Exopolysaccharides (EXPs) of Staphylococcus aureus are associated with virulence in animal models. An EXP from the S. aureus strain Smith diffuse was previously detected in 64.3% of S. aureus clinical isolates. EXP was isolated from culture supernatants of this strain after DNAase, RNAase, phosphodiesterase I and lysostaphin treatment, and was further purified by cation-exchange and molecular-sieve chromatography. Isoelectric focusing revealed a pI of 3.6 for the EXP while the pI of teichoic acid was less than 2.7. Crossed immunoelectrophoresis with homologous Smith diffuse antisera indicated that the EXP contained two immunological components. A major precipitin line persisted after the antisera had been absorbed with the non-EXP-producing variant strain, Smith compact, while the second component was removed. Tandem immunoelectrophoresis also demonstrated that the EXP was distinct from teichoic acid. The EXP contained 2-amino-2-deoxyglucuronic acid, glucose, mannose and galactose. No fatty acids or nucleic acids were present and total protein content was less than 2%. Teichoic acid could not be demonstrated in the EXP, thus further substantiating the immunological studies. S. aureus EXP isolated by the present method can be used for further serological and virulence studies.
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http://dx.doi.org/10.1099/00221287-133-2-431 | DOI Listing |
J Gen Virol
January 2025
Biochemistry Program, The University of the South, Sewanee, TN, USA.
The murine hepatitis virus (MHV) is an important model system for studying coronavirus (CoV) molecular and cell biology. Despite this, few reagents for MHV are available through repositories such as ATCC or Addgene, potentially limiting the widespread adoption of MHV as a tractable model system. To overcome some challenges inherent in the existing MHV reverse genetics systems, we developed a plasmid-launched transformation-associated recombination (TAR) cloning-based system to assemble the MHV (strain A59; MHV-A59) genome.
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Department of Orthopaedic Surgery, University of Utah, Salt Lake City, Utah, USA.
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Nat Cell Biol
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Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO, USA.
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Department of Molecular Biology, School of Biological Sciences, University of California at San Diego, 9500 Gilman Dr, La Jolla, CA 92093-0116, USA.
Insertions of the transposable element IS5 into its target sites in response to stressful environmental conditions, DNA structures, and DNA-binding proteins are well studied, but how the genomic contexts near IS5's native loci impact its transpositions is largely unknown. Here, by examining the roles of all 11 copies of IS5 within the genome of strain BW25113 in transposition, we reveal that the most significant copy of IS5 is one nested within and oriented in the same direction as the gene, while two other copies of IS5 harboring point mutations are hardly transposed. Transposition activity is heavily reliant on the upstream promoter that drives IS5 transposase gene , with more transpositions resulting from greater promoter activity.
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University Coimbra, Centre for Mechanical Engineering, Materials and Processes (CEMMPRE), Department of Mechanical Engineering, 3030-788 Coimbra, Portugal.
The stop-hole technique is a well-known strategy to extend the fatigue life of cracked components. The ability to estimate fatigue life after the hole is important for safety reasons. The objective here is to develop strategies for the accurate prediction of initiation and propagation life ahead of the stop-hole.
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