• Title/Summary/Keyword: DNase

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The Effect of Ethanol Extracts from Fish Flour on the Nucleic Acid Metabolism in Rats (소어분(小魚粉)의 주정추출성분(酒精抽出成分)이 백서(白鼠)의 핵산대사(核酸代謝)에 미치는 영향)

  • Oh, Seoung-Ho;Koh, Jin-Bog;Choi, Jeun-Duo;Lee, Myoung-Hoon
    • Journal of Nutrition and Health
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    • v.5 no.3
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    • pp.127-133
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    • 1972
  • This study was designed to observe the effect of ethanol extracts from fish flour on the nucleic acid metabolism in rats. Young rats, weighing 75-85g were used as the experimental animals and diet used were 8 kinds; diet supplemented with 10% fish flour, diets which were supplemented with the extracts and or remainders of fish flour after extracting by either 76% or 96% ethanol to the rice diet, respectively, and diet supplemented with 6% casein. After feeding corresponding diet for 40 days, RNA and DNA contents, and DNase activities in the liver, kidney and braid were determined. The results obtaioed from this study are summarized as follows: 1. The RNA contents of the ethanol-treatment groups are, in the liver and kidney, similar to, and in the brain, generally higher than, that of the control group. 2. The DNA contents of each organ show no difference between ethanol-treatment groups and control group, but in the liver, of ethanol extrat groups are lower than casein group. 3. the DNase activity of each organ in the ethanol-treatmeut groups, is generally lower than the control group. The above results indicate that ethanol extracts from fish flour have influence on the nucleic acid metabolism.

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Molecular Approaches to Determine the Character of Serratia marcescens Associated with the Insect Pathogenicity to Brown Planthopper (Serratia marcescens의 곤충 병원성 관련형질 탐색을 위한 분자생물학적 연구)

  • 김희규;배동원;박진희;윤한대
    • Korean journal of applied entomology
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    • v.32 no.3
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    • pp.330-337
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    • 1993
  • A bacterium, pathogenic to Nilaparvata lugens Stal. causing high mortality in 3~5 days, were selected and identified as Serratuz marcescens biotype A2a which is not a nosocomlally infective strain. In order to determine the characters of Serratia marcesce'1lS associated with insect pathogenicity, Tn5 mutagenesis was carried out by conjugating with E. coli pJB4J1. Transconjugants were plate-assayed for missing chitinase, protease and DNase activity. A protease negative mutant was selected for missing JOseet pathogenicity. SEM and TEM revealed the presence of bacterial cells in the epithelial tissue of inner abdomal tissue of the hypodermic layer of abdomen. Such a colonization was limmited to the subjacent tissue inside the intacL cuticular epidermis. These observation supported our result of pathogenicity tests of transconjugants.

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Complexation of Adiponectin-encoding Plasmid DNA with Rosiglitazone-loaded Cationic Liposomes

  • Davaa, Enkhzaya;Jeong, Ui-Hyeon;Shin, Baek-Ki;Choi, Soon-Gil;Myung, Chang-Seon;Park, Jeong-Sook
    • Journal of Pharmaceutical Investigation
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    • v.40 no.6
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    • pp.357-362
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    • 2010
  • To enhance therapeutic effects of insulin-sensitizing adipokine, ADN gene and potent agonists, rosiglitazone for the $PPAR{\gamma}$, cationic liposomes as non-viral vectors were formulated. The particle size and zeta potential of drug loaded and unloaded cationic liposomes were investigated. The complex formation between cationic liposomes and negatively charged plasmid DNA was confirmed and the protection from DNase was observed. In vitro transfection was investigated in HepG2, HeLa, and HEK293 cells by mRNA expression of ADN. Encapsulation efficacy of rosiglitazone-loaded liposomes was determined by UV detection. Particle sizes of cationic liposomes were in the range of 110-170 nm and those of rosiglitazone-loaded cationic liposomes were in the range of 130-180 nm, respectively. Gel retardation of complexes indicated that the complex was formed at weight ratios of cationic lipid to plasmid DNA higher than 20:1. Both complexes protected plasmid DNA from DNase either drug free or drug loading. Encapsulation efficiency of rosiglitazone-loaded emulsion was increased by drug dose. The mRNA expression levels of ADN were dose-dependently increased in cells transfected with plasmid DNA. Therefore, cationic liposomes could be potential co-delivery system for drug and gene.

Understanding Epistatic Interactions between Genes Targeted by Non-coding Regulatory Elements in Complex Diseases

  • Sung, Min Kyung;Bang, Hyoeun;Choi, Jung Kyoon
    • Genomics & Informatics
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    • v.12 no.4
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    • pp.181-186
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    • 2014
  • Genome-wide association studies have proven the highly polygenic architecture of complex diseases or traits; therefore, single-locus-based methods are usually unable to detect all involved loci, especially when individual loci exert small effects. Moreover, the majority of associated single-nucleotide polymorphisms resides in non-coding regions, making it difficult to understand their phenotypic contribution. In this work, we studied epistatic interactions associated with three common diseases using Korea Association Resource (KARE) data: type 2 diabetes mellitus (DM), hypertension (HT), and coronary artery disease (CAD). We showed that epistatic single-nucleotide polymorphisms (SNPs) were enriched in enhancers, as well as in DNase I footprints (the Encyclopedia of DNA Elements [ENCODE] Project Consortium 2012), which suggested that the disruption of the regulatory regions where transcription factors bind may be involved in the disease mechanism. Accordingly, to identify the genes affected by the SNPs, we employed whole-genome multiple-cell-type enhancer data which discovered using DNase I profiles and Cap Analysis Gene Expression (CAGE). Assigned genes were significantly enriched in known disease associated gene sets, which were explored based on the literature, suggesting that this approach is useful for detecting relevant affected genes. In our knowledge-based epistatic network, the three diseases share many associated genes and are also closely related with each other through many epistatic interactions. These findings elucidate the genetic basis of the close relationship between DM, HT, and CAD.

Development of Reverse Transcriptase-Polymerase Chain Reaction of fimA Gene to Detect Viable Salmonella in Milk (우유 내 활력있는 Salmonella를 검출하기 위한 fimA 유전자의 역전사중합효소 연쇄반응의 개발)

  • Choi, S.H.;Lee, S.B.
    • Journal of Animal Science and Technology
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    • v.46 no.5
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    • pp.841-848
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    • 2004
  • Rapid detection of viable Salmonella in pasteurized milk is important to protect public health from food poisoning. Reverse transcriptase-polymerase chain reaction(RT-PCR) is recognized as a molecular genetical method to differentiate between live and dead bacteria The RT-PCR in this study was designed to detect specifically viable Salmonella in milk by using the primers whose nucleotide sequences were determined based on fimA gene which encodes the submit of type 1 fimbriae. Treatment of RNA preparation with RNase-free DNase was adequate enough to destroy DNA, which may otherwise be amplified in the RT PCR Seven strains of Salmonella were detected in the RT-PCR but Escherichia coli, Shigella sonnei, Citrobacter freundii, and Klebsiella pneumoniae were not. $10^7/ml$ and $10^6/ml$ of dead Salmonella which were heat-treated in milk were detectable by using the RT-PCR but $10^5{\sim}10/ml$ of the dead bacteria were not. The sensitivity of the RT-PCR in detecting viable Salmonella was 100 cells/ml.

Differential Regulation of the Caprine ${\beta}$-Lactoglobulin Gene Promoter in the Cultured Mammary HC11 Cells

  • Kim, Jae-Man
    • Animal cells and systems
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    • v.1 no.2
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    • pp.345-350
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    • 1997
  • The ${\beta}$-Lactoglobulin (BLG) gene expression is differentially regulated during development of the mammary tissues. Such differential regulation of the BLG gene expression can be reiterated in the cultured mammary HC11 cells. In the growing non-confluent HC11 cells, the BLG promoter activity was shown to be partially repressed by the upstream regulatory sequence. The repression was gradually diminished and switched to activation as the cells grew confluent. The differential regulation of the BLG promoter was controlled by the 5'-regulatory sequence located at the upstream of 205 bp. Electromobility shift assay showed that nuclear extract from HC11 cells differentially bound on the regulatory sequence, depending on the cell confluency, which was in accordance with the differential transcriptional activity. DNase I foot-print assay, however, revealed that all nuclear extracts presented the same foot-prints, regardless of confluency of HC11 cells. These results suggest that differential regulation BLG gene expression by the 5'-regulatory sequence may be accomplished by competitive and/or cooperative binding of differential regulatory factors on the same regulatory element.

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Bacteriocins: Assay, Biochemistry, and Mode of Action

  • Paik, Hyun-Dong
    • Preventive Nutrition and Food Science
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    • v.1 no.2
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    • pp.269-277
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    • 1996
  • Bacteriocins are proteins produced by a heterogeneous group of bacteria that have a bactericidal effect on closely related organisms. Recently, bacteriocins from lactic acid bacteria and other food-related organisms have been the subject of much research because of their potential as food biopreservatives. Various modifications of agar plate diffusion assays are the most widely used methods even though the limitations of such assays are generally recognized. The ability to obtain a concentrated crude preparation on bacteriocin by optimizing production parameters greatly simplifies recovery of bacteriocin on subsequent purification steps. Some studies performed to optimize bacteriocins have been purified to homogeneity, and the amino acid sequences of many of these purified bacteriocins have been determined. Obtaining characterization data on purified bacteriocin will minimize the risk of overlapping of research and confusion on identification of these compounds. Several me-chanisms leading to cell death have been hypothesized. These include depletion of the proton motive force(PMF) across the cell membrane: RNase and/or DNase activity within the sensitive cell; and pore formation and lysis of sensitive cells at the cell membrane.

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Characterization of Linear Polymer-Dendrimer Block Copolymer/Plasmid DNA Complexes: Formation of Core-shell Type Nanoparticles with DNA and Application to Gene Delivery in Vitro

  • Choi, Joon-Sig;Choi, Young-Hun;Park, Jong-Sang
    • Bulletin of the Korean Chemical Society
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    • v.25 no.7
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    • pp.1025-1030
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    • 2004
  • A hybrid linear polymer-dendrimer block copolymer, poly(ethylene glycol)-block-poly(L-lysine) dendrimer, was synthesized and introduced to form polyionic complexes with DNA. The copolymer formed core-shell type nanoparticles with plasmid DNA. From dynamic light scattering experiments, the mean diameter of the polyplexes was observed to be 154.4 nm. The complex showed much increased water solubility compared to poly(L-lysine). The plasmid DNA in polyplexes was efficiently protected from the enzymatic digestion of DNase I. The cytotoxicity and transfection efficiency for 293 cells was measured in comparison with poly(Llysine).

Identification of the Fur-Binding Site in Regulatory Region of the Vulnibactin-Receptor Gene in Vibrio vulnificus

  • Lee, Hyun-Jung;Lee, Kyu-Ho
    • Journal of Microbiology and Biotechnology
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    • v.22 no.1
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    • pp.46-49
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    • 2012
  • The Vibrio vulnificus vuuA gene, of which expression is repressed by a complex of iron and ferric uptake regulator (Fur), was characterized to localize the Fur-binding site in its upstream regulatory region. In silico analysis suggested the presence of two possible Fur-binding sites; one is a classical Fur-box and the other is a previously reported distinct Fur-binding site. Site-directed mutagenesis and DNase I protection assays revealed the binding site for the iron-Fur complex, which includes an extended inverted repeat containing a homologous sequence to the classical Fur-box.

Restriction pattern of the nucleic acid of Synechococcus sp. cyanophage

  • Park, Jong-Geun;Kim, Min;Choi, Yong-Keel;Yoon, Sung-Nyo
    • Journal of Microbiology
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    • v.34 no.1
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    • pp.1-6
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    • 1996
  • The nucleic acid of Synechococcus sp. cyanophage was identified as double-stranded DNA by the result of digestion with enzymes such as exonucleases, DNase, and S1 nuclease, and by acridine orange staining. The cyanophage DNA was cleaved with several restriciton ehdonucleases such as ApaI, BamHI, Bg/II, HaeIII, Eco RI, HindIII, PstI, AND aPAI gave the clearest sets of bands on agarose gels and the fragment numbers for each were 12, 20, 29, 20, and 7, respectively. The sums of the size from Bam HI and PstI digestions were estimated approximately 227$\pm$4 kb, which are in agreement with the result of the pulsed field gel electrphoresis. This virus is thought to have the largest genosome among those of known cyanophages, which corresponds to the largest haed ot 90 nm when compared with the head sizes of cyanophages discovered since 1963.

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