• Title/Summary/Keyword: Gel electrophoresis

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Microbial Communities in Rice Paddy Soils Following Cultivation of Genetically Modified Leaf Folder-resistant Rice Plants (혹명나방 저항성벼 재배 논토양의 미생물상)

  • Kwon, Jang-Sik;Noh, Hyung-Jun;Suh, Jang-Sun;Shin, Kong-Sik;Kweon, Soon-Jong
    • Korean Journal of Soil Science and Fertilizer
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    • v.43 no.2
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    • pp.180-187
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    • 2010
  • The study was performed to investigate the property of rhizosphere microorganisms, and community structure during GMO, and Non-GMO rice cultivation. In the dilution plate technique, there were no significant differences in microbial populations of rhizosplane with genetically modified, and non-genetically modified rice cultivation, and rhizosphere were also the same results. Dominant bacterial genera were Afipia 12.5%, Spingomonas 10.0%, Ramlibacter 10.0%, Mycobacterium 7.5%, and Tetrasphaera 7.5% in rhizosphere soil of genetically modified rice plant, while Afipia 7.3%, Spingomonas 12.2%, Ramlibacter 7.3%, Mycobacterium 17.1%, Tetrasphaera 14.6% in non-genetically modified cultivated at Suwon test fields in 2006. Majorgenera isolated from root surface cultivated in Yesan fields were Arthrobacter 12.7% in rhizoplane of genetically modified plant, and Burkholderia 22.2% of non-genetically modified plant in 2007, Paucimonas 26.6% of genetically modified plant, Chryseobacterium 15.4% of non-genetically modified plant in 2008. Also the microbial communities in rhizosphere soils of genetically modified, and non-genetically modified plants were characterized using phospholipid fatty acid, and denaturing gradient gel electrophoresis. The phospholipid fatty acid profiles of soils in this condition showed different pattern, but did not show significant differences between soils cultivated with genetically or non-genetically modified rice plants.

Potential Importance of Proteomics in Research of Reproductive Biology (생식생물학에세 프로테오믹스의 응용)

  • Kim Ho-Seung;Yoon Yong-Dal
    • Development and Reproduction
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    • v.8 no.1
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    • pp.1-9
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    • 2004
  • The potential importance of proteomic approaches has been clearly demonstrated in other fields of human medical research, including liver and heart disease and certain forms of cancer. However, reproductive researches have been applied to proteomics poorly. Proteomics can be defined as the systematic analysis of proteins for their identity, quantity, and function. It could increase the predictability of early drug development and identify non-invasive biomarkers of toxicity or efficacy. Proteome analysis is most commonly accomplished by the combination of two-dimensional gel electrophoresis(2DE) and MALDI-TOF(matrix-assisted laser desorption ionization-time of flight) MS(mass spectrometry) or protein chip array and SELDI-TOF(surface-enhanced laser desorption ionization-time of flight) MS. In addition understanding the possessing knowledge of the developing biomarkers used to assess reproductive biology will also be essential components relevant to the topic of reproduction. The continued integration of proteomic and genomic data will have a fundamental impact on our understanding of the normal functioning of cells and organisms and will give insights into complex cellular processes and disease and provides new opportunities for the development of diagnostics and therapeutics. The challenge to researchers in the field of reproduction is to harness this new technology as well as others that are available to a greater extent than at present as they have considerable potential to greatly improve our understanding of the molecular aspects of reproduction both in health and disease.

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Helicobacter pylori Strain 51 (Korean Isolate): Ordered Overlapping BAC Library, Combined Physical and Genetic Map, and Comparative Analysis with H. pylori Strain 26695 and Strain J99

  • KANG HYUNG-LYUN;LEE WOO-KON;SONG JAE-YOUNG;CHOI SANG-HAENG;PARK SEONG-GYU;RYU BOK-DEOK;LEE EUN-JOO;KIM JI-SUN;PARK JEONG-UCK;BAIK SEUNG-CHUL;CHOI MYOUNG-BUM;YOUN HEE-SHANG;KO GYUNG-HYUCK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.844-854
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    • 2005
  • We constructed a defined physical and genetic map of H. pylori strain 51, previously isolated from a Korean patient with a duodenal ulcer, by combining a restriction analysis by pulse-field gel electrophoresis with the construction of a BAC library. A Notl-digest of H. pylori strain 51 genome yielded seven fragments, from which the genomic size was estimated to be 1,698$\pm$24 kb. The BAC library was constructed from 50 to 200 kb fragments of HindIII-digested genomic DNA. From 700 BAC clones, an ordered overlapping maxi-set of 82 BAC clones was assembled that covered the entire genome. The positions of 15 genes were localized in the strain 51 genome with 4-22 kb of resolution and were compared with their orthologues in strain 26695 and strain J99. The arrangement of the 15 genes was identical in strain 51 and strain J99, except for flaA and hpaA. The plasticity zone of strain 51, like that of strain J99, was located in the single region, and was shorter than those of strain 26695 and strain J99. The strain 51 plasticity zone consisted of ORFs common only to strain 51 and J99 or to strain 51 and 26695, as well as strain 51-specific ORFs. Three genetic translocations and/or inversions were found between orthologue ORFs in strain 51 and strain J99. These results show that the chromosomal organization of strain 51 differs from Western strains such as strain 26695 and strain J99.

Optimal Conditions for the Laccase Production from Fomitopsis pinicola Mycelia (Fomitopsis pinicola 균사체로부터 Laccase의 최적생산조건)

  • Park, Naomi;Park, Sang-Shin
    • Microbiology and Biotechnology Letters
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    • v.37 no.1
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    • pp.62-68
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    • 2009
  • The culture conditions to maximize the production of laccase (EC 1.10.3.2) from Fomitopsis pinicola mycelia were investigated. Among the tested media for the enzyme production, mushroom complete medium (MCM ; 2% dextrose, 0.2% peptone, 0.2% yeast extract, 0.05% $KH_2PO_4$, and 0.05% $MgSO_4{\cdot}7H_2O$) showed the highest activity of the enzyme. To optimize the culture condition for the laccase activity, influence of various carbon and nitrogen sources was investigated in MCM. Among various carbon and nitrogen sources, 2% glucose and 0.4% peptone showed the highest production of the enzyme, respectively. For the phosphorus and inorganic source, 0.05% $NaH_2PO_4$ and 0.05% $CaCl_2$ were best for the enzyme activity. The enzyme production was reached to highest level after the cultivation for 8 days at $25^{\circ}C$. Native polyacrylamide gel electrophoresis (PAGE) followed by the laccase activity staining using 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the laccase under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with molecular mass of 52 kDa. The optimum pH and temperature for the enzyme activity were $80^{\circ}C$ and pH 3.0.

Proteomic Profiles of Mouse Neuro N2a Cells Infected with Variant Virulence f Rabies Viruses

  • Wang, Xiaohu;Zhang, Shoufeng;Sun, Chenglong;Yuan, Zi-Guo;Wu, Xianfu;Wang, Dongxia;Ding, Zhuang;Hu, Rongliang
    • Journal of Microbiology and Biotechnology
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    • v.21 no.4
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    • pp.366-373
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    • 2011
  • We characterized the proteomes of murine N2a cells following infection with three rabies virus (RV) strains, characterized by distinct virulence phenotypes (i.e., virulent BD06, fixed CVS-11, and attenuated SRV9 strains), and identified 35 changes to protein expression using two-dimensional gel electrophoresis in whole-cell lysates. The annotated functions of these proteins are involved in various cytoskeletal, signal transduction, stress response, and metabolic processes. Specifically, a-enolase, prx-4, vimentin, cytokine-induced apoptosis inhibitor 1 (CIAPIN1) and prx-6 were significantly up-regulated, whereas Trx like-1 and galectin-1 were down-regulated following infection of N2a cells with all three rabies virus strains. However, comparing expressions of all 35 proteins affected between BD06-, CVS-11-, and SRV9-infected cells, specific changes in expression were also observed. The up-regulation of vimentin, CIAPIN1, prx-4, and 14-3-3 ${\theta}/{\delta}$, and down-regulation of NDPK-B and HSP-1 with CVS and SRV9 infection were ${\geq}2$ times greater than with BD06. Meanwhile, Zfp12 protein, splicing factor, and arginine/serine-rich 1 were unaltered in the cells infected with BD06 and CVS-11, but were up-regulated in the group infected with SRV9. The proteomic alterations described here may suggest that these changes to protein expression correlate with the rabies virus' adaptability and virulence in N2a cells, and hence provides new clues as to the response of N2a host cells to rabies virus infections, and may also aid in uncovering new pathways in these cells that are involved in rabies infections. Further characterization of the functions of the affected proteins may contribute to our understanding of the mechanisms of RV infection and pathogenesis.

Purification and properties of a basic inducible protein, ICG with chitinase and ${\beta}-1,3-glucanase$ activities from rice cell suspension culture media treated with chitooligosaccharides (Chitooligosaccharides 처리에 의해 유도되는 chitinase, ${\beta}-1,3-glucanase$ 활성 보유 벼 염기성 단백질 ICG의 분리 및 성질)

  • Um, Sung-Yon;Park, Hee-Young;Kim, Su-Il
    • Applied Biological Chemistry
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    • v.37 no.1
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    • pp.43-48
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    • 1994
  • A basic inducible protein, ICG, containing chitinase and ${\beta}-1,3-glucanase$ activity concomittantly was purified from cell suspension culture media of rice after the treatment of chitooligosaccharides. The isolated ICG enzyme gave a single band on native and SDS polyacrylamide gel electrophoresis and its molecular weight was estimated to be 52.53 kd. The optimal temperature and optimal pH of both enzyme activities in ICG were $60^{\circ}C$, pH 6.0 for chitinase activity and $37^{\circ}C$, pH 4.0 for ${\beta}-1,3-glucanase$ activity. $K_M$ and $V_{max}$ values for chitinase were 0.474 mM. 2.997 nM/min., and those for ${\beta}-1,3-glucanase$ were 1.004 mM 0.739 nM/min. respectively. TLC analysis of the chitooligosaccharide hydrolysates with ICG enzyme indicated that ICG acts as endochitinase.

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Purification of Xylogone sphaerospora ${\beta}$-mannanase and Growth Activity of Bifidobacterium spp. by Konjac Glucomannan Hydrolysates (Xylogone sphaerospora 유래 ${\beta}$-mannanase 정제 및 Konjac Glucomannan 가수분해 올리고당의 중합도별 Bifidobacterium spp.에 대한 증식활성)

  • Lee, Hee-Jung;Park, Gwi-Gun
    • Applied Biological Chemistry
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    • v.51 no.3
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    • pp.159-163
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    • 2008
  • Xylogone sphaerospora ${\beta}$-mannanase was purified by Sephadex G-100 column chromatography. The specific activity of the purified enzyme was 8.44 units/ml protein, representing an 56.27-folds purification of the original crude extract. The final preparation thus obtained showed a single band on SDS-polyacrylamide gel electrophoresis. The molecular weight was determined to be 42kDa. Konjac glucomannan was hydrolyzed by the purified ${\beta}$-mannanase, and then the hydrolysates was separated by activated carbon column chromatography. The main hydrolysates were composed of D.P. (Degree of Polymerization) 3 and 4 glucomannooligosaccharides. For elucidate the structure of D.P 3 and 4 glucomannooligosaccharides, sequential enzymatic action was performed. D.P 3 and 4 were identified as M-G-M and M-M-G-M (G- and M- represent glucosidic and mannosidic link-ages). To investigate the effects of konjac glucomannooligosaccharides on in vitro growth of Bifido-bacterium longum, B. bifidum, B. infantis, B. adolescentis, B. animalis, B. auglutum and B. breve. Bifidobacterium spp. were cultivated individually on the modified-MRS medium containing carbon source such as D.P. 3 and D.P. 4 glucomannooligosaccharides, respectively. B. longum and B. bifidum grew up 3.9-fold and 2.8-fold more effectively by the treatment of D.P. 4 glucomannooligosaccharides, compared to those of standard MRS medium. Especially, D.P. 4 was more effective than D.P. 3 glucomannooligosaccharide on the growth of Bifidobacterium spp.

Impact of Virus-resistant Trigonal Cactus Cultivation on Soil Microbial Community (바이러스저항성 삼각주 재배가 토양 미생물상에 미치는 영향)

  • Oh, Sung-Dug;Kim, Jong-Bum;Lee, Jung-Jin;Kim, Min-Kyeong;Ahn, Byung-Ohg;Sohn, Soo-In;Park, Jong-Sug;Ryu, Tae-Hun;Cho, Hyun-Suk;Lee, Kijong
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.148-154
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    • 2013
  • BACKGROUND: Genetically modified(GM) trigonal cactus(Hylocereus trigonus Saff.) contained a coat protein gene of cactus virus X (CVX), which conferred resistance to the virus, phosphinothricin acetyltransferase (bar) gene, which conferred herbicide resistance, and a cauliflower mosaic virus 35S promoter (CaMV 35S). This study was conducted to evaluate the possible impact of GM trigonal cactus cultivation on the soil microbial community. METHODS AND RESULTS: Microorganisms were isolated from the rhizosphere of GM and non-GM trigonal cactus cultivation soils. The total numbers of bacteria, and actinomycete in the rhizosphere soils cultivated GM and non-GM trigonal cactus were similar to each other, and there was no significant difference. Dominant bacterial phyla in the rhizosphere soils cultivated with GM and non-GM trigonal cactus were Proteobacteria, Uncultured archaeon, and Uncultured bacterium. The denaturing gradient gel electrophoresis (DGGE) profiles show a similar patterns, significant difference was not observed in each other. DNA was isolated from soil cultivated GM and non-GM trigonal cactus, we analyzed the persistence of the inserted gene by PCR. Amplification of the inserted genes was not observed in the soil DNA, which was collected after harvest. CONCLUSION(S): This result suggests that the GM trigonal cactus cultivation does not change significantly the microbial community.

Effects of Protox Herbicide Tolerance Rice Cultivation on Microbial Community in Paddy Soil (Protox 제초제저항성 벼 재배가 토양미생물 군집에 미치는 영향)

  • Oh, Sung-Dug;Ahn, Byung-Ohg;Kim, Min-Kyeong;Sohn, Soo-In;Ryu, Tae-Hun;Cho, Hyun-Suk;Kim, Chang-Gi;Back, Kyoung-Whan;Lee, Kijong
    • Korean Journal of Environmental Agriculture
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    • v.32 no.2
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    • pp.95-101
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    • 2013
  • BACKGROUND: Rice (Oryza sativa) is the most important staple food of over half the world's population. This study was conducted to evaluate the possible impact of transgenic rice cultivation on the soil microbial community. METHODS AND RESULTS: Microorganisms were isolated from the rhizosphere of GM and non-GM rice cultivation soils. Microbial community was identified based on the culture-dependent and molecular biology methods. The total numbers of bacteria, fungi, and actinomycete in the rhizosphere soils cultivated with GM and non-GM rice were similar to each other, and there was no significant difference between GM and non-GM rice. Dominant bacterial phyla in the rhizosphere soils cultivated with GM and non-GM rice were Actinobacteria, Firmicutes, and Proteobacteria. The microbial communities in GM and non-GM rice cultivated soils were characterized using the denaturing gradient gel electrophoresis (DGGE). The DGGE profiles showed similar patterns, but didn't show significant difference to each other. DNAs were isolated from soils cultivating GM and non-GM rice and analyzed for persistence of inserted gene in the soil by using PCR. The PCR analysis revealed that there were no amplified protox gene in soil DNA. CONCLUSION(S): These data suggest that transgenic rice does not have a significant impact on soil microbial communities, although continued research may be necessary.

Study on the Standardization of a Surveillance Culture Laboratory in Infection Control Fields (감염관리 분야에서 감시배양검사의 표준화 연구)

  • Park, Chang-Eun;Jeong, Na-Yeon;Yang, Min-Ji;Kim, Han-Wool;Joo, Sei-Ick;Kim, Keon-Han;Seong, Hee-Kyung;Hwang, Yu-Yean;Lim, Hyun-Mi;Son, Jae-Cheol;Yoon, Sun-Han;Yoon, Nam-Seob;Jang, In-Ho
    • Korean Journal of Clinical Laboratory Science
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    • v.50 no.3
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    • pp.359-369
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    • 2018
  • An essential measure to prevent healthcare-associated infections (HAI) is to develop a consistent system of surveillance, thereby promoting a reliable situation diagnosis to perform efficient control for the problem. Patient-to-patient transmission of pathogens within the hospital plays a substantial role in the epidemiology of HAIs. Contamination of healthcare environments commonly occurs, including facilities surfaces (e.g., bed rails, bedside tables), drinking water, cooling tower water, endoscopic instruments, food, airborne, endotoxin test, sterile test and medical equipment, with pathogenic organisms. In addition, epidemiological analysis is performed by multi locus sequence tying, pulsed-field gel electrophoresis for active surveillance. Therefore, an environmental surveillance culture test for prevention improves patient safety and blocks infection agents. Effective infection control and increased safety are possible by controlling the national infection control system. In conclusion, this study contributes to an effective infection control system through the standardization of active surveillance culture laboratory and secure expertise as infection control specialist. The primary objective of the standardization is to improve the safety of the nation's healthcare system by reducing the rates of HAIs.