• Title/Summary/Keyword: GENE FLOW

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Population Genetic Structure of Japanese Anchovy (Engraulis japonicus) in Korean waters Based on Mitochondrial 12S Ribosomal RNA Gene Sequences (미토콘드리아 12S 리보종 RNA 유전자배열에 의한 한국해역 멸치 개체군의 유전자 구조)

  • Kim Jin Yeong;Cho Eun Seob;Kim Woo Jin
    • Journal of Life Science
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    • v.14 no.6 s.67
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    • pp.938-950
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    • 2004
  • We used portions of mitochondrial 125 ribosomal RNA gene sequences (339 bp) to investigate the phylogenetic and population genetic characteristics of the Japanese anchovy, Engraulis japonicus, in Korean waters. A total of 35 mtDNA haplotypes were obtained from the samples collected in 3 locations (the southern area of the Yellow Sea, the western coast of Jejudo, and the eastern area of the South Sea) in Korean waters. One haplotype, AN8T103, obtained from the southern area of the Yellow Sea, was formed according to an independent phylogenetic individual in the PAUP analysis, which was separated from the others by a $0.2-4.1\%$ sequence divergence. This distinct haplotype appeared to be one that was carried by immigrants from another study area, but further study is necessary. Genetic divergence, except for AN8T103, was moderate to substantial $(0.2-3.8\%)$ and nucleotide diversity within populations was 0.015 for Yellow Sea, 0.013 for Jejudo, and 0.D15 for South Sea, respectively. The female gene flow was substantial or high (Nm=25.5-36.4), and the genetic distances between regions were not statistically significant $(P>0.01)$. These results indicated that the Japanese anchovy populations occurring in Korean waters were consisted of individuals randomly dispersed over geographic areas.

Effects of tktA, $aroF^{FBR}$and aroL Expression in the Tryptophan-Producing Escherichia coli

  • Kim, Tae-Hyun;Namgoong, Suk;Kwak, Joon-Hyeok;Lee, Se-Yong;Lee, Heung-Shick
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.789-796
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    • 2000
  • In order to analyze the effects of tktA, $aroF^{FBR}$, and aroL expression in a tryptophan-producing Escherichia coli, a series of plasmids carrying the genes were constructed. Introduction of tktA, $aroF^{FBR}$, and aroL into the E. coli strain resulted in approximately 10-20 fold increase in the activities of transketolase, the feedback inhibition-resistant 3-deoxy-D-arabinoheptulsonate-7-phosphate synthase, and shikimate kinase. Expression of $aroF^{FBR}$ in the aroB mutant strain of E. coli resulted in the accumulation of 10 mM of 3-deoxy-D-arabinoheptulsonate-7-phosphate (DAHP) in the medium. Simultaneous expression of tktA and $aroF^{FBR}$ in the strain further increased the amount of excreted DAHP to 20 mM. In contrast, the mutant strain which has no gene introduced accumulated 0.5 mM of DAHP. However, the expression of tktA and $aroF^{FBR}$ in a tryptophan-producing E. coli strain did not lead to the increased production of tryptophan, but instead, a significant amount of shikimate, which is an intermediate in the tryptophan biosynthetic pathway, was excreted to the growth medium. Despite the fact that additional expression of shikimate kinase in the strain could possibly remove 90% of excreted shikimate to 0.1 mM, the amount of tryptophan produced was still unchanged. Removing shikimate using a cloned aroL gene caused the excretion of glutamate, which suggests disturbed central carbon metabolism. However, when cultivated in a complex medium, the strain expressing tktA, $aroF^{FBR}$, and aroL produced more tryptophan than the parental strain. These data indicate that additional rate-limiting steps are present in the tryptophan biosynthetic pathway, and the carbon flow to the terminal pathway is strictly regulated. Expressing tktA in E. coli cells appeared to impose a great metabolic burden to the cells as evidenced by retarded cell growth in the defined medium. Recombinant E. coli strains harboring plasmids which carry the tktA gene showed a tendency to segregate their plasmids almost completely within 24h.

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Over-expression of PTEN Involved in Troglitazone-induced Apoptosis in Human Osteosarcoma Cells (사람골육종세포주의 트로글리타존 유도 세포사에서 PTEN의 역할)

  • Yoon, Sun-Jung;Zhou, Lu;Kim, Jung-Ryul
    • The Journal of the Korean bone and joint tumor society
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    • v.17 no.1
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    • pp.23-29
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    • 2011
  • Purpose: We investigated the effects of phosphatase and tensin homologue deleted on chromosome 10 gene phosphatase and tensin homologue deleted on chromosome 10 gene (PTEN) expression on the cell proliferation and on the responsiveness of troglitazone in osteosarcoma cells. Materials and Methods: Western blotting alnalysis was performed to detect the expression of PTEN in U-2OS cells treated with troglitazone. WST (water-soluble tetrazolium) assay was used to evaluate cell proliferation. Flow cytometry was used to determine cell apoptosis. Further, transfection of wild-type PTEN plasmid DNA was used to upregulate PTEN expression. Results: Troglitazone treatment induced growth inhibition of U2-OS cells in a dose- and time-dependent manner. Troglitazone increased the expression of PTEN in a dose-dependent manner. PTEN upregulation induced by troglitazone treatment resulted in cell growth inhibition and apoptosis in U-2OS cells. PTEN over-expression by plasmid transfection enhanced these effects of troglitazone. Moreover, no changes were observed in the mutant type-PTEN group. Conclusion: Upregulation of PTEN is involved in the inhibition of cell growth and induction of cell apoptosis by troglitazone. Further, PTEN over-expression can cause cell growth inhibition in osteosarcoma cells and these cell growth inhibitions could be enhance by troglitazone treatment.

Interleukin-7 Enhances the in Vivo Anti-tumor Activity of Tumor-reactive CD8+ T cells with Induction of IFN-gamma in a Murine Breast Cancer Model

  • Yuan, Chun-Hui;Yang, Xue-Qin;Zhu, Cheng-Liang;Liu, Shao-Ping;Wang, Bi-Cheng;Wang, Fu-Bing
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.265-271
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    • 2014
  • Interleukin-7 (IL-7) is a potent anti-apoptotic cytokine that enhances immune effector cell functions and is essential for lymphocyte survival. While it known to induce differentiation and proliferation in some haematological malignancies, including certain types of leukaemias and lymphomas, little is known about its role in solid tumours, including breast cancer. In the current study, we investigated whether IL-7 could enhance the in vivo antitumor activity of tumor-reactive $CD8^+$ T cells with induction of IFN-${\gamma}$ in a murine breast cancer model. Human IL-7 cDNA was constructed into the eukaryotic expression plasmid pcDNA3.1, and then the recombinational pcDNA3.1-IL-7 was intratumorally injected in the TM40D BALB/C mouse graft model. Serum and intracellular IFN-${\gamma}$ levels were measured by ELISA and flow cytometry, respectively. $CD8^+$ T cell-mediated cytotoxicity was analyzed using the MTT method. Our results showed that IL-7 administration significantly inhibited tumor growth from day 15 after direct intratumoral injection of pcDNA3.1-IL-7. The anti-tumor effect correlated with a marked increase in the level of IFN-${\gamma}$ and breast cancer cells-specific CTL cytotoxicity. In vitro cytotoxicity assays showed that IL-7-treatment could augment cytolytic activity of $CD8^+$ T cells from tumor bearing mice, while anti-IFN-${\gamma}$ blocked the function of $CD8^+$ T cells, suggesting that IFN-${\gamma}$ mediated the cytolytic activity of $CD8^+$ T cells. Furthermore, in vivo neutralization of $CD8^+$ T lymphocytes by CD8 antibodies reversed the antitumor benefit of IL-7. Thus, we demonstrated that IL-7 exerts anti-tumor activity mainly through activating $CD8^+$ T cells and stimulating them to secrete IFN-${\gamma}$ in a murine breast tumor model. Based on these results, our study points to a potential novel way to treat breast cancer and may have important implications for clinical immunotherapy.

Synthesis and Evaluation of F-18 Labeled 2'-Deoxy-2'-fluoro-5-methyl-1-β-L-arabinofuranosyluracil (L-[18F]FMAU)

  • Jo, Nam-Hyun;Moon, Byung-Seok;Hong, Su-Hee;An, Gwang-Il;Choi, Tae-Hyun;Cheon, Gi-Jeong;Cho, Jung-Hyuck;Yoo, Kyung-Ho;Lee, Kyo-Chul;Oh, Chang-Hyun
    • Bulletin of the Korean Chemical Society
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    • v.28 no.12
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    • pp.2449-2453
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    • 2007
  • L-[18F]FMAU ([18F]1b) was prepared from the precursor 2-O-[(trifluoromethyl)-sulfonyl]-1,3,5-tri-Obenzoyl- α-L-ribofuranose, by coupling the radioactive fluoro-sugar with the corresponding silylated thymine in 4 steps. The final products, including the α and β anomers, were purified using reverse phase HPLC with an appropriate solvent (5% CH3CN/H2O) at a flow rate of 3.0 mL/min. The total elapsed time of synthesis was about 180-200 min from EOB. The α/β anomeric ratio of the compounds was about 1:9, and the radiochemical purity of the product (β-form) was >98% with decay-corrected yields of 25-35%. All radioactive samples were confirmed using co-injection with pure non-radioactive analogues in every step. In the cellular uptake in vitro test of herpes simplex virus-thymidine kinase (HSV1-TK) gene expressed cells, the percent uptake of injected dose (%ID) of L- and D-FMAU was 37.28 and 65.86, respectively after 240 min incubation. However, the relative uptake (MCA-TK/MCA cellular uptake ratio) of L-FMAU was higher than that of D-FMAU (%ID of L-FMAU, 0.36 and D-FMAU, 0.93 after 240 min incubation in MCA cells). This means that L-FMAU will show better specific HSV1-TK gene expressed cell uptake for selective HSV1-TK gene imaging.

MiR-26a promotes apoptosis of porcine granulosa cells by targeting the 3β-hydroxysteroid-Δ24-reductase gene

  • Zhang, Xiaodong;Tao, Qiangqiang;Shang, Jinnan;Xu, Yiliang;Zhang, Liang;Ma, Yingchun;Zhu, Weihua;Yang, Min;Ding, Yueyun;Yin, Zongjun
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.4
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    • pp.547-555
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    • 2020
  • Objective: Apoptosis of ovarian granulosa cells (GCs) affects mammalian follicular development and fecundity. This study aimed to explore the regulatory relationship between microRNA-26a (miR-26a) and the 3β-hydroxysteroid-Δ24-reductase gene (DHCR24) gene in porcine follicular granular cells (pGCs), and to provide empirical data for the development of methods to improve the reproductive capacity of pigs. Methods: The pGCs were transfected with miR-26a mimic, miR-26a inhibitor and DHCR24-siRNA in vitro. The cell apoptosis rate of pGCs was detected by the flow cytometry. The secretion levels of estradiol (E2) and progesterone (P) in pGCs were detected by enzyme-linked immunosorbent assay. Double luciferase validation system was used to detect the binding sites between miR-26a and DHCR24 3'-UTR region. Qualitative real-time polymerase chain reaction and Western blotting were used to verify the DHCR24 mRNA and protein expression in pGCs, respectively, after transfecting with miR-26a mimic and miR-26a inhibitor. Results: Results showed that enhancement of miR-26a promoted apoptosis, and inhibited E2 and P secretion in pGCs. Meanwhile, inhibition of DHCR24 also upregulated the Caspase-3 expression, reduced the BCL-2 expression, promoted pGCs apoptosis, and inhibited E2 and P secretion in pGCs. There were the binding sites of miR-26a located within DHCR24 3'-UTR. Up-regulation of miR-26a inhibited DHCR24 mRNA and protein expression in pGCs. Conclusion: This study demonstrates that miR-26a can promote cell apoptosis and inhibit E2 and P secretion by inhibiting the expression of DHCR24 in pGCs.

Population Genetic Structure of the Korean Endemic Species, Iksookimia pacifica (Pisces: Cobitidae) Distributed in Northeast Korea (한국고유종 북방종개(어류강, 미꾸리과)의 집단유전학적 구조)

  • Jang, Sook-Jin;Ko, Myeong-Hun;Kwan, Ye-seul;Won, Yong-Jin
    • Korean Journal of Environment and Ecology
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    • v.31 no.5
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    • pp.461-471
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    • 2017
  • Population genetic studies of 10 groups of Iksookimia pacifica were conducted to investigate the genetic diversity and population genetic structure across its known range in South Korea. Population DNA sequences of one mitochondrial gene (mtCOI) and three nuclear genes (IRBP, EGR2B, RAG1) were examined in samples collected from ten streams that flow into the East Sea. Both mitochondrial and nuclear sequences exhibited significant differentiation among populations except a few cases. The Bayesian analysis of the multi-locus genotypes inferred from the DNA sequences of nuclear genes clustered the individual fish largely into two geographical groups: a northern group (from Baebong stream to Cheonjin stream) and a southern group (Yangyangnamdae stream to Gangneungnamdae stream). Given that the streams flowing into the East Sea are geographically isolated water systems, such separation of genotypes can be interpreted by the geographical separation of common ancestors into north and south that had colonized South Korea. Since the initial geographical separation of the ancestral population by north and south, the ancestral groups seem to have experienced further differentiation into the current genetic clusters through the physical isolation of streams by the East Sea in each region. It is notable that many individuals in the Jasan stream formed a genetic cluster with those of Yangyangnamdae and Gangneungnamdae streams which are distant from each other. In addition, mitochondrial gene showed low genetic differentiation between some neighboring populations and very low level of genetic diversity in several populations. The present population genetic study will provide valuable information for the conservation and management of the Korean endemic fish species, I. paicifica.

Characterization of the Alzheimer's disease-related network based on the dynamic network approach (동적인 개념을 적용한 알츠하이머 질병 네트워크의 특성 분석)

  • Kim, Man-Sun;Kim, Jeong-Rae
    • Journal of the Korean Institute of Intelligent Systems
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    • v.25 no.6
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    • pp.529-535
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    • 2015
  • Biological networks have been handled with the static concept. However, life phenomena in cells occur depending on the cellular state and the external environment, and only a few proteins and their interactions are selectively activated. Therefore, we should adopt the dynamic network concept that the structure of a biological network varies along the flow of time. This concept is effective to analyze the progressive transition of the disease. In this paper, we applied the proposed method to Alzheimer's disease to analyze the structural and functional characteristics of the disease network. Using gene expression data and protein-protein interaction data, we constructed the sub-networks in accordance with the progress of disease (normal, early, middle and late). Based on this, we analyzed structural properties of the network. Furthermore, we found module structures in the network to analyze the functional properties of the sub-networks using the gene ontology analysis (GO). As a result, it was shown that the functional characteristics of the dynamics network is well compatible with the stage of the disease which shows that it can be used to describe important biological events of the disease. Via the proposed approach, it is possible to observe the molecular network change involved in the disease progression which is not generally investigated, and to understand the pathogenesis and progression mechanism of the disease at a molecular level.

Study on the Runoff Characteristics of Non-point Source Pollution in Municipal Area Using SWMM Model -A Case Study in Jeonju City (SWMM모델을 이용한 도시지역 비점오염원의 유출특성 연구 -전주시를 대상으로)

  • Paik Do-Hyson;Lim Young-Hwan;Choi Jin-Kyu;Jung Paul-Gene;Kwak Dong-Heui
    • Journal of Environmental Science International
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    • v.14 no.12
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    • pp.1185-1194
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    • 2005
  • The runoff characteristics of non-point source pollutions in the municipal area of Jeonju were investigated and analyzed by using the SWMM (Storm Water Management Model). The flow rates and water qualities of runoff from two types of drainage conduits were measured respectively. One was a conventional combined sewer system and the other was a separated sewer system constructed recently From August to November in 2004, investigations on two rainfall events were performed and flow rate, pH, BOD, COD, SS, T-N and T-P were measured. These data were also used for model calibration. On the basis of the measured data and the simulation results by SWMM, it is reported that $80-90\%$ of pollution load is discharged in the early-stage storm runoff. Therefore, initial 10-30 mm of rainfall should be controlled effectively for the optimal treatment of non-point source pollution in urban area. Also, it was shown that the SWMM model was suitable for the management of non-point source pollution in the urban area and for the analysis of runoff characteristics of pollutant loads.

Verification of Hovering Rotor Analysis Code Using Overlapped Grid (중첩격자를 이용한 제자리비행 로터 해석 코드의 수치특성)

  • Kim, Jee-Woong;Park, Soo-Hyung;Yu, Yung-Hoon;Kim, Eu-Gene;Kwon, Jang-Hyuk
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.36 no.8
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    • pp.719-727
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    • 2008
  • A 3-D compressible Navier-Stokes solver using overlapped grids is developed to predict a flow-field around a hovering rotor. The flow solver is verified by a parametric study with the grid spacing of wake grid, spatial accuracy and turbulence model. Computations are performed with different Chimera grid systems. Computational results are compared with the experimental data of Caradonna et al. for both blade loading and the tip vortex behavior. Numerical results show good agreements with experiments for the distribution of surface pressure and tip vortex behavior. Pressure distributions over the blade have marginal differences for different numerical methods, whereas large discrepancies are seen in the prediction of the wake behavior. Results unexpectedly show that the vortex strength from an automated cut-paste Chimera grid is weaker than that from the conventional Chimera grid.