• Title/Summary/Keyword: C Library

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Selection and identification of single-domain antibody against Peste des Petits Ruminants virus

  • Liu, Dan;Li, Lingxia;Cao, Xiaoan;Wu, Jinyan;Du, Guoyu;Shang, Youjun
    • Journal of Veterinary Science
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    • v.22 no.4
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    • pp.45.1-45.13
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    • 2021
  • Background: Peste des petits ruminants (PPR) is an infectious disease caused by the peste des petits ruminants virus (PPRV) that mainly produces respiratory symptoms in affected animals, resulting in great losses in the world's agriculture industry every year. Single-domain variable heavy chain (VHH) antibody fragments, also referred to as nanobodies, have high expression yields and other advantages including ease of purification and high solubility. Objectives: The purpose of this study is to obtain a single-domain antibody with good reactivity and high specificity against PPRV. Methods: A VHH cDNA library was established by immunizing camels with PPRV vaccine, and the capacity and diversity of the library were examined. Four PPRV VHHs were selected, and the biological activity and antigen-binding capacity of the four VHHs were identified by western blot, indirect immunofluorescence, and enzyme-linked immunosorbent assay (ELISA) analyses. ELISA was used to identify whether the four VHHs were specific for PPRV, and VHH neutralization tests were carried out. ELISA and western blot analyses were used to identify which PPRV protein was targeted by VHH2. Results: The PPRV cDNA library was constructed successfully. The library capacity was greater than 2.0 × 106 cfu/mL, and the inserted fragment size was approximately 400 bp to 2000 bp. The average length of the cDNA library fragment was about 1000 bp, and the recombination rate was approximately 100%. Four single-domain antibody sequences were selected, and proteins expressed in the supernatant were obtained. The four VHHs were shown to have biological activity, close affinity to PPRV, and no cross-reaction with common sheep diseases. All four VHHs had neutralization activity, and VHH2 was specific to the PPRV M protein. Conclusions: The results of this preliminary research of PPRV VHHs showed that four screened VHH antibodies could be useful in future applications. This study provided new materials for inclusion in PPRV research.

Optimized Serological Isolation of Lung-Cancer-associated Antigens from a Yeast Surface-expressed cDNA Library

  • Kim, Min-Soo;Choi, Hye-Young;Choi, Yong-Soo;Kim, Jhin-Gook;Kim, Yong-Sung
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.993-1001
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    • 2007
  • The technique of serological analysis of antigens by recombinant cDNA expression library (SEREX) uses autologous patient sera as a screening probe to isolate tumor-associated antigens for various tumor types. Isolation of tumor-associated antigens that are specifically reactive with patient sera, but not with normal sera, is important to avoid false-positive and autoimmunogenic antigens for the cancer immunotherapy. Here, we describe a selection methodology to isolate patient sera-specific antigens from a yeast surface-expressed cDNA library constructed from 15 patient lung tissues with non-small cell lung cancer (NSCLC). Several rounds of positive selection using patient sera alone as a screening probe isolated clones exhibiting comparable reactivity with both patient and normal sera. However, the combination of negative selection with allogeneic normal sera to remove antigens reactive with normal sera and subsequent positive selection with patient sera efficiently enriched patient sera-specific antigens. Using the selection methodology described here, we isolated 3 known and 5 unknown proteins, which have not been isolated previously, but and potentially associated with NSCLC.

A Study on User Satisfaction Research about Spacial Composition before and after Remodeling University Library Focused on C University Library (대학도서관 리모델링 전.후의 공간구성에 관한 이용자 만족도 조사 연구 - C대학교 중앙도서관을 중심으로 -)

  • Nam, Young-Joon;Moon, Jung-Hyun;Yi, Hyun-Jung
    • Journal of the Korean BIBLIA Society for library and Information Science
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    • v.20 no.4
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    • pp.205-222
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    • 2009
  • University libraries are involved in college education and college education is also involved in university libraries. To the knowledge information society increases the competitive power of the university and the academic library which national competitive power strengthens further must be operated developmentally. Professors and students to support research and learning activities appropriate facilities and systems must be equipped with features and the role of university libraries will be fulfilled properly. In this study, we analyze changes in the configuration space, and accordingly remodel before and after the user satisfaction survey of spatial configuration on the users' satisfaction and the results are to analyze based on C University Library. As a result of this study, the library space was utilized to determine the importance of purpose in the future, these results increase the quality of library service or library user satisfaction survey. Study will be a good reference for researchers.

A Study on Information Literacy Education Program of Medical Library: Focused on C University Medical Library (의학도서관 정보이용교육 프로그램 연구: C대학교 의학도서관을 중심으로)

  • Lee, Yong-Jae;Woo, Sang-Hee
    • Journal of Information Management
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    • v.39 no.3
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    • pp.49-71
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    • 2008
  • This study aims to develop more user-oriented information literacy education for medical libraries in Korea. For this purpose, we benchmarked some advanced models in and out of Korea, and examined a case of C medical library which is medium size in Korea. Especially, we looked into the actual condition of information literacy education of C medical library and the information need of its users. We suggested some ideas to construct the programs for information literacy education of medical libraries in Korea. Especially, we proposed some considerations, contents and flow for the programs. The results of this study can be useful references whenever each medical library in Korea will start or make up for its information literacy program.

Apoptosis Suppressor에 관련된 유전자 스크린 방법과 동정된 유전자 특성 규명

  • 황규찬;옥도원;권득남;신혜경;김진회
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.16-16
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    • 2001
  • Apoptosis로 일컬어지는 예정된 세포사멸(programmed cell death)은 개별 세포의 입장에서는 곧바로 사멸을 의미하지만, 정상적인 고등 생물의 입장에서는 개체의 발생과 분화하는데 프로그램된 과정이다. 자발적 세포사멸은 다른 조직에 비해 생식 조직인 난소나 정소에서 복잡한 apoptosis 기작들을 가지리라 사료된다. 본 연구는 Bcl-2 family중 apoptotic protein인 Bax에 대해 suppression하는 유전자를 yeast system을 활용하여 돼지 정소와 난소로부터 각각 cDNA library를 구축한 후 탐색하였다. 탐색에 활용된 cDNA library는 돼지의 정소와 난소로부터 mRNA를 분리하여 yeast vector인 pAD-GAL4-2.1에 구축하였고, 마우스 bax 유전자는 gal 1 promoter의 조절 하에 glucose 배지에서는 유도되지 않고, galactose 배지에서만 선택적으로 Bax를 발현할 수 있는 효모 vector(pL19-bax)를 구축하였다. Bax에 의한 apoptosis suppressor를 탐색하기 위해 우선 효모 W303에 pL19-bax를 transform하여 glucose 배지에서 Bax의 발현을 억제하였다. pL19-bax를 가진 효모에 정소와 난소로부터 구축된 cDNA library를 transform 시키고, transform된 효모는 각각 Bax에 의한 toxicity를 저해하는 유전자를 찾기 위해 스크린되었다. 이러한 방법으로 정소 cDNA library 탐색에서는 5 $\times$ $10^{6}$ transformant중 39개, 난소cDNA library 탐색에서는 2 $\times$ $10^{6}$ transformant중 26개의 콜로니가 생존하였다. 이들 콜로니로부터 유전자를 분리하여 분석해 본 결과 여러 그룹으로 분류할 수 있었다. 각 그룹의 관련 유전자는 protein synthesis/degradation 12종, oxidation/reductation 5종, detoxin/ cell cycle promoter 3종, signal transduction/growth factor 5종, 그리고 알려지지 않은 유전자 9종이었다. 그 중, bax-toxicity inhibition에 강력한 survival phenotype을 가지는 유전자(pSEDL)를 동정하였다. 이것은 T3-4-1 콜로니로부터 분리하였는데 140개 아미노산으로 이루어진 인간 SEDL(GenBank, XM_013096) 유전자와 매우 유사한 homology를 가지며, bax와 관련된 기능은 밝혀져 있지 않다. 이외에도 분리된 유전자에는 NADH, thioreduction, 그리고 cytochrome oxidase와 같은 positive 유전자 군이 크로닝되어, Bax를 이용한 효모에서 apoptosis suppressor에 관련된 유전자를 손쉽게 스크린하는 것이 가능하고, 분리된 유전자의 기능을 예측할 수 있어 지금까지 보고된 유전자 크로닝법 보다는 강력한 수단으로 활용될 수 있다는 사실을 시사하였다. 그러나, ORF에 관계없이 Bax 발현에 저항하는 유전자군이 선발된다든지 하는 문제점은 금후 검토가 필요하리라 사료된다.

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A Study on the Typology of Mixing Activities and Space Organization in Public Library Buildings (공공도서관의 공간구성과 복합화 유형에 관한 연구)

  • 김정희;이상호
    • Korean Institute of Interior Design Journal
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    • no.41
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    • pp.146-154
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    • 2003
  • The purpose of this study is, According to society changing fast, human life is changing in the direction of complexity and variety, the middle class and multiple structure have been generated in relation between architecture and function. Accordingly, considering the analyzed elements of the main floor space planning and relation of the factors in space planning of Seoul s public libraries. The Public Library is variety of function is abstracted to the plan for Space Composition Form from future public library, analysis of “This point of user, how to composition is the space of different function” and “How to is the connection between the educational function and the other function, complexity of space.” With above distinctive themes and the research, the typology of public library in seoul city, At the first, the research shows that the public library is 8 type, and high in typology of “TYPE 7 (A3-B2-Cl)”. Secondary, There appears public library, such as in the satisfaction of the One-Function-User is high in “TYPE 3 (A1-B2-C3)”, in the satisfaction over the Two-Function-User is high in “TYPE 6 (A3-B1-C1)”

A Study on the Molecular Biology of Renin-Angiotensin System : Renin Gene Expression and Construction of Genomic Library (Renin-Angiotensin계의 분자생물학적 연구 : Renin유전자의 발현과 Genomic Library작성)

  • 박영순;한동민;김종호;문영희;이호섭;고건일;김성준
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.35-44
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    • 1990
  • Poly (A) + RNA was isolated from mouse submaxillary gland and renin mRNA was isolated by poly (U)-sepharose chromatography and sucrose linear densiW gradient centifugation. And renin mRNA was identified by in vitro translation and immunoprecipitation. In order to construct recombinant plasmid, renin cDNA was synthesized by using reverse transcriptase and inserted into EcoRi site of PUC19. In addition, the cDNA was also synthesized using polymerase chain reaction and inserted into HindlIl site of PUC19. The recombinant plasmid was transformed into JMlO3 and the expression of the inserted renin cDNA was examined. The transformant produced renin protein having a molecular weight of 45, 000 dolton, which showed hypertensive effect upon injecting it into rabbit ear vein. A renin genomic library was prepared by inserting rabbit kidney DNA into EMBL3 phage, and was screeined for the isolation of renin gemomic DNA using renin cDNA probe.

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Unusual Orientation of cDNAs Found in a cDNA Library

  • Lee Jeongyeo;Song Hayoung;Lim Yong-Pyo;Hur Yoonkang
    • Journal of Plant Biotechnology
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    • v.7 no.1
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    • pp.51-55
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    • 2005
  • Many cloning vectors in which cDNAs can be inserted to the sense orientation have been developed. Uni-ZAP XR vector (Stratagene) should contain clones that are oriented to sense direction with respect to T3 RNA polymerase primer. Unexpectedly, large portions of cDNAs in Chinese cabbage cDNA library showed unusual insertions, antisense orientation and a hybrid of two different clones. Using two clones, 4H03 and 53-B10, derived from different cDNA libraries, we proposed and demonstrated the possibility of unusual-construct formation by in vitro translation and northern blot analysis. The 4H03 clone was inserted with inverse direction, and its transcript and translation product could be produced by T7 RNA polymerase, indicating that this clone is definitely inserted into inverse orientation. The 53-B10 that contains two independent genes was turned out to be a hybrid in which two genes are inserted to opposite direction each other. All unusual constructs might be due to the presence of small fragments of DNA, like adapter. However, the mechanism underlined the formation of unusual constructs is still remain to be solved.

Gene Analysis Related Energy Metabolism of Leaf Expressed Sequence Tags Database of Korean Ginseng (Panax ginseng C.A. Meyer) (고려인삼(Panax ginseng C.A, Meyer)의 잎 ESTs database에서 Energy 대사 관련 유전자 분석)

  • Lee Jong-Il;Yoon Jae-Ho;Song Won-Seob;Lee Bum-Soo;In Jun-Gyo;Kim Eun-Jeong;Yang Deok-Chun
    • Korean Journal of Plant Resources
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    • v.19 no.1
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    • pp.174-179
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    • 2006
  • A cDNA library was constructed from leaf samples of 4-year-old Panax ginseng cultured in a field. 3,000 EST from a size selected leaf cDNA library were analyzed. The 349 of 2,896 cDNA clones has related with energy metabolism genes. The 349 known genes were categorized into nine groups according to their functional classification, aerobic respiration(48.4%), accessory proteins of electron transport and membrane associated energy conservation(17.2%), glycolysis and gluconeogenesis(3.4%), electron transport and membrane associated energy conservation(2.9%), respiration(2.0%), glycolysis methylglyoxal bypass(1.7%), metabolism of energy reserves(0.6%) and alcohol fermentation(0.3%).