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Detection of Clostridium botulinum by Multiplex PCR in Foods and Feces
Time of Update: 2021-02-16
Clostridium botulinum is a diverse group of anaerobic spore-forming organisms that produce lethal botulinum neurotoxins (BoNT) during their growth.
The most common forms of human botulism include the classical foodborne botulism due to ingestion of BoNT preformed in food, and infant botulism due to spore germination, growth, and toxin production in the infant’s intestine.
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Identification of Protein Complexes from Filamentous Fungi with Tandem Affinity Purification
Time of Update: 2021-02-16
Tandem affinity purification (TAP) allows the enrichment of native protein complexes from cell extracts under mild conditions.
We codon-optimized tags and established TAP, previously not applicable to filamentous fungi, for the model organismAspergillus nidulans.
nidulans which can also be adapted to other filamentous fungi.
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PCR-Derived Methods for the Identification of Trichinella Parasites from Animal and Human Samples
Time of Update: 2021-02-16
Trichinella worms (family: Trichinellidae; phylum: Nematoda) are parasites that mainly infect mammals, including humans, although they have been found in birds and, recently, in African crocodiles ( 1 , 2 ).
The main reservoir is represented by carnivores with cannibalistic and scavenger behavior.
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Flow Cytometry in Environmental Microbiology: A Rapid Approach for the Isolation of Single Cells for Advanced Molecular Biology Analysis
Time of Update: 2021-02-16
The isolation and subsequent characterization of microbial cells from within environmental samples is a difficult process.
We describe here the combination of fluorescent in situ hybridization and cell sorting for the detection and isolation of Candidate Division TM7 bacteria from an enriched soil sample.
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Selectable Markers for Eukaryotic Cells
Time of Update: 2021-02-16
The transfer of DNA sequences into a population of cells can rarely, if ever, be achieved with 100% efficiency.
If the transferred sequences do not confer a selective growth advantage, it is essential to use selection for transduced cells.
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Using an Advanced Microfermentor System for Strain Screening and Fermentation Optimization
Time of Update: 2021-02-16
To maximize the bioproduction of a selected strain, bioreaction (fermentation) conditions need to be optimized.
Bench-scale fermentor experiments (>0.5 L) are well controlled under designed conditions and provide high-quality data, but they are also very time- and cost-consuming.
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Electronic Access Mycobacterium tuberculosis Sequence Data
Time of Update: 2021-02-16
This information is obviously of great benefit to researchers in the field, but the sheer volume of data often makes it inaccessible.
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Metabolomic Investigation of Methicillin-Resistant Staphylococcus aureus
Time of Update: 2021-02-16
An efficient and accurate method to measure the intracellular metabolites of a specific microbial species is a key prerequisite for metabolome analysis.
This protocol is demonstrated to be an efficient method for analyzing the intracellular metabolites ofS.
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Mesophilic Aerobic Microorganisms
Time of Update: 2021-02-16
The mesophilic microorganism is ones of the more general and extensively microbiological indicators of food quality, ( 1 ) indicating the adequacy of temperature and sanitation control during process
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Virus Genome Detection by the Polymerase Chain Reaction (PCR)
Time of Update: 2021-02-16
Since the polymerase chain reaction ( PCR ) technique was developed in 1986 ( 1 ), it has found wide application in molecular biology and is now regarded as an irreplaceable tool in many laboratories
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Fungal Genomic DNA Extraction
Time of Update: 2021-02-16
Overview High throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes, half full with liquid media (500uL), with a hole punched through the top for aeration; or if greater volumes are required, (b) peteri plates containing 10mL liquid media.
Transfer top phase to fresh 1.5mL microfuge tube, add 1/2 vol.
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Genetic Modification in Bacillus subtilis for Production of C30 Carotenoids
Time of Update: 2021-02-16
This chapter describes a method to synthesize C30 carotenoids inBacillus subtilis , which is generally recognized as a safe (GRAS) organism.
aureus crtM andcrtN genes intoB.
Thus, carotenoid biosynthesis genes ofS.
subtilis in order to construct a safe organism producing C30 carotenoids.
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Construction of Phage Antibody Repertoires from the Blood of West Nile Virus-Infected Donors
Time of Update: 2021-02-16
A method for the construction of West Nile virus immune donor antibody repertoires is described.
The PCR fragments are cloned in a phage display vector to construct a repertoire that can be used in panning procedures to identify many unique monoclonal antibodies.
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High-Level Periplasmic Expression and Purificationof scFvs
Time of Update: 2021-02-16
The isolation of recombinant antibodies by phage display naturally leads to experiments to evaluate their biological and immunological properties. Although crude preparations may have their value in
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Directed Module Detection in a Large-Scale Expression Compendium
Time of Update: 2021-02-16
In this chapter, we illustrate how the web services COLOMBOS and DISTILLER can be used to identify condition-dependent coexpression modules by exploring compendia of public expression data.
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Purification and Characterization of MHC Class II Containing Organelles in Mouse Bone-Marrow-Derived Dendritic Cells
Time of Update: 2021-02-16
However, data from cell-fractionation studies, predominantly those using electrophoresis techniques, show that in professional APC, a class of class II-containing endosomes that is specialized for antigen processing may exist.
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Detection of Human Caliciviruses and Astroviruses in Stools by RT-PCR
Time of Update: 2021-02-16
Human caliciviruses (HuCVs) and astroviruses are single-stranded RNA viruses that cause acute gastroenteritis in humans.
Astroviruses include at least seven antigenic types ( 4 ).
In both families, differences in antigenic types appear to correspond to differences in genetic groups.
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Isolation of Mouse Thymic Dendritic Cells
Time of Update: 2021-02-16
The method described in this chapter for the isolation of mouse thymic dendritic cells (DC) is an optimization of our previously published methods (1 ,2 ) and involves the following major steps: 1.
4. Positive selection of DCs by magnetic cell sorting (MACS).
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Electroporation of Francisella tularensis
Time of Update: 2021-02-16
In North America, tularemia presents as an acute febrile lymphadenitis that can progress into a life-threatening pneumonic illness ( 1 ).
In Europe and Asia, tularemia is often acquired from consumption of contaminated water or inhalation of contaminated dust.
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In Vitro and In Vivo Interactions of Nonpathogenic Bacteria With Immunocompetent Cells
Time of Update: 2021-02-16
Specific components of the intestinal microflora, includingLactobacilli andBifidobacteria , have been associated with beneficial effects on the host, such as promotion of gut maturation and integrity, antagonisms against pathogens, and immune modulation.
To date, the contribution of the intestinal epithelial cell in the first-line defense against pathogenic bacteria and microbial antigens has been recognized.
