• 제목/요약/키워드: DNA interaction

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Isolation and Characterization of a Bacteriophage Preying an Antifungal Bacterium

  • Rahimi-Midani, Aryan;Kim, Kyoung-Ho;Lee, Seon-Woo;Jung, Sang Bong;Choi, Tae-Jin
    • The Plant Pathology Journal
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    • v.32 no.6
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    • pp.584-588
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    • 2016
  • Several Bacillus species were isolated from rice field soils, and 16S rRNA gene sequence analysis showed that Bacillus cereus was the most abundant. A strain named BC1 showed antifungal activity against Rhizoctonia solani. Bacteriophages infecting strain BC1 were isolated from the same soil sample. The isolated phage PK16 had an icosahedral head of $100{\pm}5nm$ and tail of $200{\pm}5nm$, indicating that it belonged to the family Myoviridae. Analysis of the complete linear dsDNA genome revealed a 158,127-bp genome with G + C content of 39.9% comprising 235 open reading frames as well as 19 tRNA genes (including 1 pseudogene). Blastp analysis showed that the proteins encoded by the PK16 genome had the closest hits to proteins of seven different bacteriophages. A neighbor-joining phylogenetic tree based on the major capsid protein showed a robust clustering of phage PK16 with phage JBP901 and BCP8-2 isolated from Korean fermented food.

Rice NAC proteins act as homodimers and heterodimers

  • Jeong, Jin Seo;Park, Yeong Taek;Jung, Harin;Park, Su-Hyun;Kim, Ju-Kon
    • Plant Biotechnology Reports
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    • v.3 no.2
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    • pp.127-134
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    • 2009
  • Members of the NAM-ATAF-CUC (NAC) protein family are plant-specific transcription factors that contain a highly conserved N-terminal NAC-domain and diverse C-terminal regions. They have been implicated in plant development and abiotic stress responses. To identify interacters of rice NAC-domain proteins (OsNACs), we performed yeast two-hybrid screening of rice cDNA library using OsNAC5 as a bait, and the results showed that OsNAC5 interacts with other OsNACs including itself. To delineate an interacting domain, a series of deletion constructs of four OsNACs were made and transformed into yeast in various combinations. The results revealed that the conserved NAC domain of OsNACs plays a primary role in homodimer and heterodimer formation, and a part of C-terminal sequence is also necessary for the interaction. In vitro pull-down assays using recombinant OsNAC proteins verified the dimer formations, together suggesting that OsNACs may act by forming homodimers and/or heterodimers in plants.

Characterization of γ-Aminobutyric acid(GABA) produced by a lactic acid bacterium from button mushroom bed

  • Lee, Yun-Seok;Song, Tae-Young;Kong, Won-Sik;Yoon, Min-Ho
    • Journal of Mushroom
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    • v.11 no.4
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    • pp.181-186
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    • 2013
  • ${\gamma}$-Aminobutyric acid(GABA) is a four carbon non-protein amino acid that has several well-known physiological functions, such as a postsynaptic inhibitory neurotransmitter in the brain and induction of hypotensive and tranquilizer effects. A lactic acid bacterium was isolated from button mushroom bed, which is showing high GABA productivity by TLC or HPLC analysis. The strain was identified as Lactobacillus hilgardii by analysis of 16S rDNA gene sequence. When the maximum production of GABA by L. hilgardii was investigated with various concentration of monosodium glutamate, the yield of GABA reached to be 53.65 mM at 1% mono sodium glutamate (MSG) in flask cultivation. A Glutamate decarboxylase (GAD) enzyme, which was known to convert MSG to GABA, was purified from a cell-free extract of L. hilgardii and the molecular weights of purified GAD was estimated to 60,000 by SDS-PAGE. The optimum pH and temperature of GAD were at pH4.6 and at $37^{\circ}C$, respectively. The GAD activity was increased by the addition of sulfate ions such as ammonium sulfate, sodium sulfate and magnesium sulfate, indicating that the increase of hydrophobic interaction causes the increase of GAD activity.

The Effect of Water in Four Adenine-Thymine and Three Guanine-Cytosine Pairs: Combining Quantum and Statistical Mechanics

  • Lee, Jinkeong;Ham, Sihyun
    • Proceeding of EDISON Challenge
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    • 2015.03a
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    • pp.151-155
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    • 2015
  • The molecular interactions between the nucleic acid bases and water molecules are important in organism. Despite Adenine-Thymine Hoogsteen base pair and Guanine-Cytosine Watson-Crick base pair have been demonstrated to be most stable in a gas phase, the effect of water on the stability of these base pairs remains elusive. Here we report the structural and thermodynamic characteristics on possible Adenine-Thymine and Guanine-Cytosine base pairs in a gas phase as well as in an aqueous phase by using quantum mechanical method and statistical mechanical calculations. First, we optimized the direct base-pair interaction energies of four Adenine-Thymine base pairs (Hoogsteen base pair, reverse Hoogsteen base pair, Watson-Crick base pair, and reverse Watson-Crick base pair) and three Guanine-Cytosine base pairs (GC1 base pair, GC2 base pair, and Watson Crick base pair) in a gas phase at the $B3LYP/6-31+G^{**}$ level. Then, the effect of solvent was quantified by the electronic reorganization energy and the solvation free energy by statistical mechanical calculations. Thereby, we discuss the effect of water on the stability of Adenine-Thymine and Guanine-Cytosine base pairs, and argue why Adenine-Thymine Watson-Crick base pair and Guanine-Cytosine Watson-Crick base pair are most stable in an aqueous environment.

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Antibacterial and Pharmacological Evaluation of Fluoroquinolones: A Chemoinformatics Approach

  • Sood, Damini;Kumar, Neeraj;Singh, Aarushi;Sakharkar, Meena Kishore;Tomar, Vartika;Chandra, Ramesh
    • Genomics & Informatics
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    • v.16 no.3
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    • pp.44-51
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    • 2018
  • Fluoroquinolone (FQ) antibiotics are an important class of synthetic antibacterial agents. These are the most extensively used drugs for treating bacterial infections in the field of both human and veterinary medicine. Herein, the antibacterial and pharmacological properties of four fluoroquinolones: lomefloxacin, norfloxacin, ciprofloxacin, and ofloxacin have been studied. The objective of this study was to analyze the antibacterial characteristics of the different fluoroquinolones. Also, the pharmacological properties of the compounds including the Lipinski rule of five, absorption, distribution, metabolism, and excretion, LD50, drug likeliness, and toxicity were evaluated. We found that among all four FQ molecules, ofloxacin showed the highest antibacterial activity through in silico assays with a strong interaction (-38.52 kJ/mol) with the antibacterial target protein (topoisomerase-II DNA gyrase enzyme). The pharmacological and pharmacokinetic analysis also showed that the compounds ciprofloxacin, ofloxacin, lomefloxacin and norfloxacin have good pharmacological properties. Notably, ofloxacin was found to possess an IGC50 (concentration needed to inhibit 50% growth) value of $0.286{\mu}g/L$ against the Tetrahymena pyriformis protozoa. It also tested negative for the Ames toxicity test, showing its non-carcinogenic character.

BETTER UNDERSTANDING OF THE BIOLOGICAL EFFECTS OF RADIATION BY MICROSCOPIC APPROACHES

  • Kim, Eun-Hee
    • Nuclear Engineering and Technology
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    • v.40 no.7
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    • pp.551-560
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    • 2008
  • Radiation has stochastic aspects in its generation, its choice of interaction mode during traveling in media, and its impact on living bodies. In certain circumstances, like in high dose environments resulting from low-LET radiation, the variance in its impact on a target volume is negligible. On the contrary, in low dose environments, especially when they are attributed to high-LET radiation, the impact on the target carries with it a large variance. This variation is more significant for smaller target volumes. Microdosimetric techniques, which have been developed to estimate the distribution of radiation energy deposited to cellular and subcellular-sized targets, contrast with macrodosimetric techniques which count only the average value. Since cells and DNA compounds are the critical targets in human bodies, microdosimetry, or dose estimation by microscopic approach, helps one better analyze the biological effects of radiation on the human body. By utilizing microbeam systems designed for individual cell irradiation, scientists have discovered that human cells exhibit radiosensitive reactions without being hit themselves (bystander effect). During the past 10 or more years, a new therapeutic protocol using discontinuous multiple micro-slit beams has been investigated for its clinical application. It has been suggested that the beneficial bystander effect is the essence of this protocol.

Expression and Purification of a Cathelicidin-Derived Antimicrobial Peptide, CRAMP

  • Park Eu-Jin;Chae Young-Kee;Lee Jee-Young;Lee Byoung-Jae;Kim Yang-Mee
    • Journal of Microbiology and Biotechnology
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    • v.16 no.9
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    • pp.1429-1433
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    • 2006
  • Application of recombinant protein production and particularly their isotopic enrichment has stimulated development of a range of novel multidimensional heteronuclear NMR techniques. Peptides in most cases are amenable to assignment and structure determination without the need for isotopic labeling. However, there are many cases where the availability of $^{15}N$ and/or $^{13}C$ labeled peptides is useful to study the structure of peptides with more than 30 residues and the interaction between peptides and membrane. CRAMP (Cathelicidin-Related AntiMicrobial Peptide) was identified from a cDNA clone derived from mouse femoral marrow cells as a member of cathelicidin-derived antimicrobial peptides. CRAMP was successfully expressed as a GST-fused form in E. coli and purified using affinity chromatography and reverse-phase chromatography. The yield of the CRAMP was 1.5 mg/l 1. According to CD spectra, CRAMP adopted ${\alpha}$-helical conformation in membrane-mimetic environments. Isotope labeling of CRAMP is expected to make it possible to study the structure and dynamic properties of CRAMP in various membrane systems.

Novel Trimeric Complex for Efficient Uptake of Plasmid Vector into HepG2 Cells

  • Joo, Jong-Hyuck;Park, Jong-Gu
    • Biomedical Science Letters
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    • v.9 no.2
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    • pp.67-74
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    • 2003
  • Viral and non-viral vectors have been used in the delivery of genetic materials into animal cells and tissues, with each approach having pros and cons. Non-viral vectors have many useful merits such as easy preparation, low immunity and size tolerance of a transgene when compared to those of viral vectors. Delivery specificity may be achieved by complex formation between receptor ligands and a non-viral vector. In the present study, non-viral vector systems are investigated in an effort to find a practical delivery means for gene therapy, Receptor-ligand interaction between transferrin-receptor and transferrin was utilized for efficient gene transfer into cancer cells. A plasmid vector, pcDNA3 (LacZ) was ligated with a small duplexed oligo fragment in which a Biotin- VN$^{TM}$ phosphoramidite was placed in the middle of the oligo. The plasmid vector labeled by biotin was then conjugated with biotin-labeled transferrin via streptavidin. This trimeric conjugates were delivered to a hepatoma cell line, HepG2. The delivery efficiency of the trimeric conjugate was 2-fold higher than that of cationic liposomes used for transfection of a plasmid vector. These results demonstrate that a plasmid vector can be efficiently transferred into cells by forming a trimeric complex of plasmid vector-linker-ligand.

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The Effect of Cobrotoxin on $NF-{\kappa}B$ binding Activity in Raw264.7 cells

  • Yoo, Jae-Ryong;Song, Ho-Sueb
    • Journal of Acupuncture Research
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    • v.22 no.2
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    • pp.133-139
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    • 2005
  • Cobrotoxin, a venom of Vipera lebetina turanica, is a group of basic peptidescomposed of 233 amino acids with six disulfide bonds formed by twelve cysteins. NF-kB is activated by subsequent release of inhibitory IkB and translocation of p50. Since sulfhydryl group is present in kinase domain of p50 subunit of NF-kB, cobrotoxin could modify NF-kB activity by protein-protein interaction. We therefore examined effect of cobrotoxin on NF-kB activities in lipopolysaccharide (LPS) and sodium nitroprusside (SNP)-stimulated Raw 264.7 mouse macrophages. Cobrotoxin suppressed the LPS and SNP-induced release of IkB and p50 translocation resulted in inhibition of DNA binding activity of NF-kB. Inhibition of NF-kB resulted in reduction of the LPS and SNP-induced production of inflammatory mediators NO and PGE2 generation. The inhibitory effect of cobrotoxin on the NF-kB activity were blocked by addition of reducing agents dithiothreitol and glutathione. These results demonstrate that cobrotoxin inhibits activation of NF-kB, and suggest that pico to nanomolar range of cobrotoxin could inhibit the expression of genes in the NF-kB signal pathway.

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hpvPDB: An Online Proteome Reserve for Human Papillomavirus

  • Kumar, Satish;Jena, Lingaraja;Daf, Sangeeta;Mohod, Kanchan;Goyal, Peyush;Varma, Ashok K.
    • Genomics & Informatics
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    • v.11 no.4
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    • pp.289-291
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    • 2013
  • Human papillomavirus (HPV) infection is the leading cause of cancer mortality among women worldwide. The molecular understanding of HPV proteins has significant connotation for understanding their intrusion in the host and designing novel protein vaccines and anti-viral agents, etc. Genomic, proteomic, structural, and disease-related information on HPV is available on the web; yet, with trivial annotations and more so, it is not well customized for data analysis, host-pathogen interaction, strain-disease association, drug designing, and sequence analysis, etc. We attempted to design an online reserve with comprehensive information on HPV for the end users desiring the same. The Human Papillomavirus Proteome Database (hpvPDB) domiciles proteomic and genomic information on 150 HPV strains sequenced to date. Simultaneous easy expandability and retrieval of the strain-specific data, with a provision for sequence analysis and exploration potential of predicted structures, and easy access for curation and annotation through a range of search options at one platform are a few of its important features. Affluent information in this reserve could be of help for researchers involved in structural virology, cancer research, drug discovery, and vaccine design.