• 제목/요약/키워드: CDS1

검색결과 159건 처리시간 0.023초

Cloning and Expression of the Cathepsin F-like Cysteine Protease Gene in Escherichia coli and Its Characterization

  • Joo, Han-Seung;Koo, Kwang-Bon;Park, Kyun-In;Bae, Song-Hwan;Yun, Jong-Won;Chang, Chung-Soon;Choi, Jang-Won
    • Journal of Microbiology
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    • 제45권2호
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    • pp.158-167
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    • 2007
  • In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

폴리페놀계 천연 항산화제의 cyclodextrin inclusion complexation을 통한 안정화와 식품 보존제로의 활용 (Stabilization of Polyphenolic Antioxidants Using Inclusion Complexation with Cyclodextrin and Their Utilization as the Fresh-food Preservative)

  • 김태권;신현동;이용현
    • 한국식품과학회지
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    • 제35권2호
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    • pp.266-271
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    • 2003
  • CD의 inclusion complex 형성능을 이용하여 대표적인 polyphenol계 천연 항산화 물질인 quercertin 및 catechin을 안정화시키는 분자 캡슐화 기술을 연구하였다. 먼저 ${\alpha}-,\;{\beta}-$, 그리고 ${\gamma}-CD$의 quercetin 및 catechin의 포접능을 비교하였다. Quercetin/catechin-CD inclusion complex 형성을 DSC thermogram을 이용하여 확인하였다. 또한 ${\beta}-CD$와의 포접에 따른 저용해성 quercetin과 catechin의 물에 대한 용해도의 변화를 관찰한 결과 inclusion complex 형성 시 용해도가 증가하는 것을 관찰할 수 있었다. Inclusion complex의 온도 및 pH에 대한 안정성을 검토한 결과 inclusion complex가 원래의 물질보다 높은 항산화능을 유지하였으며 특히 catechin의 안정성이 크게 향상되었다. Catechin-CD inclusion complex의 식품보존제로서의 적합성을 지방산인 linoleic acid를 시료로 검토하기 위하여 저장 중 과산화물가의 변화를 측정한 결과 포접된 catechin이 보다 높은 항산화능을 보였다. 이와 같은 용해도 및 항산화능의 증가로 볼 때 quercetin/catechin-CD inclusion complex는 신선식품의 보존제로 효과적으로 활용될 수 있는 것으로 판단된다.

Characterization, Cloning and Expression of the Ferritin Gene from the Korean Polychaete, Periserrula leucophryna

  • Jeong Byeong Ryong;Chung Su-Mi;Baek Nam Joo;Koo Kwang Bon;Baik Hyung Suk;Joo Han-Seung;Chang Chung-Soon;Choi Jang Won
    • Journal of Microbiology
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    • 제44권1호
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    • pp.54-63
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    • 2006
  • Ferritin is a major eukaryotic protein and in humans is the protein of iron storage. A partial gene fragment of ferritin (255 bp) taken from the total RNA of Periserrula leucophryna, was amplified by RT-PCR using oligonucleotide primers designed from the conserved metal binding domain of eukaryotic ferritin and confirmed by DNA sequencing. Using the $^{32}P-labeled$ partial ferritin cDNA fragment, 28 different clones were obtained by the screening of the P. leucophryna cDNA library prepared in the Uni-ZAP XR vector, sequenced and characterized. The longest clone was named the PLF (Periserrula leucophryna ferritin) gene and the nucleotide and amino acid sequences of this novel gene were deposited in the GenBank databases with accession numbers DQ207752 and ABA55730, respectively. The entire cDNA of PLF clone was 1109 bp (CDS: 129-653), including a coding nucleotide sequence of 525 bp, a 5' -untranslated region of 128 bp, and a 3'-noncoding region of 456 bp. The 5'-UTR contains a putative iron responsive element (IRE) sequence. Ferritin has an open reading frame encoding a polypeptide of 174 amino acids including a hydrophobic signal peptide of 17 amino acids. The predicted molecular weights of the immature and mature ferritin were calculated to be 20.3 kDa and 18.2 kDa, respectively. The region encoding the mature ferritin was subcloned into the pT7-7 expression vector after PCR amplification using the designed primers and included the initiation and termination codons; the recombinant clones were expressed in E. coli BL21(DE3) or E. coli BL21(DE3)pLysE. SDS-PAGE and western blot analysis showed that a ferritin of approximately 18 kDa (mature form) was produced and that by iron staining in native PAGE, it is likely that the recombinant ferritin is correctly folded and assembled into a homopolymer composed of a single subunit.

Mapping, Tissue Distribution and Polymorphism of Porcine Retinol Binding Protein Genes (RBP5 and RBP7)

  • Gong, W.H.;Tang, Z.L.;Han, J.L.;Yang, S.L.;Wang, H.;Li, Y.;Li, K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권11호
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    • pp.1544-1550
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    • 2008
  • The retinoids (vitamin A and its derivatives) play a critical role in vision, growth, reproduction, cell differentiation and embryonic development. Using the IMpRH panel, porcine cellular retinol binding protein genes 5 and 7 (RBP5 and RBP7) were assigned to porcine chromosomes 5 and 6, respectively. The complete coding sequences (CDS) of the RBP5 and RBP7 genes were amplified using the reverse transcriptase polymerase chain reaction (RT-PCR) method, and the deduced amino acid sequences of both genes were compared to human corresponding proteins. The mRNA distributions of the two genes in adult Wuzhishan pig tissues (lung, skeletal muscle, spleen, heart, stomach, large intestine, lymph node, small intestine, liver, brain, kidney and fat) were examined. A total of nine single nucleotide polymorphisms (SNPs) were identified in two genes. Three of these SNPs were analyzed using the polymerase chain reaction-restriction-fragment length polymorphism (PCR-RFLP) method in Laiwu, Wuzhishan, Guizhou, Bama, Tongcheng, Yorkshire and Landrace pig breeds. Association analysis of genotypes of these SNP loci with economic traits was done in our experimental populations. Significant associations of different genotypes of $RBP5-A/G^{63}$, $RBP5-A/G^{517}$ and $RPB5-T/C^{intron1-90}$ loci with traits including maximum carcass length (LM), minimum carcass length (LN), marbling score (MS), back fat thickness at shoulder (SBF), meat color score (MCS) and hematocrit (HCT) were detected. These SNPs may be useful as genetic markers in genetic improvement for porcine production.

인공위성 레이저추적 시스템(ARGO-M) 개발 현황 (STATUS AND PROGRESS OF ARGO-M SYSTEM DEVELOPMENT)

  • 박은서;유성열;임형철;방승철;서윤경;박장현;조중현;박종욱;나자경;장정균;장비호;김광동;김병인;박철훈;이성휘;함상용;손영수
    • 천문학논총
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    • 제27권3호
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    • pp.49-59
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    • 2012
  • KASI (Korea Astronomy and Space Science Institute) has developed an SLR (Satellite Laser Ranging) system since 2008. The name of the development program is ARGO (Accurate Ranging system for Geodetic Observation). ARGO has a wide range of applications in the satellite precise orbit determination and space geodesy research using SLR with mm-level accuracy. ARGO-M (Mobile, bistatic 10 cm transmitting/40 cm receiving telescopes) and ARGO-F (Fixed stationary, about 1 m transmitting/receiving integrated telescope) SLR systems development will be completed by 2014. In 2011, ARGO-M system integration was completed. At present ARGO-M is in the course of system calibration, functionality, and performance tests. It consists of six subsystems, OPS (Optics System), TMS (Tracking Mount System), OES (Opto-Electronic System), CDS (Container-Dome System), LAS (Laser System) and AOS (ARGO Operation System). In this paper, ARGO-M system structure and integration status are introduced and described.

알츠하이머병, 파킨슨병 및 혈관성치매 환자들의 우울증에 대한 약물 치료 평가 (Evaluation of Pharmacotherapy for the Patients with Depression in Alzheimer's Disease, Parkinson's Disease or Vascular Dementia)

  • 이효진;이옥상;정선회;박미숙;임성실
    • 한국임상약학회지
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    • 제23권1호
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    • pp.33-41
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    • 2013
  • Background: Prevalence of depression comorbid with neurologic disorders such as Alzheimer' disease (AD), Parkinson's disease (PD) and vascular dementia (VD) is higher than that of primary depression. Antidepressant medications, suggested by many researches for depression comorbid with neurologic disorders such as AD, PD and VD, are mainly selective serotonin reuptake inhibitors (SSRI). Objective: The primary objective of this study is the evaluation of antidepressant drug therapy for AD, PD and VD. Method: This study was a retrospective study based on medical records, carried out for 3 years and 6 months (Jan. 2007~Jul. 2010). Patients, diagnosed as comorbid depression through Beck Depression Inventory (BDI), Cornell Depression Scale (CDS), Geriatric Depression Scale (GDS) among neurologic out-patients of Chungnam National University Hospital because of AD, PD and VD, were selected. The results were evaluated by efficacy and safety of antidepressant drug therapy. Results: In result, the prescribing rates of antidepressants were 30%, 55% and 40% for each AD, PD and VD. Depression cure rates of patients receiving antidepressants vs patients not receiving antidepressants were 40% vs 39%, 33% vs 23% and 38% vs 30% for AD, PD and VD. The frequencies of prescriptoin of SSRI were 21%, 11% and 27% for each AD, PD and VD. The frequencies of prescriptoin of benzodiazepine (BZD) was 61%, 82% and 61% for each AD, PD and VD. The ratio of single BZD prescription was more than that of combination prescription of antidepressants. Tricyclic antidepressants (TCA) were rarely prescribed. The rate of patients with BZD-related side effects was 54%. The most frequent side effects of BZD were dizziness (30%), drowsiness (21%) and headache (16%). Side effects of SSRI were rare. Conclusion: In conclusion, the frequencies of prescription of antidepressants were not common for AD, PD and VD. There was little difference in depression cure rate between patient receiving antidepressants and not receiving. Even though SSRI has to be the highest priority of usage, the frequencies of prescription of SSRI were lower than those of BZD. Additional researches and efforts are required to improve antidepressant drug therapy for neurologic disorders such as AD, PD and VD.

해양 해면체로부터 분리한 세균으로 항알러지성물질을 생산하는 Bacillus safensis KCTC 12796BP의 유전체 해독 (The complete genome sequence of a marine sponge-associated bacteria, Bacillus safensis KCTC 12796BP, which produces the anti-allergic compounds)

  • 한 응엔 판 기우;김수희;김금진;최혁재;남두현
    • 미생물학회지
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    • 제54권4호
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    • pp.448-452
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    • 2018
  • 제주도 성산리 앞 바다 속 해면체로부터 분리한 Bacillus safensis KCTC 12796BP의 유전체를 분석하였다. 그 결과 3,935,874 bp의 환형 염색체와 36,690 bp의 plasmid 염기 서열을 확인하였다. 염색체는 G + C 함량이 41.4%로 75개의 위유 전자를 포함한 3,980개의 코딩 서열을, plasmid는 G + C 함량이 37.3%로 36개의 코딩 서열을 포함하고 있었다. 염색체 코딩 서열 중에는 81개의 tRNA 유전자, 24개 rRNA 유전자와 1개의 tmRNA 유전자가 있었다. 또한 포자 생성에 필요한 30개의 유전자, 포자피를 지령하는 16개의 유전자, 그리고 발아에 필요한 20개의 유전자도 발견되었다. 이외에 협막 다당체 생합성에 필요한 유전자와 편모 생합성 및 주화성에 필요한 유전자, 그리고 염 내성에 필요한 glycine-choline betaine 수송체에 관한 유전자도 존재하였다. 무엇보다도 항알러지활성을 보이는 이차대사산물 seongsanamide의 생합성을 지령하는 비리보좀성 펩타이드 합성효소 유전자를 확인할 수 있었다.

A Case Study of Shanghai Tang: How to Build a Chinese Luxury Brand

  • Heine, Klaus;Phan, Michel
    • Asia Marketing Journal
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    • 제15권1호
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    • pp.1-22
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    • 2013
  • This case focuses on Shanghai Tang, the first truly Chinese luxury brand that appeals to both Westerners and, more recently, to Chinese consumers worldwide. A visionary and wealthy businessman Sir David Tang created this company from scratch in 1994 in Hong Kong. Its story, spanned over almost two decades, has been fascinating. It went from what best a Chinese brand could be in the eyes of Westerners who love the Chinese culture, to a nearly-bankrupted company in 1998, before being acquired by Richemont, the second largest luxury group in the world. Since then, its turnaround has been spectacular with a growing appeal among Chinese luxury consumers who represent the core segment of the luxury industry today. The main objective of this case study is to formally examine how Shanghai Tang overcame its downfall and re-emerged as one the very few well- known Chinese luxury brands. More specifically, this case highlights the ways with which Shanghai Tang made a transitional change from a brand for Westerners who love the Chinese culture, to a brand for both, Westerners who love the Chinese culture and Chinese who love luxury. A close examination reveals that Shanghai Tang has followed the brand identity concept that consists of two major components: functional and emotional. The functional component for developing a luxury brand concerns all product characteristics that will make a product 'luxurious' in the eyes of the consumer, such as premium quality of cachemire from Mongolia, Chinese silk, lacquer, finest leather, porcelain, and jade in the case of Shanghai Tang. The emotional component consists of non-functional symbolic meanings of a brand. The symbolic meaning marks the major difference between a premium and a luxury brand. In the case of Shanghai Tang, its symbolic meaning refers to the Chinese culture and the brand aims to represent the best of Chinese traditions and establish itself as "the ambassador of modern Chinese style". It touches the Chinese heritage and emotions. Shanghai Tang has reinvented the modern Chinese chic by drawing back to the stylish decadence of Shanghai in the 1930s, which was then called the "Paris of the East", and this is where the brand finds inspiration to create its own myth. Once the functional and emotional components assured, Shanghai Tang has gone through a four-stage development to become the first global Chinese luxury brand: introduction, deepening, expansion, and revitalization. Introduction: David Tang discovered a market gap and had a vision to launch the first Chinese luxury brand to the world. The key success drivers for the introduction and management of a Chinese luxury brand are a solid brand identity and, above all, a creative mind, an inspired person. This was David Tang then, and this is now Raphael Le Masne de Chermont, the current Executive Chairman. Shanghai Tang combines Chinese and Western elements, which it finds to be the most sustainable platform for drawing consumers. Deepening: A major objective of the next phase is to become recognized as a luxury brand and a fashion or design authority. For this purpose, Shanghai Tang has cooperated with other well-regarded luxury and lifestyle brands such as Puma and Swarovski. It also expanded its product lines from high-end custom-made garments to music CDs and restaurant. Expansion: After the opening of his first store in Hong Kong in 1994, David Tang went on to open his second store in New York City three years later. However this New York retail operation was a financial disaster. Barely nineteen months after the opening, the store was shut down and quietly relocated to a cheaper location of Madison Avenue. Despite this failure, Shanghai Tang products found numerous followers especially among Western tourists and became "souvenir-like" must-haves. However, despite its strong brand DNA, the brand did not generate enough repeated sales and over the years the company cumulated heavy debts and became unprofitable. Revitalizing: After its purchase by Richemont in 1998, Le Masne de Chermont was appointed to lead the company, reposition the brand and undertake some major strategic changes such as revising the "Shanghai Tang" designs to appeal not only to Westerners but also to Chinese consumers, and to open new stores around the world. Since then, Shanghai Tang has become synonymous to a modern Chinese luxury lifestyle brand.

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미량화학(Small-Scale Chemistry)에 대한 초등학교 교사들의 인식 (A Survey of Perceptions of Elementary School Teachers on the Small-Scale Chemistry)

  • 김성규;공영태
    • 과학교육연구지
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    • 제34권2호
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    • pp.291-305
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    • 2010
  • 이 연구의 목적은 초등 교사들의 미량화학(Small-Scale Chemistry lab programs, SSC) 프로그램에 대한 교사의 인식을 알아보았다. SSC 연수 후 인식조사 참여 교사는 266명이다. 설문 내용은 SSC lab programs 교육과 적용의 필요성, 장점, 위험 및 안전, 환경오염과 처리, 경제성, 탐구능력 신장 등이다. 교사가 생각하는 전통적인 실험방법의 문제는 실험 준비시간이 많이 들고, 위험과 안전 그리고 사고에 대한 교사의 법적 책임, 경제적인 문제, 실험 후 폐기물 처리 어렵다, 자료 부족 순으로 답하였다. SSC를 경험한 교사는 거의 없었으며 SSC 연수 후 대부분의 교사들은 SSC 도입과 교육의 필요성에 긍정적으로 답하였다. 그리고 SSC 교육은 교과서 반영, 교사연수, 방과 후 교육, 강사 초청 순으로 하는 것이 좋겠다고 답하였다. SSC 교육에 필요자료로서는 CD, 비디오, 책자, 인쇄물 순으로 답하였다. SSC 장점을 묻는 설문에서는 단순, 편리, 시간절약, 다양성, 정성, 정량, 수업병행, 장난감 등 교사들 대부분이 긍정적으로 답하였다. 시약 등의 소량 사용으로 말미암아 폭발, 화재에 안전하다고 답하였고 플라스틱 사용으로 깨질 염려가 없어 안전하다고 답하였다. 또한 실험 후 처리가 쉽고 환경오염을 줄일 수 있겠다고 대부분이 긍정적으로 답하였다. 생활속 도구 사용으로 친숙하고 또한 장비구입이 용이하고 경제적이었다. 학습내용이해는 대체적으로 긍정적으로 답하였다. 개별 또는 2인 1조 편성으로 서로 충분한 토의가 부족하다고 생각하는 시각도 있었다. 문제해결력과 기술향상은 긍정적으로 답하였다. 특히, 창의력 신장에서 긍정적으로 답한 교사는 51.5% 였다. 대체적으로 탐구 능력 신장에는 교사들이 아직 체계화되지 않아 판단하는데 혼란스러운 것 같았다. 이 미량화학 프로그램이 초등현장에 간편하게 활용할 수 있게 교수-학습 자료의 개발을 위한 노력 및 보급 그리고 계속적인 연수가 이루어져서 초등 현장에 넓게 확대되었으면 한다.

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