• Title/Summary/Keyword: cell library

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Morphological Features of Marine Dinoflagellates from Jangmok Harbour in Jinhae Bay, Korea: A Case of 30 Species in the Orders Prorocentrales, Dinophysiales, Gonyaulacales and Gymnodiniales

  • Shin, Hyeon Ho;Kim, Eun Song;Li, Zhun;Youn, Joo Yeon;Jeon, Seul Gi;Oh, Seok Jin
    • Korean Journal of Environmental Biology
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    • v.34 no.3
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    • pp.141-150
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    • 2016
  • Most previous studies on dinoflagellates in Korean coastal areas were conducted without morphological descriptions and illustrations of the observed dinoflagellates. This indicates that the species and diversity of dinoflagellates may have been respectively misidentified and underestimated in the past, probably due to cell shrinkage, distortion and loss caused by sample fixation. This study provides information on the morphological observations of four dinoflagellate orders (Prorocentrales, Dinophysiales, Gonyaulacales and Gymnodiniales) from Jangmok Harbour in Jinhae Bay, Korea. The unfixed samples were collected weekly from December 2013 to February 2015. A total of 13 genera and 30 species were identified using light and scanning electron microscopy, although some samples were not clarified at the species level. Harmful dinoflagellates, Prorocentrum donghaiense, Tripos furca, Alexandrium affine, A. fundyense, Akashiwo sanguinea and Cochlodinium polykrikoides, were identified based on the morphological observations. The results also reflect the occurrence and identification of dinoflagellates that had not been previously recorded in Jangmok Harbour.

Role of CAGE, a Novel Cancer/Testis Antigen, in Various Cellular Processes, Including Tumorigenesis, Cytolytic T Lymphocyte Induction, and Cell Motility

  • Kim, Young-Mi;Jeoung, Doo-Il
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.600-610
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    • 2008
  • A cancer-associated antigen gene (CAGE) was identified by serological analysis of a recombinant cDNA expression library (SEREX). The gene was identified by screening cDNA expression libraries of human testis and gastric cancer cell lines with sera from patients with gastric cancer. CAGE was found to contain a D-E-A-D box domain and encodes a putative protein of 630 amino acids with possible helicase activity. The CAGE gene is widely expressed in various cancer tissues and cancer cell lines. Demethylation plays a role in the activation of CAGE in certain cancer cell lines where the gene is not expressed. The functional roles of CAGE in tumorigenesis, the molecular mechanisms of CAGE expression, and cell motility are also discussed.

Expressed Sequence Tag Analysis of Toxic Alexandrium tamarense and Identification of Saxitoxin Biosynthetic Genes (독성 Alexandrium tamarense 의 EST 분석 및 삭시톡신 생합성 유전자의 확인)

  • Chang, Man;Lee, Juyun;Chung, Youngjae;Lee, Gunsup;Kim, Dongguin;Lee, Taek-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.14 no.7
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    • pp.3582-3588
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    • 2013
  • Expressed sequence tag (EST) library was constructed from A. tamarense. Base sequences of EST clones were analyzed and saxitoxin biosynthesis-related genes were cloned. Sequences of 827 clones were analyzed and 564 EST were functionally clustered using Blast searches against GenBank. Main genes in the EST had functions on cellular organization, cell metabolism, energy, cell cycle and DNA processing, cellular transport and transport, cell rescue, defense, death and aging, and transcription. Moreover, expression of S-adenosylmethionine synthetase and H2A histone family genes were increased in the toxic A. tamarense. These results show that two genes could be a good biomarkers for the detection of saxitoxin biosynthesis in the A. tamarense.

Isolation and Characterization of $A{\alpha}$ mating locus from Schizophyllum commune (치마버섯(Schizophyllum commune)으로부터 $A{\alpha}$ mating locus의 분리 및 특성)

  • Park, Dong-Chul;Novotny, Charles P.;Ullich, Robert C.;Lee, Kap-Duk;Lee, Kap-Rang
    • The Korean Journal of Mycology
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    • v.22 no.3
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    • pp.247-253
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    • 1994
  • This study was carried out to isolate and characterize $A{\alpha}$ mating locus controlling fruiting body formation directly in the Basidiomycete Schzophyllum commune growing in the North America. Total numbers of genomic library of S. commune UVM1-34 was about $2{\times}10^4$ cells. About 90% library was appeared to have about 35 kb inserted genome DNA in cosmid pTC20 vector. 6 clones were proved to have positive signal to probes within Z and Y region in colony and southern hybridization. In the mating activity test, all the 6 positive clones were appeared to have $A{\alpha}3$ mating activity although they had two different restriction patterns. pSC13 containing 5.7 Kb PstI-fragment of UVM 1-34 $A{\alpha}3$ allele showed about 50% clamp cell formation indicating mating activity when cotransformation was done together with cosmid pTC20.

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Definition of the peptide mimotope of cellular receptor for hepatitis C virus E2 protein using random peptide library (Random peptide library를 이용한 C형 간염바이러스 E2 단백질 세포막 수용체의 peptide mimotope 규명)

  • Lee, In-Hee;Paik, Jae-Eun;Seol, Sang-Yong;Seog, Dae-Hyun;Park, Sae-Gwang;Choi, In-Hak
    • IMMUNE NETWORK
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    • v.1 no.1
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    • pp.77-86
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    • 2001
  • Background: Hepatitis C virus(HCV), a family of Flaviviridae, has a host cell-derived envelope containing a positive-stranded RNA genome, and has been known as the maj or etiological agent for chronic hepatitis, hepatic cirrhosis, and hepatocellular carcinoma. There remains a need to dissect a molecular mechanism of pathogenesis for the development of therapeutic and effective preventive measure for HCV. Identification of cellular receptor is of central importance not only to understand the viral pathogenesis, but also to exploit strategies for prevention of HCV. This study was aimed at identifying peptide mimotopes inhibiting the binding of E2 protein of HCV to MOLT-4 cell. Methods: In this study, phage peptide library displaying a random peptides consisting of 7 or 12 random peptides was employed in order to pan against E2 protein. Free HCV particles were separated from the immune complex forms by immunoprecipitation using anti-human IgG antibody, and used for HCV-capture ELISA. To identify the peptides inhibiting E2-binding to MOLT-4 cells, E2 protein was subj ect to bind to MOLT-4 cells under the competition with phage peptides. Results: Several phage peptides were selected for their specific binding to E2 protein, which showed the conserved sequence of SHFWRAP from 3 different peptide sequences. They were also able to recognize the HCV particles in the sera of HCV patients captured by monoclonal antibody against E2 protein. Two of them, showing peptide sequence of HLGPWMSHWFQR and WAPPLERSSLFY respectively, were revealed to inhibit the binding of E2 protein to MOLT-4 cell efficiently in dose dependent mode. However, few membrane-associated receptor candidates were seen using Fasta3 programe for homology search with these peptides. Conclusion: Phage peptides containing HLGPWMSHWFQR and WAPPLERSSLFY respectively, showed the inhibition of E2-binding to MOLT-4 cells. However, they did not reveal any homologues to cellular receptors from GenBank database. In further study, cellular receptor could be identified through the screening of cDNA library from MOLT-4 or hepatocytes using antibodies against these peptide mimotopes.

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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.

Integration of Single-Cell RNA-Seq Datasets: A Review of Computational Methods

  • Yeonjae Ryu;Geun Hee Han;Eunsoo Jung;Daehee Hwang
    • Molecules and Cells
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    • v.46 no.2
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    • pp.106-119
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    • 2023
  • With the increased number of single-cell RNA sequencing (scRNA-seq) datasets in public repositories, integrative analysis of multiple scRNA-seq datasets has become commonplace. Batch effects among different datasets are inevitable because of differences in cell isolation and handling protocols, library preparation technology, and sequencing platforms. To remove these batch effects for effective integration of multiple scRNA-seq datasets, a number of methodologies have been developed based on diverse concepts and approaches. These methods have proven useful for examining whether cellular features, such as cell subpopulations and marker genes, identified from a certain dataset, are consistently present, or whether their condition-dependent variations, such as increases in cell subpopulations in particular disease-related conditions, are consistently observed in different datasets generated under similar or distinct conditions. In this review, we summarize the concepts and approaches of the integration methods and their pros and cons as has been reported in previous literature.

Expression of Human Serine Palmitoyltransferase Genes for Antibody Development (Antibody 제작을 위한 human serine palmitoyltransferase 유전자의 발현)

  • 김희숙
    • Journal of Life Science
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    • v.14 no.2
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    • pp.315-319
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    • 2004
  • For antibody development of human serine palmitoyltransferase (SPT, EC 2.3.1.50), SPTLC1 and SPTLC2 genes were subcloned in pRset vector and expressed in E. coli BL21 (DE3)pLys cells. Eucaryotic SPT is a membrane-bound heterodimer enzyme, while all other members are soluble homodimer enzymes. cDNA library were obtained from total RNA from human embryo kidney cell line, HEK293, using RT-PCR and PCR with specific primers was carried out for preparing SPTLC1 and SPTLC2 genes. pRset vector which can express hexahistidine-tag fusion protein was used and the DNA sequences of pRsetB/SPTLC1 and pRsetA/SPTLC2 were confirmed. Recombinant BL21 cells with SPTLC subunits were selected with LB plate containing ampicillin and chroramphenicol. SPTLC1 and SPTLC2 proteins were induced with 1 mM IPTG and seperated on 10% SDS-PAGE gel. Expressed proteins were confirmed by western blotting with His-tag antibody.

Niclosamide induces mitochondria fragmentation and promotes both apoptotic and autophagic cell death

  • Park, So-Jung;Shin, Ji-Hyun;Kang, Hee;Hwang, Jung-Jin;Cho, Dong-Hyung
    • BMB Reports
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    • v.44 no.8
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    • pp.517-522
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    • 2011
  • Mitochondrial dynamics not only involves mitochondrial morphology but also mitochondrial biogenesis, mitochondrial distribution, and cell death. To identify specific regulators to mitochondria dynamics, we screened a chemical library and identified niclosamide as a potent inducer of mitochondria fission. Niclosamide promoted mitochondrial fragmentation but this was blocked by down-regulation of Drp1. Niclosamide treatment resulted in the disruption of mitochondria membrane potential and reduction of ATP levels. Moreover, niclosamide led to apoptotic cell death by caspase-3 activation. Interestingly, niclosamide also increased autophagic activity. Inhibition of autophagy suppressed niclosamide-induced cell death. Therefore, our findings suggest that niclosamide induces mitochondria fragmentation and may contribute to apoptotic and autophagic cell death.

In Vitro Selection of Cancer-Specific RNA Aptamers

  • Lee Young-Ju;Lee Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1149-1153
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    • 2006
  • In this study, nuclease-resistant RNA aptamers that are specific for Jurkat T leukemia cells were selected by a subtractive systemic evolution of ligands by exponential enrichment (SELEX) method. A randomized nuclease-resistant RNA library was incubated with normal peripheral blood mononuclear cells (PBMC) in each round to preclude RNAs that recognize the common cellular components on the surface of normal and cancer cells. The precluded RNAs were used for the selection of Jurkat T cell-specific aptamers, and the specific RNAs were then gradually enriched from start to the following selections. After 16 rounds of the subtractive SELEX, the selected aptamers were found to preferentially bind to Jurkat T cells, but not to the normal PBMC, evidenced by fluorescence-activated cell sorting analysis. Thus, the subtractive SELEX can be used to identify ligands to cancer-specific biological markers without prior knowledge of the nature of markers. The aptamers could be applied to specific cell sorting, tumor therapy, and diagnosis, and moreover, to find cancer cell-specific markers.