• Title/Summary/Keyword: molecular line

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Re-identification of Two Tonguefishes (Pleuronectiformes) from Korea using Morphological and Molecular Analyses (형태 및 분자분석에 의한 한국산 참서대과 어류(가자미목) 2종의 재동정)

  • Kwun, Hyuck Joon;Kim, Jin-Koo
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.49 no.2
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    • pp.208-213
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    • 2016
  • The re-identification of two Korean tonguefishes, Cynoglossus interruptus and Symphurus orientalis, was carried out using eight specimens collected from Korean waters in 2007 and 2013. C. interruptus is characterized by having a single row of scales between rows connected to the supraorbital line and the middle lateral line, 107–113 dorsal fin rays, 86–89 anal fin rays, and 53–55 vertebrae. S. orientalis is characterized by having a 1-2-2-2-2 ID pattern, 97–100 dorsal fin rays, 83–89 anal fin rays, and 52–55 vertebrae. Molecular analysis using mitochondrial DNA Cytochrome Oxidase subunit I sequences showed that specimens of the two species corresponded well to Japanese C. interruptus and Taiwanese S. orientalis, respectively. Therefore, although several reports have raised questions regarding the distribution of C. interruptus and S. orientalis in Korean waters, morphological and molecular data confirm that these two species are indeed distributed in these waters.

Endogenous Gene Expression of p53 and Regulatory Subunits of Cyclic AMP-dependent Protein Kinase in Ovarian Cancer Cells (난소암 세포주에서 p53과 Cyclic AMP-dependent Protein Kinase의 Regulatory Subunit 유전자들의 발현에 관한 연구)

  • Jin Seo;Park, Woonmee;Hwang, Eun-Seong;Lee, Je-Ho;Hong, Seung-Hwan
    • The Korean Journal of Zoology
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    • v.38 no.2
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    • pp.204-211
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    • 1995
  • In an effort to develop a new therapeutic strategy for human gene therapy of solid ovarian tumor, we studied the expression of the p53 tumor suppressor Sene as well as regulatory subunits of cyclic AMP (cAMP)-dependent protein kinase in human ovarian carcinoma cells. Four cell lines (2774, Caov-3, SK-OV-3 and OVCAR-3) were selected for the analyses. The p53 transcript and protein were detected only in the 2774 cell line by Northern and Western Bnalysis. In the relatively fast growing cell line, SK-OV-3, the %rope 1 a regulstorv subunit (RIA of CAMP-dependent protein kinase was the highest among the four cell lines. The expression level of $RII\beta$ protein was low in the four cell lines examined. These results maw point to a direction to select the target gene(sl to be employed for gene therapy to control the ovarian cancer.

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Live Cell Detection of Monoclonal Antibody Light and Heavy Chain mRNAs using Molecular Beacons (분자 비컨을 이용한 살아 있는 세포에서 단일클론항체 경쇄와 중쇄 mRNA 검출에 의한 세포주 선별방법)

  • Jeong, Seunga;Rhee, Won Jong
    • KSBB Journal
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    • v.31 no.1
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    • pp.33-39
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    • 2016
  • Developing the method for the selection of animal cell line producing therapeutic monoclonal antibody (mAb) is invaluable as its market is rapidly growing. Although the quality of produced mAb is as important as quantity, however there is no method developed for the selective screening of cell lines on the basis of both quantity and quality. From recent reports, the ratio of light and heavy chain mRNAs of mAb in the cell is a key parameter for the indication of product quality. Therefore, it is obvious that developing the novel method that can detect both light and heavy chain mRNAs in single live cell will provide unprecedented opportunities in bio-industry. Here, we have constructed oligonucleotide probes, molecular beacons for the detection of light or heavy chain mRNAs, respectively, in the live cells producing mAbs. Both beacons showed increased fluorescent intensity after transient transfection of plasmid expressing mAbs analyzed by fluorometer. Flow cytometric analysis clearly demonstrated that both molecular beacons can simultaneously detect the expression of light and heavy chain mRNAs of mAb in the same cell. The technique described in the thesis provides the new direction and concept for developing the method for the smart selection of cell lines producing recombinant proteins including therapeutic mAbs.

Characteristic Chemical Correlations in Nearby Star-forming Molecular Clouds

  • Yun, Hyeong-Sik;Lee, Jeong-Eun;Evans, Neal J. II;Offner, Stella;Heyer, Mark H.;Choi, Yunhee;Lee, Yong-Hee;Baek, Giseon;Choi, Minho;Kang, Hyunwoo;Tatematsu, Ken'ichi;Lee, Seokho;Yang, Yao-Lun;Gaches, Brandt;Chen, How-Huan
    • The Bulletin of The Korean Astronomical Society
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    • v.45 no.1
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    • pp.41.1-41.1
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    • 2020
  • Different molecular lines trace different physical environments (with various densities and temperatures) within molecular clouds (MCs). Therefore, multimolecular line observations are crucial to study the physical and chemical structures of MCs. We observed the Orion A and Ophiuchus clouds in six different molecular lines as a Taeduk Radio Astronomy Observatory Key Science Program (TRAO-KSP), "mapping Turbulent properties In star-forming MolEcular clouds down to the Sonic scale" (TIMES; PI: Jeong-Eun Lee). Here, we investigate the characteristic relations between the observed lines by performing the Principal Component Analysis (PCA). We also investigate the correlation between the line intensity distributions and the physical parameters, such as the gas column density and dust temperature. Finally, we will discuss how the correlations among different chemical tracers vary with the star formation environments.

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Inhibition by Imatinib of Expression of O-glycan-related Glycosyltransferases and Tumor-associated Carbohydrate Antigens in the K562 Human Leukemia Cell Line

  • Sun, Qi-Chang;Liu, Mi-Bo;Shen, Hong-Jie;Jiang, Zhi;Xu, Lan;Gao, Li-Ping;Ni, Jian-Long;Wu, Shi-Liang
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.4
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    • pp.2447-2451
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    • 2013
  • Objective: To study changes of tumor associated carbohydrate antigen (TACAs) expression and mRNA levels for tumor associated glycosyltransferases, and assess subcellular localizations of N-acetyl galactosyltransferases (GalNAc-Ts) in the K562 leukemia cell line after imatinib treatment. Methods: RT-PCR was performed to analyze the expression of glycosyltransferases which synthesize O-glycan in tumor-associated carbohydrate antigens (TCTAs). The expression of Tn antigen, T antigen and sialyl T antigen on K562 cell membranes was measured by flow cytometry after treatment with different concentrations of imatinib. Co-localization of GalNAc-Ts and ER (endoplasmic reticulum) was determined by confocal laser scanning microcopy. Results: Transcript expression levels of several glycosyltransferases related to TCTAs were decreased after imatinib ($0-0.3{\mu}M$) treatment. Expression of Tn antigen and T antigen was increased while that of sialyl T antigen was decreased. Co-localization of GalNAc-Ts and ER was reduced by $0.2{\mu}M$ of imatinib. Conclusion: Imatinib inhibited the expression of O-glycan related TACAs and several related glycosyltransferases, while decreasing the co-localization of GalNAc-Ts and ER and normalizing O-glycosylation in the K562 human leukemia cell.

Differential antiangiogenic and anticancer activities of the active metabolites of ginsenoside Rg3

  • Maryam Nakhjavani;Eric Smith;Kenny Yeo;Yoko Tomita;Timothy J. Price;Andrea Yool;Amanda R. Townsend;Jennifer E. Hardingham
    • Journal of Ginseng Research
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    • v.48 no.2
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    • pp.171-180
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    • 2024
  • Background: Epimers of ginsenoside Rg3 (Rg3) have a low bioavailability and are prone to deglycosylation, which produces epimers of ginsenoside Rh2 (S-Rh2 and R-Rh2) and protopanaxadiol (S-PPD and R-PPD). The aim of this study was to compare the efficacy and potency of these molecules as anti-cancer agents. Methods: Crystal violet staining was used to study the anti-proliferatory action of the molecules on a human epithelial breast cancer cell line, MDA-MB-231, and human umbilical vein endothelial cells (HUVEC) and compare their potency. Cell death and cell cycle were studied using flow cytometry and mode of cell death was studied using live cell imaging. Anti-angiogenic effects of the drug were studied using loop formation assay. Molecular docking showed the interaction of these molecules with vascular endothelial growth factor receptor-2 (VEGFR2) and aquaporin (AQP) water channels. VEGF bioassay was used to study the interaction of Rh2 with VEGFR2, in vitro. Results: HUVEC was the more sensitive cell line to the anti-proliferative effects of S-Rh2, S-PPD and R-PPD. The molecules induced necroptosis/necrosis in MDA-MB-231 and apoptosis in HUVEC. S-Rh2 was the most potent inhibitor of loop formation. In silico molecular docking predicted a good binding score between Rh2 or PPD and the ATP-binding pocket of VEGFR2. VEGF bioassay showed that Rh2 was an allosteric modulator of VEGFR2. In addition, SRh2 and PPD had good binding scores with AQP1 and AQP5, both of which play roles in cell migration and proliferation. Conclusion: The combination of these molecules might be responsible for the anti-cancer effects observed by Rg3.

Interaction of HIV-1 Core p24 Antigen with Human Monocytic Cell Line THP1 Results in TNF-${\alpha}$ Dependent Secretion of Matrix Metalloproteinase-9

  • Sung, Ji-Hye;Yoo, Seung-Hee;Park, Hae-Kyung;Chong, Young-Hae
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.9-18
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    • 2000
  • Immunological mechanisms involving the release of inflammatory factors by HIV-1 infected microglia in the brain have been implicated in the pathogenesis of HIV dementia (HIVD). Since the regulation of matrix metalloproteinases (MMPs) activity can be influenced by variety of inflammatory mediators, this study was undertaken to look for a correlation between the MMP-9 release and the production of TNF-${\alpha}$ in response to HIV-1 p24 in the human monocyte cell line THP-1 as a model for microglia. First, it was shown that HIV-l core p24 antigen induced THP-1 to secrete MMP-9 in a dose response manner while it elicited a little effect on MMP-2 release in human astroglial cell line T98G. Next, it was found that p24 induced THP-1 to secrete TNF-${\alpha}$ without prior differentiation into macrophages by phorbol myristate acetate (PMA) treatment. Furthermore, anti-TNF-${\alpha}$ neutralizing antibodies significantly blocked p24-induced MMP-9 release in a dose dependent manner. Our data indicate that p24 antigen induces monocytic MMP-9 release by triggering up-regulation of TNF-${\alpha}$ secretion.

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Gene Expression in Zn-deficient U937 Cell Line : Using cDNA Microarray (아연결핍된 단핵구 U937 Cell Line에 있어서의 유전자 발현 탐색 : cDNA Microarray 기법 이용)

  • Beattie, John H.;Trayhurn, Paul
    • Journal of Nutrition and Health
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    • v.35 no.10
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    • pp.1053-1059
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    • 2002
  • In post-genome period, the technique for identifying gene expression has been changed to high throughput screening. In the field of molecular nutrition, the need for this technique to clarify molecular function of the specific nutrient is essential. In this study, we have tested the zinc-regulated gene expression in zinc-deficient U937 cells, using cDNA microarray which is the cutting-edge technique to screen large numbers of gene expression simultaneously. The study result can be used for the preliminary gene screening data for clarifying, using monocyte U937 cell line, molecular Zn aspect in atherosclerosis. U937 cells were cultured in Zn-adequate (control, 12 $\mu$M Zn) or Zn-deficient (experimental, 0 $\mu$M Zn) ESMI media during 2 days, respectively. Cells were harvested and RNA was extracted. Total RNA was reverse-transcriptinized and synthesized cDNA probe labeled with Cy-3. fluorescent labeled cDNA probe was applied to microarray slide for hybridization slide, and after then, the slide was scanned using fluorescence scanner. ‘Highly expressed genes’ in Zn-deficient U937 cells, comparing to Zn-adequate group, are mainly about the genes for motility protein, immune system protein, oncogene and tumor suppressor and ‘Less highly expressed genes’ are about the genes for transcription, apoptosis associated protein, cell cycle, and several basic transcription factors. The results of this preliminary study imply the effectiveness of cDNA microarray for expression profiling of a singly nutrient deficiency, specially Zn. Furthur study, using tailored-cDNA array and capillary endothelial cell lines, would be beneficial to clarify molecular Zn function, more in detail.

Phenotypic characterization of pre-harvest sprouting resistance mutants generated by the CRISPR/Cas9-geminiviral replicon system in rice

  • Jong Hee Kim;Jihyeon Yu;Jin Young Kim;Yong Jin Park;Sangsu Bae;Kwon Kyoo Kang;Yu Jin Jung
    • BMB Reports
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    • v.57 no.2
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    • pp.79-85
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    • 2024
  • Pre-harvest sprouting is a critical phenomenon involving germination of seeds in the mother plant before harvest under relative humid conditions and reduced dormancy. In this paper, we generated HDR mutant lines with one region SNP (C/T) and an insertion of 6 bp (GGT/GGTGGCGGC) in OsERF1 genes for pre-harvest sprouting (PHS) resistance using CRISPR/Cas9 and a geminiviral replicon system. The incidence of HDR was 2.6% in transformed calli. T1 seeds were harvested from 12 HDR-induced calli and named ERF1-hdr line. Molecular stability, key agronomic properties, physiological properties, and biochemical properties of target genes in the ERF1-hdr line were investigated for three years. The ERF1-hdr line showed significantly enhanced seed dormancy and pre-harvest sprouting resistance. qRT-PCR analysis suggested that enhanced ABA signaling resulted in a stronger phenotype of PHS resistance. These results indicate that efficient HDR can be achieved through SNP/InDel replacement using a single and modular configuration applicable to different rice targets and other crops. This work demonstrates the potential to replace all genes with elite alleles within one generation and greatly expands our ability to improve agriculturally important traits.