Inclusion Bodies of Recombinant OmpF Porin from Yersinia pseudotuberculosis : Properties and Structural Characterization

AbstractMature pore-forming OmpF protein from the outer membrane ofYersinia pseudotuberculosis was expressed inEscherichia coli in the form of inclusion bodies (IBs) under different cultivation conditions. The properties and structural organization of the IBs as well as the structure of the recombinant porin (rOmpF) solubilized from the IBs were investigated using electron microscopy, dynamic light scattering, optical spectroscopy, and specific hydrophobic dyes. The size, shape, and stability of the IBs under denaturing solutions were determined. It was found that the IBs were readily soluble in SDS and more resistant to urea. Dissolution of the IBs in both denaturing agents led to formation of a heterogeneous in size population of oligomeric particles. The IBs contained an intermediate form of the rOmpF with native-like secondary structure and elements of tertiary structure, which was able to penetrate a lipid bilayer and adopt a functionally active conformation. There were no significant differences in the properties and structure between the examined IBs formed at different concentrations of the inducer (IPTG). However, the content of amyloids in the IBs increased with increasing concentration of the inducer. These results contribute to the development of new approaches for the production of active proteins from IBs, as well as biologically and functionally active IBs.
Source: Biochemistry (Moscow) - Category: Biochemistry Source Type: research

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AbstractIn acute gastroenteritis (AGE), identification of the infectious agent is important for patient management. Since symptoms do not reliably identify the agent, microbiological diagnostics are important. Conventional methods lack sensitivity and often take days. Multiplex PCR panels offer fast and sensitive alternatives. Our aim was to assess the performance of the new QIAstat Gastrointestinal Panel (GIP) detecting 24 different gastroenteric pathogens from stool in Cary-Blair transport medium (Adenovirus F 40/41, Astrovirus, Norovirus GI/GII, Rotavirus A, Sapovirus,Campylobacter spp.,Clostridium difficile,Plesiomonas...
Source: European Journal of Clinical Microbiology and Infectious Diseases - Category: Microbiology Source Type: research
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Source: Transfusion - Category: Hematology Authors: Tags: Transfusion Source Type: research
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Source: Journal of Organometallic Chemistry - Category: Chemistry Source Type: research
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Source: Environmental Pollution - Category: Environmental Health Authors: Tags: Environ Pollut Source Type: research
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Source: GIDEON blog - Category: Databases & Libraries Authors: Tags: Ebooks Epidemiology ProMED Source Type: blogs
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Source: Pediatric Emergency Care - Category: Emergency Medicine Tags: Illustrative Cases Source Type: research
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Source: Current Topics in Microbiology and Immunology - Category: Microbiology Tags: Curr Top Microbiol Immunol Source Type: research
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Source: Am J Trop Med Hyg - Category: Infectious Diseases Authors: Tags: Am J Trop Med Hyg Source Type: research
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Source: Journal of Microbiology, Immunology and Infection - Category: Microbiology Source Type: research
CONCLUSION: Altogether, our data suggested that loxapine might suppress intracellular bacteria through inhibiting of bacterial efflux pumps. In light of its unique activity, loxapine represents a promising lead compound with translational potential for the development of a new antibacterial agent against intracellular bacteria. PMID: 31239204 [PubMed - as supplied by publisher]
Source: Journal of Microbiology, Immunology, and Infection - Category: Microbiology Authors: Tags: J Microbiol Immunol Infect Source Type: research
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