the reduced but continuous infectious JEV particle for up to 9 months and were refractory to homologous virus superinfection but not to heterologous challenges. Unlike the cells with acute infection these cells were devoid of characteristic DNA fragmentation and JEV-induced apoptosis with or without homologous superinfection. Therefore, the DI RNA generated during JEV undiluted serial passage on mammalian cells was shown to be biologically active and it seemed to be responsible, at least in part, for the establishment and maintenance of the JEV persistence in mammalian cells. Viral persistence without DI RNA coreplication, as in one of the cell clones, supports that JEV persistent infection could be maintained with or without the presence of DI particles. In addition, the fact that the cells with JEV persistence were resistant against homologous virus superinfection, but not against heterologous one, suggests that different viruses have their own and independent pathway for cytopathogenesis even if viral cytopathic effect could be converged to an apoptosis after all.