• Title/Summary/Keyword: base-isolated

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Genetic Diversity of a Natural Population of Apple stem pitting virus Isolated from Apple in Korea

  • Yoon, Ju Yeon;Joa, Jae Ho;Choi, Kyung San;Do, Ki Seck;Lim, Han Cheol;Chung, Bong Nam
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.195-199
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    • 2014
  • Apple stem pitting virus (ASPV), of the Foveavirus genus in the family Betaflexiviridae, is one of the most common viruses of apple and pear trees. To examine variability of the coat protein (CP) gene from ASPV, eight isolates originating from 251 apple trees, which were collected from 22 apple orchards located in intensive apple growing areas of the North Gyeongsang and North Jeolla Provinces in Korea, were sequenced and compared. The nucleotide sequence identity of the CP gene of eight ASPV isolates ranged from 77.0 to 97.0%, while the amino acid sequence identity ranged from 87.7 to 98.5%. The N-terminal region of the viral CP gene was highly variable, whereas the C-terminal region was conserved. Genetic algorithm recombination detection (GARD) and single breakpoint recombination (SBP) analyses identified base substitutions between eight ASPV isolates at positions 54 and 57 and position 771, respectively. GABranch analysis was used to determine whether the eight isolates evolved due to positive selection. All values in the GABranch analysis showed a ratio of substitution rates at non-synonymous and synonymous sites (dNS/dS) below 1, suggestive of strong negative selection forces during ASPV CP history. Although negative selection dominated CP evolution in the eight ASPV isolates, SLAC and FEL tests identified four possible positive selection sites at codons 10, 22, 102, and 158. This is the first study of the ASPV genome in Korea.

Production of Theileria sergenti recombinant protein by E coli expression system

  • Park, Jin-ho;Chae, Joon-seok;Kim, Dae-hyuk;Jang, Yong-suk;Kwon, Oh-deong;Lee, Joo-mook
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.786-796
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    • 1999
  • As an attempt to develop an effective control method against theileriosis, recombinant antigen protein was produced. Thirty-two kDa membrane protein(MP) gene of T sergenti was amplified through RT-PCR from extracted total RNA of T sergenti isolated in Chonbuk, Korea. The amplified 869 bp of Korean T sergenti membrane gene was cloned and the base sequences were analyzed. The amplified gene was cloned into E coli expression vector, pQE32 plasmid vector, and the vector was introduced into E coli strain M15 to produce the recombinant membrane protein. For the induction of T sergenti membrane protein(KTs-MP), the plasmid harboring E coli strain M15 were cultured in the presence of IPTG, and the recombinant protein were purified by $Ni^+$-NTA agarose. Then, to confirm the authenticity of the produced membrane protein, molecular weight of expressed recombinant KTs-MP was analyzed by SDS-PAGE and Western blotting. The molecular weight of expressed recombinant protein was 32 kDa as expected. The recombinant KTs-MP was successfully recognized by anti-His Tag antibody, antisera of T sergenti infected cattle and monoclonal antibody of T sergenti membrane protein. Therefore, we concluded that the authentic 32 kDa membrane protein of T sergenti was produced as immunologically recognizable form.

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Suppression of Matrix Metalloproteinase-9 Expression of Flavonoids from Metasequoia glyptostroboides (낙우송(Metasequoia glyptostroboides)으로부터 분리한 flavonoid의 금속단백분해효소-9 발현 억제 활성)

  • Yang Jae-Young;Lee Ho-Jae;Kho Yung-Hee;Kwon Byoung-Mok;Chun Hyo Kon
    • Journal of Life Science
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    • v.15 no.2 s.69
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    • pp.231-235
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    • 2005
  • Matrix metalloproteinases (MMPs) are a family of structurally and functionally related zinc-dependent enzymes responsible for proteolytic degradation of extracellular matrix components such as base membrane or interstitial stroma. MMPs play an important role in a variety of physiological and pathological tissue remodeling processes, including wound healing, embryo implantation, tumor invasion and metastasis. Since MMP-9 (gelatinase B) has unique ability to cleave type IV collagen, gene expression of MMP-9 has been focused on as a pharmacological target. Flavonoids are a class of compounds that are widely spread in plants. In the coures of screening for the suppressors of MMP-9 gene expression from natural products, Metasequoia glyptostroboides was selected. Six flavonoids, sciadopitysin, isoginkgetin, bilobetin, 2,3-dihydrohinokiflavone, luteolin and apigenin were purified as suppressors of MMP-9 gene expression from M. glyptostroboides. The suppressing activity of the isolated flavinoids on the MMP-9 gene expression was measured by gelatin zymography and Nothern blot analysis.

Molecular Cloning and Characterization of Sesquiterpene Cyclase cDNAs from Pepper Plant Infected with Phytophthora capsici

  • Kim, Jong-Bum;Lee, Sung-Gon;Ha, Sun-Hwa;Lee, Myung-Chul;Ye, Wan-Hye;Lee, Jang-Yong;Lee, Shin-Woo;Kim, Jung-Bong;Cho, Kang-Jin;Hwang, Young-Soo
    • Journal of Applied Biological Chemistry
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    • v.44 no.2
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    • pp.59-64
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    • 2001
  • Pepper plants (Nogkwang, 60-day old) were inoculated with Phytophthora capsici to induce sesquiterpene cyclase associated with the biosynthesis of phytoalexin (capsidiol), a substance related to the defense against pathogens in plants. One day after inoculation, mRNA was isolated from the root, cDNA synthesized, and a library constructed in a ZAP express XR vector. The efficiency was $2{\times}10^6pfu/{\mu}g$. Sesquiterpene cyclase cDNA from Hyoscyamus muticus was labeled with $^{32}P$ and used as a probe for screening the cDNA library. After the third screening, 25 positive clones were selected. Through restrictive digestion and DNA gel-blot analysis, six different cyclase gene expressions were identified. PSC1B sequences of the six clones were determined, which were 1966 base pairs encoded 556 amino acids with an expected molecular weight of 63.8 kDa. Response against the pathogen was different between the resistant and susceptible peppers. After the infection of the pathogen, the expression of PSC genes continued in the resistant peppers while the plants were alive. The expression in the susceptible peppers lasted for only 4 days.

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Cloning and Nucleotide Sequence Analysis of the asd Gene from Shigella sonnei KNIH104S (Shigella sonnei KNIH104S로부터 asd 유전자의 클로닝 및 염기서열 분석)

  • 박용춘;신희정;김영창
    • Korean Journal of Microbiology
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    • v.35 no.1
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    • pp.13-17
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    • 1999
  • Shigella sonnez is important causes of human enleric infcctions. S. sonnei KNIH104S was isolated from patient of shigellosis in Korea and previously reported. We cloned 1.7 kb BamHI fragment containing the asd gene encoding an aspartate $\beta$-semialdehyde dehydrogenase from chromosomal DNA of S. sonnei KNIH104S. This recombinant plasmid was named as pSAB17. E. coli $\chi$6097, an a d mutant, cannol grow on the LB medium without DL-$\alpha$, $\varepsilon$-diaminopimclic acid (50 pgiml) but E. coli x 6097(pSAB17) can grow on the same medium. We sequenccd the asd gene ol Shigella for the first time. The asd gcne was composed of 1,104 base pairs with ATG initiation codon and TAA termination codon. Sequence comparison of the asd gene exhibited 99.9% nucleolide sequence hornology with that of E. coli. Also, We constructed the balanced-lethal vector using pBluescrip SK(+) and asd gene of S. sonnei KNIH104S.

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Cloning and Nucleotide Sequence Analysis of the aroA Gene from Salmonella typhi KNIH100 (Salmonella typhi KNIH100으로부터 aroA 유전자의 클로닝과 염기서열 분석)

  • 길영식;신희정;김영창
    • Korean Journal of Microbiology
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    • v.36 no.1
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    • pp.46-51
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    • 2000
  • Salmonella typhi is one of important causes of human enteric infections. S. typhi KNIH100 was isolated from a patient of typhoid fever in Korea. We cloned a 5.0 kb SalⅠ fragment containing the aroA gene encoding a 5-enolpyruvylshikimate-3-phosphate synthetase from chromosomal DNA of this strain. This recombinant plasmid was named pSAL80. E. coli CGSC2829, an aroA- mutant, was not grown on the M9 minimal medium but E. coli CGSC2829 (pSAL80) was grown on the M9 minimal medium. The aroA gene was composed of 1,284 base pairs with ATG initiation codon and TAA termination codon. Sequence comparison of the aroA gene exhibited 99%, 98%, and 77% identity with those of S. typhi Ty2, S. typhimurium, and E. coli respectively. As in the cases of Shigella sonnei and E. coli, the serC and aroA genes lie in a single operonic structure.

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Synthesis of Chelating Adsorbent (2,2'-Iminodibenzoic Acid-crosslinked Chitosan) and Adsorptivity of Pb(II), Cu(II), Cd(II) (킬레이트 흡착제(2,2'-Iminodibenzoic acid-가교 chitosan)의 합성과 Pb(II), Cu(II), Cd(II)의 흡착력에 관한 연구)

  • Shim, Sang-Kyun;Ryu, Jae-Jun
    • Analytical Science and Technology
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    • v.11 no.6
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    • pp.452-459
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    • 1998
  • Crosslinked chitin was prepared from epichlorohydrin and chitin which was isolated from waste marin source. The crosslinked chitosan were prepared by the deacetylation of the crosslinked chitin with a strong base. 2,2'-Iminodibenzoic acid-crosslinked chitosan was prepared by reacting 2,2'-Iminodibenzoic acid salt with crosslinked chitosan-Cl which was obtained by chlorination of crosslinked chitosan. The adsorptivity of Pb(II), Cu(II), Cd(II) was studied as a synthetic adsorbent. Experimental results for the adsorption and the recovery characteristics showed that the more pH increase, the more amount of adsorbed metal ion increase. Optimum adsorption time was 1 hr, and adsorption capacity was increased in order of $Cu^{2+}$<$Cd^{2+}$<$Pb^{2+}$, and recovery capacity was increased in order of $Cd^{2+}$<$Cu^{2+}$<$Pb^{2+}$.

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Comparison of antigenicity of Edwardsiella tarda isolates in loach(Misgurnus mizloepis) (미꾸라지에서의 Edwardsiella tarda isolates의 항원성 비교)

  • Lee, Young;Jun, Lyu-Jin;Kim, Myoung-Suk;Park, Kyung-Hyun;Jeong, Hyun-Do
    • Journal of fish pathology
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    • v.21 no.3
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    • pp.201-208
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    • 2008
  • We compared the pathogenicity and antigenicity of two different Edwardsiella tarda (E. tarda) strains KFE and Edk-2 isolated in Korea and Japan respectively. In the pathogenicity analysis with challenge test against loach, E. tarda KFE isolate showed stronger pathogenicity compared to that of E. tarda Edk-2 isolate. The differences were also confirmed by the comparison of OMP (outer membrane protein) in SDSPAGE which showed three major bands, 41kDa, 37kDa and 30kDa, for E. tarda KFE isolates and two major bands, 41kDa and 30kDa, for E. tarda Edk-2 isolates. On the base of these results, we tried to determine the differences of antigenicities of these two isolates in loach which is one of the important species in freshwater aquaculture in Korea. Numbers of specific antibody secreting cells (SASC), appeared to be higher in loach immunized with FKC of E. tarda Edk-2 than loach immunized with FKC of E. tarda KFE. ELISPOT-assay for the comparison of antigenicity showed relatively high percentage of cross-reaction and implied the presence of some common epitopes in the antigens of these two E. tarda isolates.

Characterization of a Novel Alga-Lytic Bacterium, Acidovorax temperans AK-05, Isolated from an Eutrophic Lake for Degradation of Anabaena cylindrica (부영양 호수에서 분리한 Acidovorax temperans AK-05의 Anabaena cylindrica 분해 특성)

  • Kim, Jeong-Dong;Han, Myung-Soo
    • Korean Journal of Ecology and Environment
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    • v.37 no.2 s.107
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    • pp.241-247
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    • 2004
  • Isolation and identification of alga-lytic bacteria were carried out. Fifteen isolates of alga-lytic bacteria were screened by the double layer method using A. cylindrica NIES-19 as a sole nutrient and four isolates among them were compared with their alga-lytic activity. The isolate AK-05 exhibiting the highest alga-lytic activity was identified as Acidovorax temperans base on its 16S rDNA sequence. The culture supernatant of the isolate AK-05 was reliable for the alga-lytic. Alga-lytic activity assays of culture supernatant revealed that the major substances for alga-lytic activity were non-proteins and heat stable. The highest alga-Iytic activity was practical under alkaline conditions and at 25${\sim}$$30^{\circ}C$. It is indicating an advantage for the application of water blooms by cyanobacteria in eutrophic lakes where the pH is generally in alkaline region.

Potentiality of Using Vertical and Three-Dimensional Isolation Systems in Nuclear Structures

  • Zhou, Zhiguang;Wong, Jenna;Mahin, Stephen
    • Nuclear Engineering and Technology
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    • v.48 no.5
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    • pp.1237-1251
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    • 2016
  • Although the horizontal component of an earthquake response can be significantly reduced through the use of conventional seismic isolators, the vertical component of excitation is still transmitted directly into the structure. Records from instrumented structures, and some recent tests and analyses have actually seen increases in vertical responses in base isolated structures under the combined effects of horizontal and vertical ground motions. This issue becomes a great concern to facilities such as a Nuclear Power Plants (NPP), with specialized equipment and machinery that is not only expensive, but critical to safe operation. As such, there is considerable interest worldwide in vertical and three-dimensional (3D) isolation systems. This paper examines several vertical and 3D isolation systems that have been proposed and their potential application to modern nuclear facilities. In particular, a series of case study analyses of a modern NPP model are performed to examine the benefits and challenges associated with 3D isolation compared with horizontal isolation. It was found that compared with the general horizontal isolators, isolators that have vertical frequencies of no more than 3 Hz can effectively reduce the vertical in-structure responses for the studied NPP model. Among the studied cases, the case that has a vertical isolation frequency of 3 Hz is the one that can keep the horizontal period of the isolators as the first period while having the most flexible vertical isolator properties. When the vertical frequency of isolators reduces to 1 Hz, the rocking effect is obvious and rocking restraining devices are necessary.