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Measurement of Macrophage-Mediated Cytolysis of Adenovirus-Infected Cells
Time of Update: 2021-02-01
Macrophages are well-known for their ability to serve as phagocytes, ingesting and destroying microorganisms such as bacteria, and for their function in antigen presentation. Less appreciated, perhap
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Functional Gene Arrays for Analysis of Microbial Communities on Ocean Platform
Time of Update: 2021-02-01
The Environmental Sample Processor (ESP) minimizes this time difference by housing a robotic system placed in the ocean for a long period of time that can collect a small sample, concentrate theDNA , run a microarray, and take a picture of the array before sending the data ashore to be evaluated by a researcher.
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Analytical Ultracentrifugation of Chromatin
Time of Update: 2021-02-01
Consequently, to biochemically characterize chromatin structure/function relationships in vitro, one must be able to analyze both the intramolecular conformational dynamics and intermolecular interactions of an exceedingly complex macromolecular assembly (e.g., a 12-mer nucleosomal array containing one H1 molecule per nucleosome consists of >100 proteins and 2400 bp of DNA, has a molecular mass of approx 3.5�106 D, yet represents only roughly one millionth of an intact eukaryotic chromosome.) Fig. 1.
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Detection of Enteroviruses
Time of Update: 2021-02-01
Nowadays, molecular biology techniques allow the use of very sensitive and specific reverse-transcription polymerase chain reaction (RT- PCR ) procedures to detect enteroviruses.
Finally, protocols for molecular detection of enterovirus are described, including procedures for conventional, nested, and real-timeRT-PCR .
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Sequence Analysis of the Variable Number Tandem Repeat in Staphylococcus aureus Protein A Gene: spa Typing
Time of Update: 2021-02-01
VNTR analysis is currently widely used to sub-speciate many bacterial, fungal, and viral pathogens and has facilitated a number of molecular epidemiology studies.
In this chapter, we focus onspa typing which is based on sequence analysis of VNTRs in the polymorphic X region of theStaphylococcus aureus protein A geneStaphylococcus aureus .
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Large-Scale Purification and Crystallization of Adenovirus Hexon
Time of Update: 2021-02-01
This chapter provides a protocol for the large-scale purification of adenovirus type 2 and 5 virions and the soluble major coat protein hexon.
The infected cells are lysed, virions and hexon are separated by centrifugation, and the protein is then further purified by anion exchange chromatography.
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Chromatin Immunoprecipitation to Study the Binding of Proteins to the Adenovirus Genome In Vivo
Time of Update: 2021-02-01
Repeated DNA sequences in the packaging domain contribute to viral DNA encapsidation, and several viral proteins bind to these repeats when analyzed using in vitro DNAprotein-binding assays.
The technique is easily adaptable to study the interaction of any viral or cellular protein to Ad DNA or to cellular genomic DNA sequences.
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Protein Disulfide Bond Formation in the Periplasm: Determination of the In Vivo Redox State of Cysteine Residues
Time of Update: 2021-02-01
Many proteins secreted to the bacterial cell envelope contain cysteine residues that are involved in disulfide bonds.
Monitoring the in vivo redox state of cysteine residues, i.e., determining whether those cysteines are oxidized to a disulfide bond or not, is therefore required to fully characterize the function and structural properties of numerous periplasmic proteins.
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Identification of Mycoplasmas by Immunofluorescence
Time of Update: 2021-02-01
Another diagnostic application of immunofluorescence is the detection of mycoplasma antibodies ( 2 ), but the topic falls outside the scope of this chapter.
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The syringe
Time of Update: 2021-02-01
Every year from mid-March to early April, pro-fish swims to the coast to lay eggs.
The amount of eggs is 1 to 20,000.
The herring herd of bait is also not far from the coast.
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Bacterial Molecular Networks: Bridging the Gap Between Functional Genomics and Dynamical Modelling
Time of Update: 2021-02-01
This introductory review synthesizes the contents of the volumeBacterial Molecular Networks of the seriesMethods in Molecular Biology .
This volume gathers 9 reviews and 16 method chapters describing computational protocols for the analysis of metabolic pathways, protein interaction networks, and regulatory networks.
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Adhesion Ability of Lactobacillus to Vaginal Epithelial Cells: Study by Microbiological Methods
Time of Update: 2021-02-01
Adhesion of lactobacilli to the epithelium has been described as the first step in the formation of a barrier to prevent undesirable microbial colonization ( 1 ); consequently, it has been defined as
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Genomic Footprinting Using Nucleases
Time of Update: 2021-02-01
Investigations of protein/ DNA interactions in vitro may not have relevance to a living cell system.
To analyze events occurring at the DNA level in a living cell, Church et al.
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Expression of Single-Chain Fv Fragments in E. coli Cytoplasm
Time of Update: 2021-02-01
The most frequently used approach to produce single-chain Fv fragments (scFv) and Fab inEscherichia coli is to express them in the periplasm of the bacteria.
coli strains or hyperstable scFvs.
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Refinement of the Hamster Model of Clostridium difficile Disease
Time of Update: 2021-02-01
The Golden Syrian hamster is widely regarded as the most relevant small animal model ofClostridium difficile disease as oral infection of animals pre-treated with antibiotics reproduces many of the symptoms observed in man.
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Controlled Expression of Recombinant Genes and Preparation of Cell-Free Extracts in Yeast
Time of Update: 2021-02-01
Additionally, complex biochemical processes such as transcription andDNA repair can be studied in yeast cell-free extracts in vitro, which benefit greatly from a large collection of well-defined mutant strains.
Controlled gene expression and preparation of cell-free extracts are important techniques in the yeast system.
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The Detection of Enteroviruses in Water and Associated Materials Using the Polymerase Chain Reaction
Time of Update: 2021-02-01
Although not a cause of gastroenteritis, enteroviruses may cause disease including paralysis, meningitis, myocarditis, and cardiomyopathy, and less severe infections, such as, colds and fever, mainly in young children, They are therefore a public health concern, and in order to make proper evaluation of their significance, in water techniques are required that are rapid and reliable and can accommodate large numbers of samples in one test batch.
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Molecular Methods for Detecting Ulcerogenic Strains of H. pylori
Time of Update: 2021-02-01
These characteristics can be divided into two groups: first, those relating to vacuolating cytotoxin activity ( 1 , 2 ), and differences in the gene encoding the cytotoxin,vacA ( 3 ); second, those relating to the cytotoxin-associated gene,cagA ( 4 , 5 ).
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PCR-Based Detection of Coxiella burnetii from Clinical Samples
Time of Update: 2021-02-01
burnetii is performed in cell culture, animals or embryonated chicken eggs, however, the procedure is time-consuming and hazardous and therefore restricted to specialized laboratories.
A PCR assay designated Trans-PCR, which targets a repetitive, transposon-like element ( 5 – 8 ) proved to be specific and sensitive.
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Overcoming Inhibition in Real-Time Diagnostic PCR
Time of Update: 2021-02-01
PCR is an important and powerful tool in several fields, including clinical diagnostics, food analysis, and forensic analysis.
In order to overcome the problems caused by PCR inhibitors, all the steps leading up to DNA amplification must be optimized for the sample type in question.
