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Phage Particles (phage + particle)
Selected AbstractsUltrastructural and electron energy-loss spectroscopic analysis of an extracellular filamentous matrix of an environmental bacterial isolateENVIRONMENTAL MICROBIOLOGY, Issue 9 2007Uta Böckelmann Summary Strain F8, a bacterial isolate from ,river snow', was found to produce extracellular fibres in the form of a filamentous network. These extracellular filaments, which were previously shown to be composed of DNA, have been studied for the first time by ultrastructural and electron energy-loss spectroscopy in the present work. ,Whole mount' preparations of strain F8 indicate these polymers are ultrastructurally homogeneous and form a network of elemental filaments, which have a width of 1.8,2.0 nm. When incubated at pH 3.5 with colloidal cationic ThO2 tracers they become intensely stained (electron dense), affording direct evidence that the fibres are negatively charged and thus acidic chemically. Elemental analysis of the extracellular filaments by Energy-filtered Transmission Electron Microscopy revealed phosphorus to be the main element present and, because pretreatment of F8 cells with DNase prevented thorium labelling, the fibres must be composed of extracellular DNA (eDNA). Neither ultrathin sections nor ,whole mount negative stain' caused DNA release by general cell lysis. Additionally, cells infected with phages were never observed in ultrathin sections and phage particles were never detected in whole mount samples, which rules out the possibility of phages being directly involved in eDNA release. [source] Phage display particles expressing tumor-specific antigens induce preventive and therapeutic anti-tumor immunity in murine p815 modelINTERNATIONAL JOURNAL OF CANCER, Issue 5 2002Yuzhang Wu Abstract The efficacy of phage display particles expressing tumor antigen P1A35-43 in inducing protective and therapeutic anti-tumor immune responses were studied. A protective immune response against a lethal progressive P815 mastocytoma tumor cell challenge was established after subcutaneous injection of phage display particles. Furthermore, the vaccine suppressed growth of pre-existing tumors. Immunization with the hybrid phage particles elicited P1A35,43 specific CTL responses and a Th1-dominated immune response with phage particle-specific secretion of IFN-, but not IL-4. Our results indicate that phage display particles might be a useful vaccine form for tumor-associated antigen epitopes in tumor immunotherapy. © 2002 Wiley-Liss, Inc. [source] A new bacteriophage, VHML, isolated from a toxin-producing strain of Vibrio harveyi in tropical AustraliaJOURNAL OF APPLIED MICROBIOLOGY, Issue 4 2000H.J. Oakey Some strains of Vibrio harveyi are known to be pathogenic for fish and many invertebrates including crustaceans. Despite their importance, their modes of virulence have yet to be fully elucidated. Here, we present a previously unreported bacteriophage extracted from a toxin-producing strain of V. harveyi isolated from moribund prawn larvae in tropical Australia. Classification into the family Myoviridae was based upon morphological characteristics (an icosahedral head, a neck/collar region and a sheathed rigid tail) and nucleic acid characteristics (double-stranded linear DNA). We have termed the bacteriophage VHML (Vibrio Harveyi Myovirus Like). VHML is a temperate bacteriophage that has a narrow host range and shows an apparent preference for V. harveyi above other vibrios (63 Vibrio isolates tested) and other genera (10 other genera were tested). The conventional methods for phage concentration and extraction of nucleic acids from phage particles were not efficient and the alternative methods that were used are discussed. [source] Regulated site-specific recombination of the she pathogenicity island of Shigella flexneriMOLECULAR MICROBIOLOGY, Issue 5 2004Harry Sakellaris Summary The she pathogenicity island (PAI) is a chromosomal, laterally acquired, integrative element of Shigella flexneri that carries genes with established or putative roles in virulence. We demonstrate that spontaneous, precise excision of the element from its integration site in the 3, terminus of the pheV tRNA gene is mediated by an integrase gene (int) and a gene designated rox (regulator of excision), both of which are carried on the she PAI. Integrase-mediated excision occurs via recombination between a 22 bp sequence at the 3, terminus of pheV and an imperfect direct repeat at the pheV -distal boundary of the PAI. Excision leads to the formation of a circular episomal form of the PAI, reminiscent of circular excision intermediates of other mobile elements that are substrates for lateral transfer processes such as conjugation, packaging into phage particles and recombinase-mediated integration into the chromosome. The circle junction consists of the pheV -proximal and pheV -distal boundaries of the PAI converging on a sequence identical to 22 bp at the 3, terminus of pheV. The isolated circle was transferred to Escherichia coli where it integrated specifically into phe tRNA genes, as it does in S. flexneri, independently of recA. We also demonstrate that Rox stimulates, but is not essential for, excision of the she PAI in an integrase-dependent manner. However, Rox does not stimulate excision by activating the transcription of the she PAI integrase gene, suggesting that it has an excisionase function similar to that of a related protein from the P4 satellite element of phage P2. [source] PY54, a linear plasmid prophage of Yersinia enterocolitica with covalently closed endsMOLECULAR MICROBIOLOGY, Issue 4 2003Stefan Hertwig Summary PY54 is a temperate phage isolated from Yersinia enterocolitica. Lysogenic Yersinia strains harbour the PY54 prophage as a plasmid (pY54). The plasmid has the same size (46 kb) as the PY54 genome isolated from phage particles. By electron microscopy, restriction analysis and DNA sequencing, it was demonstrated that the phage and the plasmid DNAs are linear, circularly permuted molecules. Unusually for phages of Gram-negative bacteria, the phage genome has 3,-protruding ends. The linear plasmid pY54 has covalently closed ends forming telomere-like hairpins. The equivalent DNA sequence of the phage genome is a 42 bp perfect palindrome. Downstream from the palindrome, an open reading frame (ORF) was identified that revealed strong DNA homology to the telN gene of Escherichia coli phage N15 encoding a protelomerase. Similar to PY54, the N15 prophage is a linear plasmid with telomeres. The N15 protelomerase has cleaving/joining activity generating the telomeres by processing a 56 bp palindrome (telomere resolution site tel RL). To study the activity of the PY54 protein, the telN -like gene was cloned and expressed in E. coli. A 77 kDa protein was obtained and partially purified. The protein was found to process recombinant plasmids containing the 42 bp palindrome. Telomere resolution of plasmids under in vivo conditions was also investigated in Yersinia infected with PY54. Processing required a plasmid containing the palindrome as well as adjacent DNA sequences from the phage including an additional inverted repeat. Regions on the phage genome important for plasmid maintenance were defined by the construction of linear and circular miniplasmid derivatives of pY54, of which the smallest miniplasmid comprises a 4.5 kb DNA fragment of the plasmid prophage. [source] Isolation of high-quality RNA from white spruce tissue using a three-stage purification method and subsequent cloning of a transcript from the PR-10 gene familyPHYTOCHEMICAL ANALYSIS, Issue 4 2003Nathalie Mattheus Abstract Isolation of PinmIII cDNA homologues from white spruce tissues required a rigorous RNA extraction protocol developed following assessment of three previously reported conifer RNA extraction protocols. Total RNA was extracted via several purification steps designed to minimize binding of phenolics to nucleic acids and was then subjected to caesium chloride ultra-centrifugation. This procedure produced consistently high-quality, intact RNA from both needles and roots with spectrophotometric ratios of approximately 2.0 for both 260/280,nm and 260/230,nm. Total RNA was obtained from the roots of cold-hardened white spruce seedlings for cDNA library construction. More than 2 million recombinant phage particles were generated from 5,µg of a poly(A)+RNA fraction, and ca. 1.3 million cDNA particles were amplified for storage. Approximately 500,000 primary recombinant clones were screened with an heterologous PinmIII cDNA sequence yielding a unique clone, picg1, that was very similar to members of the PR10 gene family. Copyright © 2003 John Wiley & Sons, Ltd. [source] Characterization of R-body genetic determinants in Caedibacter caryophilus a symbiont of Paramecium caudatum: preliminary resultsTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 2 2005MARTINA SCHRALLHAMMER Refractile inclusion bodies, called R-bodies were observed within the cells of some bacterial strains. They are protein ribbons, which are typically coiled into cylindrical structures. They are produced by members of the genus Caedibacter, gram-negative rod-shaped endosymbionts of paramecia and e.g. the free-living bacteria Hydrogenophaga taeniospiralis and Acidovorax avenae. The phylogenetic relationship even between the members of the genus Caedibacter is quite low: C. taeniospiralis belongs to the Gammaproteobacteria and is related to Francisella tularensis, C. caryophilus is affiliated with the Alphaproteobacteria and clusters with the obligate endosymbiont Holospora. In the case of C. taeniospiralis 51, the genetic determinants of R-body synthesis are located on a plasmid, whereas in other strains like 7 and 562 it looks like phage particles are involved in their production. In the present study, we investigated C. caryophilus, endosymbiont of Paramecium caudatum. Separation of C. caryophilus cells was performed by PercollÔ density gradient centrifugation. The isolated DNA was separated by pulsed-field gel electrophoresis and it was possible to visualize several bands referring to one or more extrachromosomal elements. A small gene library of these extrachromosomal elements was constructed and we already identified transposition-related genes; interestingly, similar genes were reported also on the plasmid of C. taeniospiralis 51. [source] Cover Picture: ChemBioChem 1/2003CHEMBIOCHEM, Issue 1 2003Sachdev S. Sidhu Dr. The cover picture shows a highly diverse peptide library displayed on cylindrical bacteriophage particles that also contain the cognate DNA. Libraries such as this can be used to identify specific ligands for essentially any protein of interest. Here, phage particles displaying peptides that bind vascular endothelial growth factor are selected from amongst a sea of billions of random sequences. Further information can be found in the Review by Sidhu and co-workers on p. 14 ff. (We thank David Wood and Christian Wiesmann for help in the preparation of the cover picture.) [source] | |||||||||||||