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Development of Multifunctional Microorganisms for the Effective Wastewater Treatment by Synthetic Detergent Decomposing Microorganisms in Wastewater (폐하수중 합성세제분해균을 이용한 다처리기능균주(多處理技能菌株)의 개발(開發))

  • Heo, Jong-Soo;Cho, Ju-Sik;Lee, Hong-Jae;Han, Mun-Gyu;Lim, Yeong-Sung;Ha, Yeong-Lae
    • Korean Journal of Environmental Agriculture
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    • v.12 no.3
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    • pp.230-238
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    • 1993
  • To develop multifunctional microorganisms for effective wastewater treatment, the cell of P. aeruginosa P1 enable to accumulate lead in its cell were conjugated with the cell of P. fluorescens S1 enable to degrade efficiently synthetic detergents. The plasmids of the P. aeruginosa P1 and the P. fluorescens S1 were found in the cell of the conjugants when determined by agarose gel electrophoresis. The conjugants obtained from P. fluorescens S1 as a recipient cell and P. aeruginosa P1 as a donor cell possessed the ability to degrade synthetic detergents as well as to accumulate lead.

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Genetic Transformation of Biocontrol Agent Bacillus sp, YBL-7 by Plasmid pE194 (생물방제균 Bacillus sp. YBL-7의 형질전환조건)

  • 한길환;정병곤;김상달
    • Microbiology and Biotechnology Letters
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    • v.20 no.4
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    • pp.384-389
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    • 1992
  • Bacillus sp. YBL-7 which had been isolated from ginseng root-rot suppressive soil was able to antagonize Fusarium solani causing ginseng root-rot by their antibiotic substance. In order to develop multifunctional antagonist on Bacillus sp. YBL-7 as a biocontrol agent against Fusarium salam', optimal conditions for protoplast transformation system of Bacillus sp.YBL-7 by the vector plasmid pE194 were investigated. The protoplasts of Bacillus sp. YBL-7 were obtained at best efficiency by treatment with 200${\mu}g$/ml of lysozyme in the pH 7.0 of SMM buffer for 90 minutes at $40^{\circ}C$. The cell wall of the protoplast was regenerated on the agar plate containing 1.2% agar and 0.7 M mannitol. Under the best condition for protoplast formation and regeneration, the optimal transformation was achieved with 40% polyethylene glycol (M.W. 4000) treatment for 10minutes. The vector plasmid pE194 showed the best transformation frequency at 5$\mu$g/ml of final concentration. The pE194 was very stable over 80% in the transformants.

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The Application of Single Nucleotide Polymorphism Markers for Discrimination of Sweet Persimmon Cultivars (단감 품종 판별을 위한 single nucleotide polymorphism 마커 적용 검정)

  • Park, Yeo Ok;Choi, Seong-Tae;Son, Ji-Young;Kim, Eun-Gyeong;Ahn, Gwang-Hwan;Park, Ji Hae;Joung, Wan-Kyu;Jang, Young Ho;Kim, Dong Wan
    • Journal of Life Science
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    • v.30 no.7
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    • pp.614-624
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    • 2020
  • The recent development of next-generation sequencing technology has enabled increased genomic analysis, but very few single nucleotide polymorphism (SNP) markers applicable to sweet persimmon (Diospyros kaki Thunb.) cultivars have been identified. In this study, SNP primers developed from five pollination-constant astringent (PCA) persimmons native to Korea were applied to discriminate between cultivars and verify their usability. The polymerase chain reactions of 19 SNP primers developed by Jung et al. were checked, with 11 primers finally selected. The other eight were very difficult to analyze in the agarose gel electrophoresis and QIAxcel Advanced System used in this experiment and were therefore excluded. The 11 SNP primers were applied through first and second verification to 76 cultivars and collection lines including 20 pollination-variant non-astringent (PVNA), 30 pollination-constant non-astringent (PCNA), 20 PCA, and six pollination-variant astringent (PVA). Of these, 38 were indistinguishable (eight PVNA, 18 PCNA, nine PCA, and three PVA). However, the results of applying the 11 SNP primers to new sweet persimmon cultivars, namely Gamnuri, Dannuri, Hongchoo, Jamisi, and Migamjosaeng, showed that they have the potential to be used as a unique marker for simultaneously determining between them.

Expression of angiogenin, TGF-${\beta}$, VEGF, APEX and TNF-${\alpha}$ in oral squamous cell carcinoma

  • Lee, Ho-Sun;Kim, Kyoung-Won;Kim, Wun-Jae
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.32 no.1
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    • pp.8-18
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    • 2006
  • Purpose: The purpose of this study was to verify that the expressions of angiogenin, transforming growth factor-beta(TGF-${\beta}$), vascular endothelial growth factor(VEGF), human apurinic/apyrimidinic endonuclease(APEX) and tumor necrosis factor-alpha(TNF-${\alpha}$) were associated with the tumorigenesis of the oral squamous cell carcinoma(OSCC). Materials and Methods: Fifty-one samples of OSCC and fifteen normal oral mucosae were obtained to analyze the expression levels of above five factors. mRNA expressions were quantified by the quantitative competitive PCR(QC-PCR) method. After 2% agarose gel electrophoresis stained with ethidium bromide, the concentration of mRNA was calculated by a digital image analysis system. The expression levels of angiogenin, TGF-${\beta}$, VEGF, APEX and TNF-${\alpha}$ were compared by unpaired Student's t-tests between cancer and normal tissues. We analyzed statistically to find the cut-off values that would be useful as diagnostic markers, and the linear regression analysis between every two factors of these five factors by SAS system. Results: All of these five factors (angiogenin: P<0.0037, TGF-${\beta}$: P<0.0001, VEGF: P<0.0102, APEX: P<0.0023, TNF-${\alpha}$: P<0.0074) were significantly correlated with OSCC. In the analysis to find the cut-off values for the diagnosis, we could not find any value that had a reasonable sensitivity and specificity. In the linear regression analysis, there were correlations between angiogenin and TNF-${\alpha}$, TGF-${\beta}$ and VEGF, TGF-${\beta}$ and APEX, TGF-${\beta}$ and TNF-${\alpha}$, VEGF and APEX, VEGF and TNF-${\alpha}$, APEX and TNF-${\alpha}$. Conclusion: Our results suggest that not only angiogenin, TGF-${\beta}$, VEGF, APEX and TNF-${\alpha}$ are significantly associated with the tumorigenesis, but also the close relationship between these factors might enhance the tumorigenesis of OSCC. We can not find clinical availability for diagnosis.

Apoptotic Cell Death of Human Leukemia U937 Cells by Essential Oil purified from Schisandrae Semen (오미자 종자 정유에 의한 인체백혈병 U937 세포의 apoptosis 유도)

  • Choi, Yung Hyun
    • Journal of Life Science
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    • v.25 no.2
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    • pp.249-255
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    • 2015
  • Schisandrae fructus [Schizandra chinensis (Turcz.) Baillon] is a medicinal herb widely used for treating various inflammatory and immune diseases in East Asian countries. The Schisandrae Semen essential oil (SSeo) from this plant has pharmacological activities, including antioxidant, antimicrobial, and antitumoral activities. Nevertheless, the biological activities and underlying molecular mechanisms of the potential anti-cancer effects of this oil remain unclear. In the present study, we investigated the potential inhibition of apoptosis signaling pathways by SSeo in human leukemia U937 cells and evaluated the underlying molecular mechanism. Exposure to SSeo resulted in a concentration-dependent growth inhibition due to apoptosis, which was verified by DNA fragmentation, the presence of apoptotic bodies, and an increase in the sub-G1 ratio. Induction of apoptotic cell death by SSeo was correlated with the down-regulation of members of the inhibitor of apoptosis protein (IAP) family (including X-linked inhibitor of apoptosis protein (XIAP), cIAP-1, and surviving) and anti-apoptotic Bcl-2, and with up-regulation of death receptor (DR) 4 and DR5, depending on dosage. SSeo treatment also induced Bid truncation, mitochondrial dysfunction, proteolytic activation of caspase-3, -8 and -9, and concomitant degradation of activated caspase-3 target proteins such as poly (ADP-ribose) polymerase. Taken together, these findings suggest that SSeo may be a potential chemotherapeutic agent for use in the control of human leukemia cells. Further studies are needed to identify its active compounds.

Induction of Apoptosis by HDAC Inhibitor Trichostatin A through Activation of Caspases and NF-κB in Human Prostate Epithelial Cells. (인체 전립선 상피세포에서 HDAC 저해제 trichostatin A의 caspase 및 NF-κB의 활성화를 통한 apoptosis 유도)

  • Park, Cheol;Jin, Cheng-Yun;Choi, Byung-Tae;Lee, Won-Ho;Choi, Yung-Hyun
    • Journal of Life Science
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    • v.18 no.3
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    • pp.336-343
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    • 2008
  • Histone deacetylases (HDACs) inhibitors have emerged as the accessory therapeutic agents for various human cancers, since they can block the activity of specific HDACs, restore the expression of some tumor suppressor genes and induce cell differentiation, cell cycle arrest and apoptosis in vitro and in vivo. In the present study, we investigated that the effect of trichostatin A (TSA), an HDAC inhibitor, on the cell growth and apoptosis, and its effect on the nuclear factor-kappaB $(NF-{\kappa}B)$ activity in 267B1 human prostate epithelial cells. Exposure of 267B1 cells to TSA resulted in growth inhibition and apoptosis induction in and dose-dependent manners as measured by fluorescence microscopy, agarose gel electrophoresis and flow cytometry analysis. TSA treatment inhibited the levels of IAP family members such as c-IAP-1 and c-IAP-2 and induced the proteolytic activation of caspase-3, -8 and -9, which were associated with concomitant degradation of poly (ADP-ribose)-polymerase, ${\beta}-catenin$ and laminin B proteins. The increase in apoptosis by TSA was connected with the translocation of $NF-{\kappa}B$ from cytosol to nucleus, increase of the DNA binding as well as promoter activity of $NF-{\kappa}B$, and degradation of cytosolic inhibitor of KappaB $(I{\kappa}B)-{\alpha}$ protein. We therefore concluded that TSA demonstrated anti-proliferative and apoptosis-inducing effects on 267B1 cells in vitro, and that the activation of caspases and $NF-{\kappa}B$ may play important roles in its mechanism of action. Although further studies are needed, these findings provided important insights into the possible molecular mechanisms of the anti-cancer activity of TSA.

Rapid and Specific Identification of Genus Cynoglossus by Multiplex PCR Assays Using Species-specific Derived from the COI Region (다중 PCR 분석법을 이용한 참서대과 어종의 신속하고 정확한 종판별 분석법 개발)

  • Noh, Eun Soo;Kang, Hyun Sook;An, Cheul Min;Park, Jung Youn;Kim, Eun Mi;Kang, Jung Ha
    • Journal of Life Science
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    • v.26 no.9
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    • pp.1007-1014
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    • 2016
  • A highly efficient, rapid, and reliable multiplex polymerase chain reaction based method for distinguishing ten species of genus Cynoglossus (C. senegalensis, C. abbreviates, C. macrolepidotus, C. arel, C. semilaevis, C. interruptus, C. joyneri, C. lingua, C. robustus, and C. monodi) is described. The species-specific primer sets were designed base on the cytochrome oxidase subunit I gene (1,500 bp). The optimal PCR conditions and primers were selected for ten of Cynoglossus species to determine target base sequences using single PCR. Multiplex PCR using the ten pairs of primers either specifically amplified a DNA fragment of a unique size or failed, depending on each species DNA. The length of amplification fragment of 208 bp for C. senegalensis, 322 bp for C. abbreviates, 493 bp for C. macrolepidotus, 754 bp for C. arel, 874 bp for C. semilaevis, 952 bp for C. interruptus, 1,084 bp for C. joyneri, 1,198 bp for C. lingua, 1,307 bp for C. robustus, and 1,483 bp for C. monodi with the species-specific primers, visualized by agarose gel electrophoresis, allowed perfectly distinction of the Cynoglossus species. The multiplex PCR assay can be easily performed on multiple samples and attain final results in less than 6 hours. This technique should be a useful addition to the molecular typing tools for the tentative identification of Cynoglossus species.

Studies of Insecticide Resistance in Green Peach Aphids, Myzus persicae(Sulz) III. Acephate Resistance, Cross-Resistance, and Esterase Isozymes (복숭아혹진딧물의 살충제 저항성(抵抗性)에 관한 연구(硏究) III. Acephate저항성(抵抗性) 발달(發達), 교차저항성(交叉抵抗性) 및 Esterase Isozymes)

  • Choi, Seung-Yoon;Kim, Gil-Hah
    • Korean journal of applied entomology
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    • v.25 no.2 s.67
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    • pp.99-105
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    • 1986
  • The green peach aphids(Myzus persicae) collected in a field had been successively selected by acephate(O, S-dimethyl N-acetyl phosphoroamidothioate) in the laboratory. The selected aphid strain in the 20th generation demonstrated relatively high resistance to acephate as well as relatively high cross-resistance to cypermethrin and oxydemeton-methyl, except pirimicarb. The different esterase isozymes with the strains were detected by the agarose gel electrophoresis and among the isozymes the band of ${\beta}-2$ was only specific for the acephate resistant strains.

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Differential Expression of Glucose Transporter Gene in Mouse Early Embryos (생쥐 초기배아의 Glucose Transporter유전자 발현 양상에 관한 연구)

  • Youm, Hye-Won;Byun, Hye-Kyung;Song, Gyun-Ji;Kim, Hae-Kwon;Lee, Ho-Joon
    • Clinical and Experimental Reproductive Medicine
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    • v.25 no.1
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    • pp.77-86
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    • 1998
  • The uptake of glucose for metabolism and growth is essential to most animal cells and is mediated by glucose-transporter (GLUT) proteins. The aim of this study was to determine which class of glucose transporter molecules was responsible for uptake of glucose in the mouse early embryo and at which stage the corresponding genes were expressed. In addition, co-culture system with vero cell was used to investigate the effect of the system on GLUT expression. Two-cell stage embryos were collected from the superovulated ICR female and divided into 3 groups. As a control, embryos were cultured in 0.4% BSA-T6 medium which includes glucose. For the experimental groups, embryos were cultured in either co-culture system with vero cells or glucose-free T6 medium supplemented with 0.4% BSA and pyruvate as an energy substrate. 2-cell to blastocyst stage embryos in those groups were respectively collected into microtubes (50 embryos/tube). Total RNA was extracted and RT-PCR was performed. The products were analysed after staining ethidium bromide by 2% agarose gel electrophoresis. Blastocysts were collected from each group at l20hr after hCG injection. They were fixed in 2.5% glutaraldehyde, stained with hoechst, and mounted for observation. In control, GLUT1 was expressed from 4-cell to blastocyst. GLUT2 and GLUT3 were expressed in morula and blastocyst. GLUT4 was expressed in all stages. When embryos were cultured in glucose-free medium, no significant difference was shown in the expression of GLUT1, 2 and 3, compared to control. However GLUT4 was not expressed until morular stage. When embryos were co-cultured with vero cell, there was no significant difference in the expression of GLUT1, 2, 3 and 4 compared to control. To determine cell growth of embryos, the average cell number of blastocyst was counted. The cell number of co-culture ($93.8{\pm}3.1$, n=35) is significantly higher than that of control and glucose-free group ($76.6{\pm}3.8$, n=35 and $68.2{\pm}4.3$, n=30). This study shows that the GLUT genes are expressed differently according to embryo stage. GLUTs were detectable throughout mouse preimplantation development in control and co-culture groups. However, GLUT4 was not detected from 2- to 8-cell stage but detected from morula stage in glucose-free medium, suggested that GLUT genes are expressed autocrinally in the embryo regardless of the presence of glucose as an energy substrate. In addition, co-culture system can increase the cell count of blastocyst but not improve the expression of GLUT. In conclusion, expression of GLUT is dependent on embryo stage in preimplantation embryo development.

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Identification of Korean Native Goat Meat using DNA Analysis (DNA분석기법을 이용한 한국재래산양육의 판별)

  • Sang, B.C.;Lee, S.H.;Ryoo, S.H.;Seo, K.W.;Han, S.W.;Kim, S.K.
    • Korean Journal of Agricultural Science
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    • v.26 no.2
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    • pp.33-38
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    • 1999
  • This study was carried out to analyze the genetic polymorphisms of genomic DNA of blood and meat for conservation of the genetic resources and genetic improvement of Korean Native goat. The genetic identification between Korean Native goat and imported goat was examined using RAPD(random amplified polymorphisms DNAs) analysis with 30 Korean Native goat, 10 hybrid, 10 imported goat. 10 Korean native goat meat and 10 imported goat meat. The results obtained from this study were summarized as follows: 1. Genomic DNA from Korean native goat, hybrid and imported goat could be obtained above about 23kb size using 0.5% agarose gel electrophoresis and the ratio of optical density at 260nm to that at 280nm was between 1.7 and 2.0 using UV spectrophtometer instrument. 2. In the results of the gene identification between Korean Native goat and hybrid, and imported goat using RAPD methods with random primer of 110 kinds, only Korean native goat showed a specific band at about 369bp using a random primer OPO-19 (5'-CAA ACG TCG G-3'), but imported goat and hybrid not showed. 3. Also, in the results of the gene identification between Korean Native goat meat and imported goat meat using RAPD methods with random primer, Korean native goat only showed a specific band at about 369bp using a random primer No. 19(5'-CAA ACG TCG G-3'), but imported goat not showed.

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