• Title/Summary/Keyword: Isolation System

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Isolation and molecular identification of Vibrio harveyi from the look down fish (Selene vomer)

  • Jun, Jin-Woo;Lee, Young-Ran;Giri, Sib Sankar;Kim, Hyoun-Joong;Yun, Sae-Kil;Chi, Cheng;Kim, Sang-Guen;Koh, Jeong-Rack;Jung, Ji-Yun;Lee, Byeong-Chun;Park, Se-Chang
    • Korean Journal of Veterinary Research
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    • v.56 no.2
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    • pp.125-127
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    • 2016
  • Gradual mortality of look down fish (Selene vomer) was observed in a private aquarium in Seoul, showing abnormal swimming behavior and lethargy. A bacterial pathogen from kidney was cultured, identified, and confirmed as Vibrio harveyi using Vitek System 2 and 16S rRNA gene sequencing. A predominant bacterial strain, SNUVh-LW2 was proved to be most closely related to isolates from China by phylogenetic analysis with minimum evolution method. Also, tetracycline was considered as the most sensitive antibiotic agent via antibiotic susceptibility test. The group of fish was treated according to the diagnostic result and no more mortality was observed.

Isolation and Characterization of Lactic Acid Bacteria Producing Antimutagenic Substance from Korean Dongchimi (동치미로부터 항돌연변이 물질을 생산하는 유산균의 분리 및 특성)

  • 주길재;이창호;우철주
    • Journal of Life Science
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    • v.11 no.5
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    • pp.432-438
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    • 2001
  • Various lactic acid bacteria were isolated from Korean Dongchimi (whole radish Kimichi with added water) in order to study their antimutagenic activity. Ames test using Salmonella enterica serovar typhimurium TA98 and TA100 showed the strain DLAB19 to have the highest antimutagenic activity among the 300 isolated strains against MNNG(N-methyl-N-nitro-N-nitrosoguanidine), NPD (4-nitro-O-phenylenediamine), 4-NQO(4-nitroquinoline-1-oxide) and AFB$_{1}$(aflatoxin B$_{1}$). The strain was identified as Leuconostoc mesenteroides subsp. cremoris according to the Bergeys Mannual Systematic Bscteriology based on its morphological, cultural, physiological characteristics and biological system Antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was found in the culture supernatant suggesting the bacterium secretes, the antimutagenic substance in the media. The antimutagenic activity of Leu. mesenteroides subsp. cremoris DLAB19 was reconfirmed by the spore-rec assay using spores of Bacillus subtilis H17 (Rec$^{+}$) and M45 (Rec$^{[-10]}$ ).).

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Growth Analysis of Chlamydomonas reinhardtii in Photoautotrophic Culture with Microdroplet Photobioreactor System (미세액적 광생물반응기를 활용한 광독립영양배양에서 Chlamydomonas reinhardtii의 성장성 분석)

  • Sung, Young Joon;Kwak, Ho Seok;Choi, Hong Il;Kim, Jaoon Young Hwan;Sim, Sang Jun
    • Korean Chemical Engineering Research
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    • v.55 no.1
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    • pp.80-85
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    • 2017
  • Recently, microalgae which can produce high-value products have attracted increasing attention for biological conversion of $CO_2$. However, low photosynthetic efficiency and productivity have limited the practical use of microalgae. Thus, we developed microdroplet photobioreactor for the analysis of photoautotrophic growth of model alga, Chlamydomonas reinhardtii. $CO_2$ transfer rate was increased by integrating micropillar arrays and adjusting height of microchamber. These results were identified by change of cell growth rate and fluorescence intensity. Lastly, the photoautotrophic growth kinetics of C. reinhardtii in microdroplet photobioreactor were investigated under different $CO_2$ concentrations and light intensities for 96 hours. As a result, microdroplet photobioreactor was efficient platform for isolation and rapid evaluation of microalgal strains which have enhanced productivity of high-value products and growth performance.

Isolation and Characterization of Glycolate Oxidase Gene from Panax ginseng C. A. Meyer

  • Parvin, Shohana;Pulla, Rama Krishna;Kim, Yu-Jin;Sathiyaraj, Gayathri;Jung, Seok-Kyu;Khorolragchaa, Altanzul;In, Jun-Gyo;Yang, Deok-Chun
    • Journal of Ginseng Research
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    • v.33 no.4
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    • pp.249-255
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    • 2009
  • The oxidation of glycolate to glyoxylate, a key step in plant photorespiration, is carried out by the peroxisomal flavoprotein glycolate oxidase (EC 1.1.3.15). To investigate the altered gene expression and the role of GOX in ginseng plant defense system, a cDNA clone containing a GOX gene designated as PgGOX was isolated and sequenced from Panax ginseng. The cDNA was 692 nucleotides long and have an open reading frame of 552 bp with a deduced amino acid sequence of 183 residues. A GenBank BlastX search revealed that the deduced amino acid of PgGOX shares a high degree homology with the Glycine max (95% identity). In the present study we analyzed the expression of PgGOX under various environmental stresses at different times using real time-PCR. The results showed that the expressions of PgGOX increased after various treatments involving salt, light, cold, ABA, SA, and copper treatment.

The transposition pattern of the Ac element and its use for targeted transposition in Arabidopsis thaliana

  • Machida, Yasunori;Onouchi, Hitoshi;Tanaka, Hirokazu;Hamada, Susumu;Ishikawa, Takaaki;Semiarti, Endang;Iwakawa, Hidekazu;Nomura, Kiyohito;Machida, Chiyoko
    • Proceedings of the Botanical Society of Korea Conference
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    • 1999.07a
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    • pp.11-15
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    • 1999
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1,700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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세포내의 물의 상태

  • 강사욱
    • Proceedings of the Botanical Society of Korea Conference
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    • 1985.08b
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    • pp.51-57
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    • 1985
  • In order to evlauate feasibility of the gene tagging by the maize transposable element Ac in heterologous plant systems, we have investigated physical distances and directions of transposition of the element in Arabidopsis thaliana and tobacco cultured cell line BY-2. We prepared a T-DNA construct that carried a non-autonomous derivative of Ac with a site for cleavage by endonuclease I-Scel (designated dAc-I-RS element). Another cleavage site was also introduced into the T-DNA region outside dAc-I-RS. A number of transgenic Arabidopsis plants were generated, each of which had a single copy of the T-DNA at a different chromosomal location. To examine the pattern of transposition, three out of these transgenic plants were crossed with the Arabidopsis plant that carried the gene for Ac transposase and progeny in which dAc-I-RS had been transposed were isolated. After digestion of the genomic DNA of these progeny with I-SceI, sizes of segment of DNA were determined byd pulse-field gel electrophoresis. We also performed linkage analysis for the transposed elements and sites of mutations near the elements. Our results with three transgenic lines showed that 50% of all transposition events had occurred within 1, 700 kilo-base pairs (kb) on the same chromosome, with 35% within 200 kb, and that the elements transposed in both directions on the chromosome with roughly equal probability. The data thus indicate that the Ac-Ds system is most useful for tagging of genes that are present within 200 kb of the chromosomal site of Ac in Arabidopsis. In addition, determination of the precise localization of the transposed dAc-I-RS element should definitely assist in map-based cloning of genes around insertion sites. In the present paper, we report typical examples of such gene isolation studies.

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A Study on the Molecular Biology of Renin-Angiotensin System : Renin Gene Expression and Construction of Genomic Library (Renin-Angiotensin계의 분자생물학적 연구 : Renin유전자의 발현과 Genomic Library작성)

  • 박영순;한동민;김종호;문영희;이호섭;고건일;김성준
    • The Korean Journal of Zoology
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    • v.33 no.1
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    • pp.35-44
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    • 1990
  • Poly (A) + RNA was isolated from mouse submaxillary gland and renin mRNA was isolated by poly (U)-sepharose chromatography and sucrose linear densiW gradient centifugation. And renin mRNA was identified by in vitro translation and immunoprecipitation. In order to construct recombinant plasmid, renin cDNA was synthesized by using reverse transcriptase and inserted into EcoRi site of PUC19. In addition, the cDNA was also synthesized using polymerase chain reaction and inserted into HindlIl site of PUC19. The recombinant plasmid was transformed into JMlO3 and the expression of the inserted renin cDNA was examined. The transformant produced renin protein having a molecular weight of 45, 000 dolton, which showed hypertensive effect upon injecting it into rabbit ear vein. A renin genomic library was prepared by inserting rabbit kidney DNA into EMBL3 phage, and was screeined for the isolation of renin gemomic DNA using renin cDNA probe.

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Monitoring the Differentiation and Migration Patterns of Neural Cells Derived from Human Embryonic Stem Cells Using a Microfluidic Culture System

  • Lee, Nayeon;Park, Jae Woo;Kim, Hyung Joon;Yeon, Ju Hun;Kwon, Jihye;Ko, Jung Jae;Oh, Seung-Hun;Kim, Hyun Sook;Kim, Aeri;Han, Baek Soo;Lee, Sang Chul;Jeon, Noo Li;Song, Jihwan
    • Molecules and Cells
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    • v.37 no.6
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    • pp.497-502
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    • 2014
  • Microfluidics can provide unique experimental tools to visualize the development of neural structures within a microscale device, which is followed by guidance of neurite growth in the axonal isolation compartment. We utilized microfluidics technology to monitor the differentiation and migration of neural cells derived from human embryonic stem cells (hESCs). We co-cultured hESCs with PA6 stromal cells, and isolated neural rosette-like structures, which subsequently formed neurospheres in suspension culture. Tuj1-positive neural cells, but not nestin-positive neural precursor cells (NPCs), were able to enter the microfluidics grooves (microchannels), suggesting that neural cell-migratory capacity was dependent upon neuronal differentiation stage. We also showed that bundles of axons formed and extended into the microchannels. Taken together, these results demonstrated that microfluidics technology can provide useful tools to study neurite outgrowth and axon guidance of neural cells, which are derived from human embryonic stem cells.

Characteristics of Pichia anomala K15 Producing Killer Toxin Isolated from Traditional Nuruk (전통 누룩으로부터 분리된 Killer Toxin 생산 균주 Pichia anomala K15의 특성)

  • Jung, Hee-Kyoung;Park, Chi-Duck;Lee, Gee-Dong;Park, Seung-Chun;Park, Hwan-Hee;Hong, Joo-Heon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.8
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    • pp.1077-1082
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    • 2007
  • In this study, killer yeasts were isolated from traditional Nuruk to improve storage and suppress contaminant in food industry. Among killer yeasts, yeast K15 showed strong killer toxin activity and inhibited growth of Salmonella Typhimurium and Vibrio parahaemolyticus. Killer yeast K15 was identified with Pichia anomala by the Microlog TM 4.0 identification system and homology of the ITS sequence. Killer toxin generated from P. anomala K15 was inactivated by pronase E and suggested to be a protein. Therefore killer toxin of P. anomala K15 was thought to be safe in human such as bacteriocin. P. anomala K15 was sufficient for growth in 50% glucose and could be used to prevent contaminant in initial stages of alcohol beverage fermentation.

Correlation between Ceria abrasive accumulation on pad surface and Material Removal in Oxide CMP (산화막 CMP에서 세리아 입자의 패드 표면누적과 재료제거 관계)

  • Kim, Young-Jin;Park, Boum-Young;Jeong, Hae-Do
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 2008.06a
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    • pp.118-118
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    • 2008
  • The oxide CMP has been applied to interlayer dielectric(ILD) and shallow trench isolation (STI) in chip fabrication. Recently the slurry used in oxide CMP being changed from silica slurry to ceria (cerium dioxide) slurry particularly in STI CMP, because the material selectivity of ceria slurry is better than material selectivity of silica slurry. Moreover, the ceria slurry has good a planarization efficiency, compared with silica slurry. However ceria abrasives make a material removal rate too high at the region of wafer center. Then we focuses on why profile of material removal rate is convex. The material removal rate sharply increased to 3216 $\AA$/min by $4^{th}$ run without conditioning. After $4^{th}$ run, material removal rate converged. Furthermore, profile became more convex during 12 run. And average material removal rate decreased when conditioning process is added to end of CMP process. This is due to polishing mechanism of ceria. Then the ceria abrasive remains at the pad, in particular remains more at wafer center contacted region of pad. The field emission scanning electron microscopy (FE-SEM) images showed that the pad sample in the wafer center region has a more ceria abrasive than in wafer outer region. The energy dispersive X-ray spectrometer (EDX) verified the result that ceria abrasive is deposited and more at the region of wafer center. Therefore, this result may be expected as ceria abrasives on pad surface causing the convex profile of material removal rate.

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