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Cloning of a Gene Specifically Expressed During Early Stage of Fruiting Body Formation in Flammulina velutipes (팽이버섯의 자실체형성 초기과정에서 특이적으로 발현하는 유전자의 클로닝)

  • Kim, Dool-Yi;Azuma, Tomo-Nori
    • The Korean Journal of Mycology
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    • v.27 no.3 s.90
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    • pp.187-190
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    • 1999
  • A cDNA library was constructed using mRNA from the cells of 7-day-old cultures of Flammulina velutipes after induction of fruiting treatment. A cDNA clone, FVFD16 (Flammulina velutipes fruiting body differentiation), was selected by differential screening. The expression property of the FVFD16 gene was examined by Northern blot analysis. FVFD16 represents mRNA that is specifically expressed during differentiation of fruit bodies. The conspicuous accumulation of the FVFD16 mRNA was detected in 4-day-old and 1-day-old cultures. The nucleotide sequence of the FVFD16 gene was determined and the mRNA contained an open reading frame that encoded a putative protein of 128 amino acid residues (13.5 kDa).

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Analysis of Expressed Sequence Tags Generated from the Posterior Silkgland cDNA Clones of Antheraea yamamai (천잠 후부 견사선 유래 발현 유전자 꼬리표 작성 및 분석)

  • 윤은영;구태원;강석우;이혜원;황재삼;김호락
    • Journal of Life Science
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    • v.10 no.2
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    • pp.188-195
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    • 2000
  • In order to understand molecular events during silk synthesis and provide genetic resources for molecular breeding, we had analyzed the cDNA library constructed from the posterior silkgland of Antheraea yamamai and partially sequenced 276 randomly selected genes from the cDNA library. Database comparisons of the expressed sequence tags (ESTs) revealed that 26 non-redundant clones showed a high similarity with previously identified genes. Among them, 17 clones exhibited a homology with previously identified insect genes and 9 clones were identical to genes that were previously identified from other organisms. A functional categorization showed that silk synthesis-defense- or stress-related genes, as well as genes involved in the metabolic pathways and in the transcriptional or translational apparatus are represented. In this report, the clone (AY479) which had high similarity with fibroin from A. pernyi was particularly analyzed in detail. The AY479 clone was carboxyl terminal region of fibroin. The 472 bp cDNA has 123 amino acids that shared 85% homology with the fibroin from A. pernyi and its deduced peptide had unique feature, that is, sites of alanine rich residues.

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The Technical Services of the National Central Library: A Search for Rational Direction (국립중앙도서관의 자료정리현황과 그 방향에 관한 연구)

  • Lee Choon Hee
    • Journal of the Korean Society for Library and Information Science
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    • v.7
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    • pp.3-67
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    • 1980
  • Because of the changes made in the modes of cataloguing and classification in its long history, the present catalogue of the National Central Library has become complicated and provides an inadequate guide to its collection. There can be no doubt that this is a serious deficiency in a closed access library since materials housed in the library are virtually inaccessible to unskilled readers. The whole breakdown of the efficiency of the catalogue is emminent and will ultimately create the most serious problems for the library. The main purpose of this survey is: (a) to identify problem areas created by the frequent changes in the cataloguing and classifying principles in the library and (b) to grope a rational direction for the future development. Analysing the various classification schemes and cataloguing rules adopted in processing materials (mainly books) in the library, the following conclusions have been made. A. The library adopted five different clasification schemes in different periods, of which KDCP was used for the most part of its collection. KDCP is recommended to use for the future colletion. A classification development office is recommended to be established within the library, of which the main function is to revise the KDCP in collaboration with the appropriate committee of the Korean Library Association. B. The present practice in the library is to apply three different cataloguing rules and two different author notation tables to the Oriental, classical, and Western collections. Efforts should be made to find out an efficient system so that this variety is simplified. An alphabetical index should be added to the classified catalogue, and improvements are required in the Japanese collection. C. The technical services division is inadequately staffed. The staff should be sufficiently numerous and specially qualified. D. The present financial support for the technical services of the library is inadequate. Sufficient financial provision should be made to ensure the effective work. E. A feasibility study should be carried out to develop a computer processing system for providing machine-readable catalogue records on magnetic tape for use by the library community in Korea.

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Expressed Sequence Tags of Expression Profiles of Olive Flounder (Paralichthys olivaceus) Testis (ESTs (Expressed Sequence Tags)를 통한 넙치(Paralichthys olivaceus) 정소의 유전자 발현 패턴 분석)

  • Lee, Jeong-Ho;Kim, Jong-Hyun;Noh, Jae Koo;Kim, Hyun Chul;Kim, Young-Ok;Kim, Woo-Jin;Kim, Kyu-Won;Kim, Kyung-Kil
    • Korean Journal of Ichthyology
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    • v.19 no.4
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    • pp.257-265
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    • 2007
  • We constructed a cDNA library of testis from olive flounder (Paralichthys olivaceus) and a total of 248 expressed sequence tag (EST) clones were generated. In order to understand the molecular compositions of the olive flounder testis organs, the expression profiles of the identified clones in the cDNA library were analyzed. Gene annotation procedures and homology searches of the sequenced ESTs were locally done by BLASTX for amino acid similarity comparisons. Of the 248 EST clones, 156 ESTs showed significant homology to previously described genes while 92 ESTs were unidentified or novel. Comparative analysis of the 156 identified ESTs showed that 6 (3.8%) clones were representing 5 unique genes identified as homologous to the previously reported olive flounder ESTs, 100 (64.1%) clones representing 94 unique genes were identified as orthologs of known genes from other organisms, and orthologs were established for 50 (32.1%) clones representing 44 genes of known sequences with unknown functions. Furthermore, the testis library showed a more even distribution of cDNA clones with relatively fewer abundant clones that tend to contribute redundant clones in EST projects; thus, the testis library can supply more unique and novel cDNA sequences in olive flounder EST project.

Toxic dinoflagellate Gymnodinium catenatum Graham(Dinophyceae) from the southern coast of Korea: morphology, phylogeny and effects of temperature and salinity on growth (남해안에서 분리한 유독 와편모조류 Gymnodinium catenatum Graham (Dinophyceae): 형태, 분자계통학적 특성 및 온도와 염분에 따른 성장 특성)

  • Han, Kyong Ha;Li, Zhun;Kang, Byeong Jun;Youn, Joo Yeon;Shin, Hyeon Ho
    • Korean Journal of Environmental Biology
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    • v.37 no.1
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    • pp.31-41
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    • 2019
  • The toxic dinoflagellate Gymnodinium catenatum isolated from the southern coast of Korea was described under light and scanning electron microscopy, and its large subunit (LSU) rDNA was sequenced. In addition, the effects of temperature and salinity on its growth were investigated. The cells of G. catenatum, as viewed under the electronic microscope, were green-brown color, $38.1-77.4{\mu}m$ in length and $26.1-40.8{\mu}m$ in width. The epicone was conical, while the hypocone was trapezoidal. The nucleus was located at the central part of the cell. The apical groove was horseshoe-shaped and small pores were irregularly distributed on the cell surface. Molecular phylogeny based on LSU rDNA gene sequences showed that the Korean G. catenatum and previously reported species formed a monophyletic clade within Gymnodinium sensu stricto clade. The maximum growth rate of $0.37day^{-1}$, was obtained at $25^{\circ}C$ and 35 psu, and the maximum cell density of $1,073cells\;mL^{-1}$, was observed at $20^{\circ}C$ and 25 psu. However, G. catenatum did not grow at temperature < $15^{\circ}C$ and < $30^{\circ}C$. These results suggest that environmental conditions of summer and autumn in the southern coast of Korea may be favorable for the growth of G. catenatum.

Assembly of a Functional cDNA for Human Liver Growth Hormone Receptor: Cloning of Assembled hGHR cDNA (Human Liver로부터 Cloning한 cDNA성장호르몬 수용체의 기능성 검토)

  • 장규태;지선병홍;손동수;서원진삼;고교적웅
    • Journal of Embryo Transfer
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    • v.13 no.2
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    • pp.159-172
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    • 1998
  • 사람 성장호르몬 수용체(hGHR) cDNA는 PCR방법에 의하여 fagment로서 보고되어진 바 있으나, liver cDNA로 부터 전장을 cloning한 보고는 없는 실정으로 본 연구에서는 기능을 가진 약 4.6kbp의 cDNA hGHR을 cloning 하는데 성공하였다. 먼저 cloning하기 위하여 human liver mRNA와 human breast cancer tissue로부터 회수한 mRNA를 RT-PCR방법에 의하여 human cDNA library와 cloning에 필요한 probe를 제작하였다. human library mRNA는 GT-PCR방법에 의하여 증폭하여 증폭되어진 산물은 λZAP Vector를 이용하여 cDNA library를 구축하였고,screeing을 위하여 임 보고 되어진 hGHR fragment native sequence를 기초로 N-terminal부분의 primer를 설계하여 950bp의 probe를 얻는데 성공하였다. 이 probe를 이용하여 준비된 human liver cDNA library로부터 2.5$\times$10 6개의 plaque로부터 6개의 positive clone을 획득하였고, 이들중 poly Asignal인 "AATAAA"를 포함하고 있는 가장 긴 약 3.8kbp의 clone을 sequencing한 결과 open reading frame을 포함하고 있었으나, 5'부분의 결손되어 있었다. 그리하여 이 부분은 human breast cancer tissue로 부터 회수한 mRNA를 RT-PCR에 의하여 증폭하였고, sequencing결과 이미 보고되어진 native hGHR와 비교한 결과 하나의 nucleotide가 silent mutation으로 판명되었다.한편 human liver cDNA library로부터 cloning한 3.8cp의 positive clone의 5'end의 결손된 부분에 silent mutation된 PCR 산물을 연결함으로써 native hGHR와 유사한 cDNA hGHR subcloning에 성공하였다. 이러한 cDNA hGHR의 clone이 function을 가지고 있는지를 검토하기 위하여 eukaryotic 발현 vector인 pCXN2에 의거 ligation한 후 chinese hamster ovary cell[CHO-KI]에 transfect를 실시하였다. Dexamethasone은 첨가하지 않고 hGH만의 존재하에서 이들 cell을 배양시키고 cell menbrane에서 발현 여부를 판정키 위하여 hGHR monocloual antibody를 사용하여 flow cytometery해석을 실시하는 한편 125I-hGH binding assay에 의하여 hGH binding activity를 측정하였다. 최종적으로 GH signal transduction의 target genedf으로 알려져 있는 serine protease inhibitor 2.1(Spi 2.1) gene의 promotor activity를 검토한 결과 hGHR을 transfect한 CHO Cell에 있어서 hGH의 농도에 의존적으로 증가되었다. 따라서 본 실험에서 cloning한 cDNA hGHR는 native hGHR와 같은 기능을 가지는 것으로 판명되었다.것으로 판명되었다.

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Identification of Non-Muscle Nebulin Isoform in Human Brain Library

  • Joo, Young-Mi;Lee, Min-A;Choi, Pyung-Rak;Choi, Jae-Kyoung;Lee, Yeong-Mi;Choi, Su-Il;Kim, Myong-Shin;Jeon, Eun-Hee;Kim, So-Young;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.10 no.1
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    • pp.23-29
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    • 2004
  • Nebulin is a (Mr 600∼900 kDa) large actin-binding protein specific to skeletal muscle and thought to act as a molecular template that regulates the length of thin filaments. Cardiac muscles of higher vertebrates have been shown earlier to lack nebulin. Recently, full-length nebulin mRNA transcripts have been detected in heart muscle, but at lower levels than in skeletal muscle. Nebulin expression also was detected in the kidney, eye, and otic canal, suggesting that nebulin isoforms may also be expressed in these organs. We have searched for nebulin isoforms in brain of human using PCR and Northern blot. Here, we provide evidence that nebulin mRNA transcripts are expressed in brain. Seven nebulin isoforms (B, C, D, E, F, G and H form) are obtained in human skeletal muscle and four isoforms (B, C, G and H form) in human brain cDNA library. We cloned the 1.3 kb of nebulin fragment from human adult brain library by PCR. The identity of the PCR product was confirmed by sequence analysis. The partial brain nebulin sequence was 99% identical to the skeletal muscle cDNA as determined by Blast alignment. It contains two simple-repeats HR1, HR2 and linker-repeats exon l35∼143 except exon 140. It was different from skeletal muscle B form, which contain HR1 and HR8. These data suggest that nebulin isoform diversity occurs even more extensively than previously known, likely contributing to the distinct thin filament architecture of different striated muscles.

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Let Us Make: Interrogating Personnel's Perception towards Makerspace in a Nigerian University Library

  • Igbinovia, Magnus Osahon;Alex-Nmecha, Juliet C.
    • International Journal of Knowledge Content Development & Technology
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    • v.11 no.2
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    • pp.23-36
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    • 2021
  • Makerspace offers an innovative way for university libraries to render services and achieve its core objectives. However, there is low level of adoption in Nigerian university libraries, hence the need to investigate the personnel's perception towards makerspace as an innovative learning platform. To achieve this, the descriptive research design was used to elicit data with the aid of a structured questionnaire, from the 96 library personnel in John Harris Library, University of Benin. Of the 96 copies of questionnaire distributed, 68 copies were returned and analyzed using descriptive statistics. The results showed that the library personnel were familiar with makerspace even though they still require more information about it. Also, that makerspace gives students the opportunity to acquire ew skill and enhances collaboration among learners. Moreover, the study revealed that budget constraints, high cost and maintenance of equipments, erratic power supply and lack of staff training are some of the perceived challenges to makerspace implementation. Consequently, the study revealed that university libraries should strategize on how to increase allocation of funds, organize makerspace webinar for personnel, ensure stable electricity supply and create a dedicated space for makerspace in the library. In conclusion, makerspace will provide students with the technological exposure and creative abilities required for the future, and as such effective planning should be made for its deployment and sustainability in Nigerian university libraries.

Cloning of Epidermis-specific cDNAS Encoding a Lipid Transfer Protein and an Aldehyde Decarbonylase from Senecio odorus

  • Pyee, Jaeho
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.189-195
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    • 1996
  • The major cuticular components have been shown to be synthesized in the epidermis. Therefore, cloning of epidermis-specific genes could yield information to be used to isolate and characterize the enzymes involved in the cuticle biosynthesis. A subtractive cDNA library was prepared from Senecio odorus in which epidermis-specific cDNAs were enriched. Differential screening of the library using epidermal and non-epidermal probes revealed two cDNAs. One of them designated epi425 was identified, based on the sequence homology, as a member of a new class in the LTP gene family and the other clone designated epi23 as a gene encoding an aldehyde decarbonylase. Northern blot analyses showed that epi425 and epi23 cDNAs hybridized with a transcript of about 600 and 2, 100 nucleotides, respectively, from the epidermis but not from the non-epidermal tissues. Further characterization of these clones will provide more information on the mechanism of the cuticle biosynthesis.

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Construction of Full-Lenth cDNA Library from Seosan 6-pieces Gallic and cDNA Cloning of Allinase and Lectin Genes (서산 6쪽마늘의 Full-lenth cDNA library 구축 및 allinase와 lectin 유전자의 cDNA 클로닝)

  • Lee, Mi-Ok;Kim, Hae-Kyung;Lee, Jin-Sung
    • Proceedings of the KAIS Fall Conference
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    • 2007.05a
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    • pp.270-272
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    • 2007
  • 본 연구는 서산 6쪽 마늘로부터 완전장 유전자 은행의 제작과 이를 통해서 확보된 1,000여개 재조합 클론에 대한 염기서열 결과를 web-based database를 통한 상동성 분석으로 부터 서산 마늘의 발현 유전자에 대한 생물정보학적 분석에 관해 것이며 본 연구로 부터 마늘의 대표적 생리활성 물질인 allicin의 생성에 관여하는 효소인 allinase의 cDNA를 클로닝 및 완전 염기서열을 해석하였으며 allinase 유전자의 genomic structure 에 대한 일부의 결과를 확보하였다. 또한 다양한 생물종에서 연구 되어지고 있는 생리활성 단백질인 lectin 유전자 cDNA를 클로닝하여 완전 염기서멸을 분석하고, 6xHis Tag올 통한 재조합 단백질을 대장균에서 E.coli에서 발현시켰다.

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