• Title/Summary/Keyword: restriction endonuclease digestion

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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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The Characterization of Mitochondrial DNA of Korean Ginseng (Panax ginseng C.A. Meyer) (고려인삼의 미토콘드리아 DNA의 분자생물학적 특성연구)

  • Lim, Yong-Pyo;Park, Kwang-Tae
    • Journal of Ginseng Research
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    • v.14 no.2
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    • pp.310-316
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    • 1990
  • This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular 9enetical approach of energy Production related mechanism in Panax ginseng. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. mtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not. Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRII treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN I and EcoRII. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector. Genomic library was screened and purified for further research including restriction mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned from the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

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Intraspecific Variation of Environmental and Clinical Vibrio vulnificus Isolates as Demonstrated by Restriction Endonuclease Digestion Profiles

  • Kim, Ki-Yong;Yang, Ho-Chul;Tamplin, Mark-L.;Choi, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.9 no.1
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    • pp.78-83
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    • 1999
  • Thirty-six environmental isolates of Vibrio vulnificus obtained from seawater, sediments, and raw seafoods, and 18 clinical isolates from Vibrio septicemia patients were typed by restriction endonuclease digestion profiles (REDP) of genomic DNA with SfiI. The results revealed a high-level of variation in REDPs, indicating a vast genomic diversity among V. vulnificus strains. Genetic relatedness of the strains showed similarities ranging from 10% to 100%. Different REDPs for isolates from various raw seafoods were obtained, and clustering of strains according to type of seafoods was not observed. In contrast, clinical isolates of V. vulnificus showed higher similarity to one another, and could be subdivided into one separate group. The difference in REDPs of the V. vulnificus isolates from clinical origin and from raw seafoods substantiates the previous observation that only a single type of pathogenic strain was involved in each human infection, despite the numerous genetically polymorphic strains found from implicated oysters.

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Joint Interactions of SSB with RecA Protein on Single-Stranded DNA

  • Kim, Jong-Il
    • Journal of Microbiology and Biotechnology
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    • v.9 no.5
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    • pp.562-567
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    • 1999
  • Single-stranded DNA binding protein (SSB) is well-characterized as having a helix-destabilizing activity. The helix-destabilizing capability of SSB has been re-examined in this study. The results of restriction endonuclease protection assays and titration experiments suggest that the stimulatory effect of SSB on strand exchange acts by melting out the secondary structure which is inaccessible to RecA protein binding; however, SSB is excluded from regions of secondary structure present in native single-stranded DNA. Complexes of SSB and RecA protein are required for eliminating the secondary structure barriers under optimal conditions for strand exchange.

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Synthesis and Characterization of Dodecanucleotides Containing the XhoI Recognition Sequence with a Phosphorothioate Group at the Cleavege Site

  • 문병조;김상국;김남희;권오신
    • Bulletin of the Korean Chemical Society
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    • v.17 no.11
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    • pp.1031-1036
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    • 1996
  • The synthesis and characterization of diastereomeric dodecanucleotides, d[GATCp(S)TCGAGATC], containing recognition sequence of the XhoI restriction endonuclease with a phosphorothioate internucleotidic linkage the cleavage site are described. Rp and Sp form of diastereomerically pure dinucleoside phosphorothioates d[Cp(S)T] were presynthesized and used for the addition to the growing oligonucleotide chain as a block. The stereochemistry of dinucleoside phosphorothioate was assigned by 31P NMR spectroscopy, enzyme digestion, and reverse-phase HPLC. XhoI restriction endonuclease cut only Rp diastereomer d[GATCp(S))TCGAGATC]. The rate of hydrolysis is slower than that of the unmodified dodecamer d[GATCTCGAGATC]. The phosphorothioate nucleotide is using for determination of the stereochemical course of the XhoI catalyzed reaction.

The Characterization of Mitochondrial DNA of Korean Ginseng (Panax ginseng C.A. Meyer)

  • Lim, Yong-Pyo;Park, Kwang-Tae
    • Proceedings of the Ginseng society Conference
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    • 1990.06a
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    • pp.168-174
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    • 1990
  • This study was focused on the characterization of mitochondrial DNA (mtDNA) for molecular genetically approach of energy Production related mechanism in Panax Ein.fend. The simple and efficient method of mtDNA isolation from ginseng has been developed by modification of recently advanced methods. This procedure can successfully apply to mtDNA isolation of several plants. MtDNA of etiolated shoot and one-year root were digested with restriction endonucleases, but that of 6-year root not Any difference was not observed in the restriction endonuclease digestion patterns among the ginseng variants. Molecular size of ginseng mtDNA was estimated at least 159 kb by the restriction endonuclease fragment analysis. The 4.5 kb extra band at the lane of EcoRll treatment could be observed in restriction patterns digested with the methylation sensitive endonucleases, BstN 1 and EcoRll. For construction of mitochondrial genomic library of ginseng, mtDNA was partially digested with EcoRl, and packaged with EMBL4 phage vector Genomic library was screened and purified for further research including restricttion mapping of ginseng mtDNA, and cloning of the genes. The gene of ATP synthase A subunit was cloned koto the purified EMBL4 library clone No. 16. Now, clone No. 16 is subcloned for structure gene sequence analysis.

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Cloning of the MLS Antibiotics Inducible Resistance Gene and Its Control Mechanism -Inducible Resistance to MLS Antibiotics of pMB4 Plasmid Isolated from Streptococcus sp. TR-1- (MLS계 항생물질 유도 내성 유전자의 크로닝과 유전자의 조절기전 -Streptococcus sp. TR-1에서 분리한 pMB 4 Plasmid의 MLS계 항생물질 유도내성-)

  • Jeong, Soon-Hak;Kwak, Jin-Hwan;Kim, Hee-Sun;Shim, Mi-Ja;Choi, Eung-Chil;Kim, Byong-Kak
    • YAKHAK HOEJI
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    • v.34 no.2
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    • pp.139-146
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    • 1990
  • Streptococcus sp. TR-1 which has inducible resistance to MLS antibiotics was isolated from soil samples in Korea. Streptococcus sp. TR-1 was cultured in Lysis broth, then a plasmid was isolated by modified Elliker method. Bacillus subtilis UOTO277 was transformed with that plasmid. This result showed that the plasmid has the gene relating with inducible resistance to MLS antibiotics. It was named pMB4 and its size was determined about 2.4 Kb by results of digestion with various restriction enzymes. Restriction endonuclease cleavage site map of pMB4 plasmid was made by double digestion of the plasmid. pMB4 plasmid has different restriction endonuclease site map from the other plasmids that have been discovered in Streptococcus sp. so far. And it could be identified that pMB4 plasmid does not have homology with ermK of Bacillus licheniformis EMR but has homology with ermC of Staphylococcus aureus from the results of Southern hybridization.

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A Profile of Naturally Occurring Plasmids from Selected Strains of Vibrios

  • Younghee Kim
    • Journal of Environmental Science International
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    • v.1 no.2
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    • pp.93.2-97
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    • 1992
  • The naturally occurring plasmids of Vibrio species have been isolated in part to investigate their genetic traits. Among six different Vibrio species tested, Vibrio anguillarum, Vibrio fluvialis, Vibrio vulnificus, Vibrio mimicus and Vibrio furnissi did not show any presence of plasmid. One environmental isolate of Vibrio pnrahemolyticus harboring plasmid was observed. The isolated plasmid was 8.7 kb by analysis with restriction endonuclease digestion. No common feature was shown relationships between the presence of plasmid and resistance against commonly used antibiotic compounds from the tested Vibrios. Key words . plasmid, Vibrio anguillarum, Vibrio fluvialis, Vibrio uulnincus, Vibrio mimicus, Vibrio furnissi, Vibrio parahemolyticus.

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A Micromethod for Rapid and Simple Isolation of Genomic DNA from Small Scale Culture of Bifidobacterium (소량의 Bifidobacterium 배양액에서 genomic DNA 추출을 위한 신속/간단한 방법)

  • Jeakal, Soo;Park, Hee-Kyung;Song, Ji-Eun;Heo, Tae-Ryeon;So, Jae-Seong
    • Microbiology and Biotechnology Letters
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    • v.23 no.6
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    • pp.781-783
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    • 1995
  • A method is described for the rapid and simple isolation of genomic DNA from 3 ml culture of Bifidobacterium. The method is expected to be used in gene manipulation of Bifidobacterium spp. The isolated DNA using this method is shown to be an excellent substrate for restriction endonuclease digestion and ligation with T4 DNA ligase.

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A Profile of Naturally Occurring Plasmids from Selected Strains of Vibrios

  • Kim, Young-Hee
    • Environmental Sciences Bulletin of The Korean Environmental Sciences Society
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    • v.1 no.2
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    • pp.93-97
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    • 1997
  • The naturally occurring plasmids of Vibrio species have been isolated in part to investigate their genetic traits. Among six different Vibrio species tested, Vibrio anguillarum, Vibrio fluvialis, Vibrio vulnficus, Vibrio mimicus and Vibrio furnissi did not show any presence of plasmid. One environmental isolate of Vibrio parahaemolyticus harboring plasmid was observed. The isolated plasmid was 8.7 kb by analysis with restriction endonuclease digestion. No common feature was shown relationships between the presence of plasmid and resistance against commonly used antibiotic compounds from the tested Vibrios.

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