• Title/Summary/Keyword: Phage

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Isolation and Characterization of the Streptococcus mutans from Korean Children with Caries

  • Kang, Kyung-Hee;Kim, Ji-Young;Nam, Jin-Sik;Jin, Ingn-Yol
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.341-347
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    • 2007
  • Streptococcus mutans, one of a major causal agents of dental caries, is component of the dental plaque and produces various organic acids such as lactic acid as the end-product of glycolysis. In this study, we isolated S. mutans from Korean children with caries and also investigated the expression of protein under acid stress. S. mutans was identified at the species level using a 16S ribosomal DNA sequencing comparison method. The primer specificity was tested on eleven S. mutans strains isolated from Korean children with caries. The data showed that eleven strains are S. mutans. At treatment of concentration of 20 mM lactic acid in the mid-log phage, K-7 exhibited the highest maximum culture OD compared with those of other groups. As a consequence, we examined the expression of protein under 20 mM lactic acid stress using S. mutans K-7. The results of 2D gel electrophoresis by image analysis showed that thirteen proteins are up-regulated under the stress. Further study is being focused on amino acid analysis by mass spectrometry in order to analyze those spots.

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ILLEGITIMATE RECOMBINATION MEDIATED BY MAMMALIAN DNA TOPOISOMERASE II

  • BAE YOUNG-SEUK;IKEDA HIDEO
    • Proceedings of the Microbiological Society of Korea Conference
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    • 1991.04a
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    • pp.82-96
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    • 1991
  • To understand the mechanism of illegitimate recombination in mammalian cells, we have examined the recombination role of DNA topoisomerase II (Topo II ). We found that purified calf thymus Topo II mediates recombination between two phage $\lambda$ DNA molecules in an in vitro system. The enzyme mainly produced a linear monomer recombinant DNA that can be packaged in vitro. Novobiocin and anti-calf thymus Topo II antibody inhibit this ATP-dependent recombination. The recombinant molecules contain duplications or deletion, and most crossovers take place between nonhomologous sequences of $\lambda$ DNA, as judged by the sequences of recombination junctions. In order to study the effects of Topo II on illegitimate recombination in mammalian cells, we have developed a new shuttle vector, pNKl, which contains three bacterial genes, amp(APR), galK and neo($Km^R$). Using this system, we have shown that a Topo II inhibitor, VM26, stimulated deletion formation in pNK1 DNA in monkey COS1 cells. Both in vitro and in vivo results suggest that Topo II participates in illegitimate recombination in mammalian cells.

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Cloning of Reverse Transcriptase Gene of Avian Sarcoma Virus (역전사효소(逆轉寫酵素) 유전자(遺傳子)의 cloning 에 관(關)한 연구(硏究))

  • Kim, Yong-Woong;Kim, Kwang-Sik;Suh, Yong-Tack;Guntaka, R.V.
    • Applied Biological Chemistry
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    • v.31 no.3
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    • pp.219-225
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    • 1988
  • Reverse transcriptase gene of Avian sarcoma virus(ASV) was cloned with a thermoinducible expression vector, pPL-lambda. E. coli N4830 which carries temperature sensitive cI857 la mbda repressor, was transformed with this pPL-pol plasmid DNA. The RNA transcribed by those tranoformants was isolated and analyzed. It was shown that the inserted reverse transcriptase gene of ASV was transcribed at high-level when cells were grown at high temperature.

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Pathogenicity of Salmonella typhimurium and S enteritidis isolated from domestic animals in Gyeongbuk province (경북지역 가축에서 분리된 Salmonella typhimurium과 S enteritidis의 병원성 시험)

  • Kim, Sang-Yun;Lee, Hee-Moo;Kim, Sin;Hong, Hyon-Pyo;Kwon, Heon-Il
    • Korean Journal of Veterinary Service
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    • v.24 no.1
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    • pp.69-82
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    • 2001
  • The result of studying the pathogenicity of Salmonella typhimurium and S enteritidis isolated from domestic animals in Gyeongbuk province were summarized as follows. In Congo-red binding test, S typhimurium had much more rough types than S enteritidis. In colicin production test, 4 strains of S typhimurium were positive but all of S enteritidis were negative. In hemolysin production test, all of S typhimurium and S enteritidis were negative. In Guinea pig serum resistant test, all of S typhimurium and S enteritidis were positive. As a result of pathogenicity test to mice, 54.4% of mice were died. Therefore, S typhimurium and S enteritidis were considered as highly pathogenic. S typhimurium DT104 and S enteritidis PT4 were more pathogenic to mice than other phage types of same serovar. S typhimurium and S enteritidis were considered not so pathogenic for 6-day-old chickens. The recovery rates of Salmonella stains from mice and chickens inoculated were 96.8%, and 54%, respectively. In chickens, proportional to the time from 2 weeks after challenge inoculation, the recovery rates were noticeably decreased.

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Design of Lactic Acid Bacteria Aiming at Probiotic Culture and Molecular Typing for Phyogenetic Identification (Probiotics용 유산균의 Design과 Molecular Typing에 의한 동정법)

  • Yoon, Sung-Sik
    • Journal of Dairy Science and Biotechnology
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    • v.18 no.1
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    • pp.47-60
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    • 2000
  • Over decades of work, the probiotic research has grown rapidly with a number of new cultures, which is claimed a variety of benefit. However, many of the specific effects attributed to the ingestion of probiotics remain convoluted and scientifically unsubstantiated. Accordingly, the scientific community faces a greater challenge and must objectively seek cause and effect relationships for many potential and currently investigated probiotic species. Rational selection and design of probiotics remains an important challenge and will require a solid information about the physiology and genetics of candidate strains relevant to their intestinal roles, functional activities, and interaction of with other resident micro flora. As far as beneficial culture of lactic acid bacteria (LAB) is concerned, simple, cost-effective, and exact identification of candidate strains is of foremost importance among others. Until recently, the relatedness of bacterial isolates has been determined sorely by testing for one or several phenotyphic markers, using methods such as serotyping, phage-typing, biotyping, and so forth. However, there are problems in the use of many of these phenotype-based methods. In contrast, some of newer molecular typing methods involving the analysis of DNA offer many advantages over traditional techniques. These DNA-based methods have the greater discriminatory power than that of phenotypic procedures. This review focuses on the importance and the basis of molecular typing methods along with some considerations on de-sign and selection of probiotic culture for human consumption.

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Studies on the hemolysin produced by Vibrio Vulnificus ys-1 (Vibrio vulnificus ys-1이 생산하는 hemolysin에 관한 연구)

  • 오양호;차미선;김민정
    • Journal of Life Science
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    • v.8 no.2
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    • pp.145-157
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    • 1998
  • We isolated 100 Vobrio sp. from marine products and sea from July to September, 1997. We attemped on purification of hemolysin produced by Vibrio sp. The growth, hemolysin production patterns by the 100 strains of Vibrio sp. showed identical, in general. V. unlnificus ys-1 produced hemolysis as the higtest titer. The optimal culture conditions for the hemolysin production by the V. vunificus ys-1 are followings; 1. Hemolysin production was optimal dering the late exponetial phage. 2. Maximal growth, hemolysin production were in heart infusion broth. 3. Maximal yields of hemolysin was obtained when the heart infusion broth had an intial pH of 8.0, 3$0^{\circ}C$, 3% NaCL. Hemolysin was purified from culture filtrate of the strain by ammonium sulfate recipitation, ion exchange and hydrophobic interaction chromatography. The results were as follows; 1. Hemogeneity of the purified hemolysin was demonstrated by revealing single band on SDS-PAGE. The molecular weight of purified hemolysin was 45KDa. 2. The absorbance rattern in ultraviolet wsa typical of those seen with most proteinb with 280nm. 3. Purified hemolysin was atable at 5$0^{\circ}C$ but 7$0^{\circ}C$ of the acivity was lost by heating for 30 min at 6$0^{\circ}C$/ Optimal temperature of purified hemolysin was 35$^{\circ}C$. 4. Purified hemolysin was stable at the pH range of 6~9, but in the less the pH5.0. above the pH 9.0, the hemolysin activity was lost completely.

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Biologically-Inspired Selective and Sensitive Trinitrotoluene Sensors Using Conjugated Lipid-like Polymer Nanocoatings for CNT-FET Sensors

  • Jaworski, Justyn;Kim, Tae-Hyun;Yokoyama, Keisuke;Chung, Woo-Jae;Wang, Eddie;Lee, Byung-Yang;Hong, Seung-Hun;Majumdar, Arun;Lee, Seung-Wuk;Kwon, Ki-Young
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.02a
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    • pp.495-495
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    • 2011
  • Miniaturized sensors capable of both sensitive and selective real-time monitoring of target analytes are tremendously valuable for various applications ranging from hazard detection to medical diagnostics. The wide-spread use of such sensors is currently limited due to insufficient selectivity for target molecules. We developed selective nanocoatings by combining trinitrotoluene (TNT) receptors bound to conjugated polydiacetylene (PDA) with single-walled carbon nanotube-field effect transistors (SWNT-FET). Selective binding events between TNT molecules and phage display derived TNT receptors were effectively transduced to sensitive SWNT-FET conductance sensors through the PDA coating. The resulting sensors exhibited unprecedented 1 fM sensitivity toward TNT in real time, with excellent selectivity over various similar aromatic compounds. Our biomimetic receptor coating approach may be useful for the development of sensitive and selective micro and nanoelectronic sensor devices for various other target analytes.

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Web-Based Computational System for Protein-Protein Interaction Inference

  • Kim, Ki-Bong
    • Journal of Information Processing Systems
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    • v.8 no.3
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    • pp.459-470
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    • 2012
  • Recently, high-throughput technologies such as the two-hybrid system, protein chip, Mass Spectrometry, and the phage display have furnished a lot of data on protein-protein interactions (PPIs), but the data has not been accurate so far and the quantity has also been limited. In this respect, computational techniques for the prediction and validation of PPIs have been developed. However, existing computational methods do not take into account the fact that a PPI is actually originated from the interactions of domains that each protein contains. So, in this work, the information on domain modules of individual proteins has been employed in order to find out the protein interaction relationship. The system developed here, WASPI (Web-based Assistant System for Protein-protein interaction Inference), has been implemented to provide many functional insights into the protein interactions and their domains. To achieve those objectives, several preprocessing steps have been taken. First, the domain module information of interacting proteins was extracted by taking advantage of the InterPro database, which includes protein families, domains, and functional sites. The InterProScan program was used in this preprocess. Second, the homology comparison with the GO (Gene Ontology) and COG (Clusters of Orthologous Groups) with an E-value of $10^{-5}$, $10^{-3}$ respectively, was employed to obtain the information on the function and annotation of each interacting protein of a secondary PPI database in the WASPI. The BLAST program was utilized for the homology comparison.

Characterization of Site-Specific Recombination by the Integrase MJ1 from Enterococcal Bacteriophage ${\Phi}FC1$

  • Park, Mi-Ok;Lim, Ki-Hong;Kim, Tae-Hyung;Chang, Hyo-Ihl
    • Journal of Microbiology and Biotechnology
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    • v.17 no.2
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    • pp.342-347
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    • 2007
  • Bacteriophage ${\Phi}FC1$ integrase (MJ1) was previously shown to perform a site-specific recombination between a phage attachment site (attP) and a host attachment site (attB) in its host, Enterococcus faecalis, and also in a non-host bacterium, Escherichia coli. Here, we investigated biochemical features of MJ1 integrase. First, MJ1 integrase could perform in vitro recombination between attP and attB in the absence of additional factors. Second, MJ1 integrase interacted with att sites. Electrophoretic mobility shift assays and DNase I footprinting revealed that MJ1 integrase could efficiently bind to all the att sites and that MJ1 integrase recognized relatively short sequences (${\sim}50bp$) containing an overlapping region within attB and attP. These results demonstrate that MJ1 integrase indeed catalyzes an integrative recombination between attP and attB, the mechanism of which might be simple and unidirectional, as found in serine integrases.

Draft genome sequence of Lactobacillus salivarius KLW001 isolated from a weaning piglet (이유자돈으로부터 분리한 Lactobacillus salivarius KLW001의 유전체 분석)

  • Jin, Gwi-Deuk;Lee, Jun-Yeong;Kim, Eun Bae
    • Korean Journal of Microbiology
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    • v.53 no.2
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    • pp.134-136
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    • 2017
  • Lactobacillus salivarius KLW001, a species of lactic acid bacteria (LAB), was isolated from a weaning piglet in a swine farm, South Korea, to develop an antimicrobial probiotic strain for piglets. Herein, we report the draft genome sequence of the strain. The genome contains 2,326,706 bp with a G+C content of 33.0% in 166 contigs (${\geq}500bp$). From the genome, we found out 4 genes related to antibiotic resistance, 36 genes for phages, 3 genes for bile hydrolysis, and 27 CRISPR spacers.