• Title/Summary/Keyword: endonuclease

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Circular Plasmid DNA from a Red Algae, Porphyra tenera (양식 참김(Porphyra tenera)에서 분리한 Circular Plasmid DNA)

  • 류태형;최학선;최경희;이춘환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.27 no.6
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    • pp.1160-1165
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    • 1998
  • When total cellular DNA was isolated from Porphyra tenera by ultracentrifugation on Hoechst dye/CsCl gradients method, plasmid like DNA's were concentrated at the upper band which were characterized with a A+T rich organelle DNA's in the CsCl gradients. Based on their electrophoretic migration in different concentration of agarose gel, buffer system, and electric power etc. and the results of restriction digestion, the plasmid like DNA's were concluded to have circular conformation. This is the first report of putative circular plasmid DNA from the P. tenera, which is a autonomously replicating plasmid existing with a high copy number plasmid in the cell. The minimum size of this plasmid estimated by restriction endonuclease digestion was appeared to be 2.5kb in size.

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Production of Nuclease Activity in U937 Cells by Phorbol 12-Myristate 13-Acetate and Lipopolysaccharide

  • Kwon, Hyung-Joo;Kim, Doo-Sik
    • BMB Reports
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    • v.36 no.5
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    • pp.520-523
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    • 2003
  • The proliferation and differentiation signals of myelogeneous U937 cells are provided by extracellular stimuli, such as lipopolysaccharide (LPS) and phorbol 12-myristate 13-acetate (PMA). In a DNA-native-polyacrylamide gel assay system, we demonstrated that a particular nuclease activity is expressed in PMA-stimulated U937 cells and secreted into the culture medium. The nuclease activity was induced in U937 cells by LPS treatment, while the secretion of the enzyme was undetected in the culture medium. Therefore, it is likely that the expression and secretion of the particular nuclease in U937 cells are controlled by extracellular stimulations, such as PMA and LPS treatment.

Characteristics of Bradyrhizobium japonicum SNU001, aSsymbiotic Strain of Glycion max (콩(Glycine max)의 공생균주 Bradyrhizobium japonicum SNU001의 특성)

  • 고세리;박용근;안정선
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.143-147
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    • 1991
  • The root nodules and Glycine max were classified as determinate nodule based on their morphological characteristics, and isolated endosymbiont as a Bradyrhizobium based on its growth rate and single subpolar flagellum. The isolate was similar to B. japonicum USDA110 in utilization of carbon source, growth at 38.deg.C and 2% NaCl, production of $H_{2}$S and especially in the restriction endonuclease digestion pattern of symbiotic genes, allowing them to be placed in sTI group together. The former, however, grew better than the later in broad pH range from 5.0 to 9.5. Infectivity and effectivity of the isolate were confirmed by inoculation of soybean seedlings with the isolates. Characteristics of the reisolated endosymbiont from induced root nodules were identical to those of the first isolate. From these results, it was confirmed that Bradyrhizobium strain isolated from the root nodules of Glycine max was a real symbiont, and was named B. japonicum SNU001.

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한국형 B형 간염 바이러스 elongated X 단백질의 기능 및 간암 유발 기작에 관한 연구 (I)

  • 노현모
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.265-265
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    • 1994
  • 본 연구는 간염 바이러스의 X 및 elongated X 유전자를 클로닝하여 E. coli에서 대량 발현시진 후, 그 기능을 여러 측면에서 연구하교 지금까지 알려진 oncogene products, tumor suppressor, 그리고 그 밖의 다른 암 유발인자와의 interaction에 대해 분석함으로써 간암 생성의 분자적 기작을 이해하고 더 나아가 간암의 예방 및 치료제의 개발을 목표로 하였다. 그 일차적 연구로서 이전에 플로닝된 mutant hepatitis Bvirus genome으로부터 X 및 elongated X 유전자를 클로닝하였으며, E. coli에서 대량 발현시키기 위하여 T7 bacteriophage promoter아래에 재 클로닝하였다. 이러한 X 및 ebngated X 유전자를 E. coli에서 대량 발현시킨 후, rabbit anti-X antibody를 이용하여 western blotting을 수행함으로서 이를 확인하였으며 DEAE-cellulose와 heparin-agarose chromategraphy를 이용하여 순수분리하였다. 순수분리된 X 및 etongated X 단백질을 highly differentiated hepatoma cell인 HepG$_2$ cell에 처리하여 transactivation activity를 측정하였다. 그 결과 순수분리된 단백질들이 SV4O promoter를 transactivation 함을 할 수 있었으며, 이로부터 클로닝된 유전자들이 모두 정상적인 기능을 가짐을 확인하였다. 그러고 X 유전자의 작용기작을 규명하기위하여 restriction endonuclease를 이용하여 5 개의 mutant X 유전자를 구성하였으며 현재 이를 HepG2 cell에 transfection 하여 그 기능을 연구하고 있다.

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Comparison in Restriction Profile Analysis of Vibrio furnissi, Vibrio fluvialis, and Vibrio parahaemolyicus Bacteriophage from Sea Product

  • Younghee Kim
    • Journal of Environmental Science International
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    • v.1 no.2
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    • pp.99-103
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    • 1992
  • The bacteriophages lytic for Vibrio furnissi, Vibrio furniulis and Vibrio parahemolyticus were isolated from fish gills and shellfish. Nucleic acid of bacteriophage was prepared and restriction endonuclease profile was compared. All isolates contained deoxyribonucleic acid. V. fumissi bacteriophage from fish gills showed 2 bands with Bgl II, 1 with Pst, 3 with Hind III, 1 with Bm HI and 2 with EcoR I. V Puuialis phage represented 7 fragments with Bgl II, 1 with Pst, 4 with Hind III, and 2 with EcoR I. V parhemolyticn produced 13 sites with Hind III and 4 sites with EcoR I. The fragment types were varied depending on the phage isolation. All three phages were digested with Hind III and EcoR I with different sizes. V furnissi phage were digested with 5 different restriction enzymes. Key words: Bacteriophage, Vibrio furnissi, Vibrio fluvialis, Vibrio pnrahemolyticus, Deoxyribonucleic acid, Pst, Bam HI, Hind III, EcoR I, Bgl II.

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Analysis of Tetracycline Resistance Plasmids and the Conjugative Transfer of Tetracycline Resistancy in Staphylococcus epidermidis (Staphylococcus epidermidis의 Tetracycline 내성 Plasmid의 분석 및 Conjugation에 의한 내성전달)

  • Chung, Jae-Kyu;Park, Mi-Kyung;Kim, Sung-Kwang
    • The Journal of the Korean Society for Microbiology
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    • v.22 no.1
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    • pp.95-99
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    • 1987
  • When tetracycline resistancy were cured by ethidium bromide treatment, some of the cured strains lost the tetracycline resistance plasmid while other strains kept the plasmids. Both strains of lost and remained plasmids were digested with restriction endonuclease Hind III and these cleaved plasmids were compared with that of parent strains, two plasmid remained strains showed same cleavage patterns between parent and cured strains, however, one plasmid lost strain showed dissimilarity with parent strain, but in the other one strain, among 4 plasmid lost colonies, 2 showed same but other 2 showed different patterns compared to parent strain. Tetracycline was transfered by conjugation in on set(Staphylococcus aureus donor versus Staphylococcus epidermidis, recipient) with relative high frequency but the other 2 sets showed a low degree of frequency and the other 2 sets exhibited no transfer.

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Characterization of Endogeneous Plasmids from Two Bacillus Isolates (Bacillus 속 분리균 2종의 내재형 Plasmids 특성분석)

  • 윤기홍
    • Microbiology and Biotechnology Letters
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    • v.27 no.5
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    • pp.364-369
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    • 1999
  • In order to obtain the suitable plasmids for constructing plasmid vectors of Bacillus species, endogeneous plasmid DNAs were screended from thermo-tolerant soil bacteria. Based on agarose gel electrophoresis patterns of the isolated plasmid DNAs, two strains harboring small-size plasmids were selected. The isolated were identified to belong to the genus Bacillus on the basis of their morphological and biochemical properties, and named Bacillus sp. 3-3 and 77-8, respectively. The restriction endonuclease maps were determined for four plasmids including two plasmids from each Bacillus isolates. It is interesting that Bacillus sp. 3-3 and 77-8 have an identical plasmid according to the restriction maps. The three kinds of hybrid plasmids constructed by introducing each plasmid of two isolates into a Escherichia coli plasmid vector. pUCCm18 containing chloramplenicol resistance gene active in Bacillus strains, could be replicated in B. subtilis and B. licheniformis. These plasmids are very stable in B. subtilis, suggesting that the Bacillus plasmids identified in this work would be useful for development of new cloning vectors for Bacillus strains.

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Comparison of Biotyping, Serotyping and Molecular Typing of Yersinia enterocolitica Isolated from Spring water in Seoul (서울시내 약수에서 분리한 Yersinia enterocolitica의 생물형, 혈청형 및 분자학적 형별비교)

  • 이영기;최성민;오수경;신재영
    • Journal of environmental and Sanitary engineering
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    • v.14 no.4
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    • pp.99-109
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    • 1999
  • Enteropathogenic Yersina enterocolitica is an important cause of human and animal disease. Phenotypic and genotypic characteristics currently used to identify Yersinia enterocolitica are not necessarily sufficient to differentiate pathogenic from non-pathogenic strains or to analyze the epidemiology of yersiniae at a molecular level. To improve the characterization of Yersinia enterocolitica, A total of 65 isolates of Yersinia enterocolitica were examined with bioserotyping, antibiotic susceptibilities, PFGE, PCR-ribotyping. Genomic DNA pattern generated by PFGE are highly specific for different strains of an organism and have significant value in epidemiologic investigations. The PFGE analysis of Not I-digested chromosomal DNA of Y. enterocolitica were performed with a CHEF Mapper(Bio-Rad, USA). Not I generated 19 restriction endonuclease digestion profiles(REDP). PCR-ribotyping, performed with primers complementry to conserved regions of 16S and 23S rRNA gene, generated 13 ribotypes. PCR-ribotyping can be considered a good technich for subtyping strains of Y.enterocolitica.

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CRISPR and Target-Specific DNA Endonucleases for Efficient DNA Knock-in in Eukaryotic Genomes

  • Lee, Seung Hwan;Kim, Sunghyun;Hur, Junho K
    • Molecules and Cells
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    • v.41 no.11
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    • pp.943-952
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    • 2018
  • The discovery and mechanistic understanding of target-specific genome engineering technologies has led to extremely effective and specific genome editing in higher organisms. Target-specific genetic modification technology is expected to have a leading position in future gene therapy development, and has a ripple effect on various basic and applied studies. However, several problems remain and hinder efficient and specific editing of target genomic loci. The issues are particularly critical in precise targeted insertion of external DNA sequences into genomes. Here, we discuss some recent efforts to overcome such problems and present a perspective of future genome editing technologies.

Characterization of Tetracycline Resistant Plasmid in Staphylococcus aureus by Restriction Enzyme Mapping (황색포도상구균에서 테트라사이클린 내성을 나타내는 플라스미드의 동정)

  • Kim, Ki-Hyun;Kim, Jong-Myung;Moon, Kyung-Ho
    • YAKHAK HOEJI
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    • v.36 no.3
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    • pp.255-258
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    • 1992
  • The clinical isolate Staphylococcus aureus SA8 was resistant to tetracycline(Tc) and harboured a plasmid pKH1(24.82 kb). pKH1 was shown by curing and by transformation to specify resistance to Tc. The cleavage map of a pKH1 was determined by restricction enzyme mapping techniques. Cleavage map is given for BglII, EcoRI, HpaII, PvuII and SalI. Restriction endonuclease BamHI, BglI, BstEII, HpaI, PstI, and XhoI have no sites on this plasmid. HaeIII, XbaI, and HindIII have 5, 6, 14 sites, respectively.

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