• Title/Summary/Keyword: Phage

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Cyclic Peptides as Therapeutic Agents and Biochemical Tools

  • Joo, Sang-Hoon
    • Biomolecules & Therapeutics
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    • v.20 no.1
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    • pp.19-26
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    • 2012
  • There are many cyclic peptides with diverse biological activities, such as antibacterial activity, immunosuppressive activity, and anti-tumor activity, and so on. Encouraged by natural cyclic peptides with biological activity, efforts have been made to develop cyclic peptides with both genetic and synthetic methods. The genetic methods include phage display, intein-based cyclic peptides, and mRNA display. The synthetic methods involve individual synthesis, parallel synthesis, as well as split-and-pool synthesis. Recent development of cyclic peptide library based on split-and-pool synthesis allows on-bead screening, in-solution screening, and microarray screening of cyclic peptides for biological activity. Cyclic peptides will be useful as receptor agonist/antagonist, RNA binding molecule, enzyme inhibitor and so on, and more cyclic peptides will emerge as therapeutic agents and biochemical tools.

Fast Genetic Variation among Coliphage Quasispecies Revealed by a Random Amplified Polymorphic DNA (RAPD) Analysis

  • Kwon, Oh-Sik;Lee, Jae-Yung
    • Journal of Microbiology
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    • v.34 no.2
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    • pp.166-171
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    • 1996
  • Genetic analysis was conducted on newly isolated coliphages form soil by using a RAPD assay. From the initial result, the coliphages were turned out to be different form one another but were closely related to .psi..lambda. due to the fact that they shared the samed RAPD maker in which other T phage testings failed to show. By using the primers EC01 or EC02, a fast genetic mutation of .psi.C1 was found by producing specific RAPD markers on the phages from the first filial progeny to the second filial progeny. When we made a RAPD assay with combined primers (EC01, EC05 and EC08), the genetic mutation was again confirmed in .psi.C1. The assay detection showed mutations in other coliphages such as .psi.C2 and .psi.C3 by revealing specific RAPD bands among different progeny phages, where genetic instability of the coliphages in implied.

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Application of LFH-PCR for the Disruption of SpoIIIE and SpoIIIG of B. subtilis

  • Kim, June-Hyung;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.5 no.5
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    • pp.327-331
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    • 2000
  • The application of LFH-PCR(long flanking homology region-PCR) for Bacillus subtilis gene disruption is presented. Without plasmid- or phage-vector construction, only by PCR, based on a DNA sequence retrieved from B. subtilis genome data base, kanamycin resistance gene was inserted into two genes of B. subtilis involved in sporulation, spoIIIE and spoIIIG. The effect of gene disruption on subtilisin expression was examined and the sporulation frequency of two mutants was compared to that of the host strain. For this purpose, only 2 or 3 rounds of PCR were required with 4 primers. We first demonstrated the possibility of LFH-PCR for rapid gene disruption to characterize an unknown functional gene of B. subtilis or other prokaryote in the genomic era.

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The Studies of Copolymer, Poly (3-hydroxybutyrate-co-3-hydroxy valerate) Synthesis in Recombinant pha+ Escherichia coli

  • Lee, Ha-Young;Rhie, Ho-Gun
    • Proceedings of the Zoological Society Korea Conference
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    • 1996.10a
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    • pp.195.2-195
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    • 1996
  • acs gene cloning was constructed by subcloning the 2.2-kb MunI-MunI restriction fragment of 638 and 639 which include acs gene from the kohara phage into the unique EcoRI site of pUC18 and pJM9131 containing the PHA biosynthesis genes. Then recombinant E. coli fadRatoC(Con) mutants containing the polyhydroxyalkanoate(PHA) biosynthesis genes are able to incoporate s significant levels of 3-hydroxyvalerate (3HV) into the copolymer [P(3HB-co-3HV)]. Quantitative determination of PHB and P(3HB-co-3HV) was performed by gas-chromatographic analysis of extracts obtained from methanolysis of lyophilized cells.

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Isolation of Proteinase Inhibitor II Genes from Potato (감자로부터 단백질분해효소 억제제 II 유전자의 분리)

  • 이종섭
    • Journal of Plant Biology
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    • v.32 no.2
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    • pp.79-87
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    • 1989
  • Southern hybridization of genomic DNAs with radioactively labeled cDNA of tomato proteinase inhibitor II revealed that proteinase inhibitor II proteins in potato plants are encoded by a family of about 10 related sequences. Screening of potato EcoRI genomic library with the cDNA resulted in isolation of 13 recombinant phage clones which carry 3 different genomic regions. Of these clones, clones 8, 18, and 39 were subjected to restriction mapping and subcloning. Further characterization of the subclones of clones 8, 18 and 39 indicated that two inhibitor II genes are present on a 8.0 kb EcoRI fragment of clone 8, one on 3.3 and 0.8 kb EcoRI fragments of clone 18 and two genes on a 13.5 kb EcoRI fragment of clone 39.

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Inhibition of SV40 DNA replication in simian cell by bacteriophage M13 DNA sequences (원숭이 신장 세포에서 M13 DNA에 의한 SV40 DNA 복제 억제 현상에 대하여)

  • 김연수;구용의;강현삼
    • Korean Journal of Microbiology
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    • v.26 no.3
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    • pp.162-166
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    • 1988
  • Bacteriophage M13 DNAs carrying the wild type or base substituted SV40 DNA replication origins were used for replication assay. In vivo and in vitro assay with African green monkey cell line COS-1 showed that the replication of M13-SV40 recombinant DNAs was restricted like a pBR322 SV40 recombinant DNA(Lusky and Botchan, 1981). Furthermore, recombinant phage DNAs isolated from the transfected siminan cells subsequently show a reduced ability to retransform E. coli. But pATSV-W(Kim et al., 1988) was replicated in COS-1 cells normally. We think that a poison sequence may exist on bacteriophage M13 DNA like pBR322.

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Selection of Peptides Binding to HCV E2 and Inhibiting Viral Infectivity

  • Hong, Hye-Won;Lee, Seong-Wook;Myung, Hee-Joon
    • Journal of Microbiology and Biotechnology
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    • v.20 no.12
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    • pp.1769-1771
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    • 2010
  • The envelope glycoprotein E2 of hepatitis C virus (HCV) binds to various cell surface receptors for viral infection. We performed biopanning against this protein and selected peptides from phage display peptide libraries. Two short peptides, pep7-1 and pep12-1, were selected and their ability to inhibit the infection process was investigated. When pep7-1 was present, the infectivity of HCV particles in cell culture was notably decreased. This decrease was demonstrated by Western blot analysis, immunofluorescence assay, and reverse transcription PCR assay. However, pep12-1 showed little inhibitory effect on HCV infection.

Selective overproduction of chloramphenicol acetyltransferase in the T7 expression system (T7 발현체계에서 chloramphenicol acetyltransferase의 선택적 과잉생산)

  • 김한복;강창원
    • Korean Journal of Microbiology
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    • v.27 no.4
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    • pp.317-322
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    • 1989
  • A gene can be selectively overexpressed in E. coli by utilizing the phage T7 RNA polymerase's stringent recognition and active transcription of the T7 promoter. The T7 expression system was constructed such that the T7 RNA polymerase gene is under the control of lacUV5 promoter in one plasmid, and that the target gene, the promoterless chloramphenicol acetyltransferase (CAT) gene with E. coli ribosome binding site is under the control of T7 promoter in the other plasmid. Only the E. coli cells containing both plasmids show high resistance to chloramphenicol. When the copy number of the runaway plasmid containing the polymerase gene was varied by a temperature shift, amounts of the CAT protein synthesized upon induction was correspondingly changed as shown in SDS gel electrophoresis.

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Phage Litmus: Biomimetic Virus-Based Colorimetric Sensors for Explosive Detection

  • O, Jin-U
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.90.1-90.1
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    • 2013
  • Nature utilizes various of the colorization process. Some species of birds can express their mood of tempers by changing their collagen structures on skin. For example, turkey can change their skin color by expansion of the collagen structures, which are associated with the distinct color changes. Here, we developed bioinspired virus-based colorimetric sensors which can be genetically tuned for target molecule. Using M 13 bacteriophage, we fabricated responsive self-assembled color matrices composed of quasi-ordered fiber bundle structures. These virus matrices can exhibit color change by stimuli through fiber bundle structure modulation. Upon exposure of volatile organic compounds, the resulting multi-colored matrices exhibited distinct color changes with different ratios that can be recognized by the naked eyes. Using the directed evolutionary approaches, we genetically engineered the virus matrix to incorporate binding motif for explosive detection (i.e., trinitrotoluene (TNT)). Through utilizing a common handheld device (i.e., iPhone), we could distinguish TNT molecules down to 20 ppb in a selective manner. Our novel biomimetic virus colorimetric sensor can overcome current limitation for low response selectivity.

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Isolation of Bacteriophage from Haloarcular sp, EH-1 (Haloarcular sp. EH-1에 의한 bacteriophage의 분리)

  • 정명주
    • Journal of Life Science
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    • v.13 no.4
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    • pp.505-510
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    • 2003
  • The extremely halophilic archaebacteriurn Haloarcular sp. EH-1 was isolated from solar salts. Halophages found in Haloarcular sp. EH-1 were isolated from fermented anchovy sauce. Halophages were isolated from fermented anchovy sauce using Haloarcular sp. EH-1 as a host bacterium. The isolated halophage produced 0.5∼l.0 mm in diameter clear plaques. The halophage consists of an symmetrical head, measuring 68 nm in diameter, and a contractile tail, 100 nm long and base plates were observed. Total size of phage DNA genome obtained 20 Kbp and its sequence homology was 52.87% with H. Salinarium.