-
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.
-
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.
-
Assay of Antibacterial Activities of the Bactericidal/Permeability-Increasing Protein in Natural Biological Fluids
Time of Update: 2021-02-01
This process is driven by microbe-specific signals, including highly conserved and chemically unique microbial (envelope) components, such as Gram-negative bacterial lipopolysaccharides (LPS) ( 1 , 2 ).
-
Metabolic Pathway Determination and Flux Analysis in Nonmodel Microorganisms Through 13C-Isotope Labeling
Time of Update: 2021-02-01
The five key procedures are: (1) growing cells using labeled substrates, (2) measuring extracellular metabolite and biomass component, (3) analyzing isotopic labeling patterns in amino acids and central metabolites using gas chromatography-mass spectrometry, (4) tracing13 C carbon transitions in metabolites and discovering new pathways, and (5) estimating flux distributions based on isotopomer constraints.
-
Molecular Imprinted Solid-Phase Extraction for Cleanup of Chlorinated Phenoxyacids From Aqueous Samples
Time of Update: 2021-02-01
SPE procedures involving molecular imprinted polymers (MISPE, molecular imprinted solid-phase extraction) have been proposed as a valid substitute for immunoaffinity procedures because of their simplicity of preparation and high stability to extreme chemical conditions.
-
Cell-Free Transcription of SV40
Time of Update: 2021-02-01
The development of soluble cell-free systems that accurately transcribe genes has provided a means to investigate the mechanisms of eukaryotic transcription.
Our laboratory has been investigating the molecular mechanisms by which the genes of SV40 are transcriptionally regulated.
-
Use of Commercially Available ELISA Kits for Detection of Foodborne Pathogens
Time of Update: 2021-02-01
The IgG molecule is by far the most common, and its structure is shown in Fig. 1 .
This unique recognition of an antigen by an antibody is the principle that makes the immunoassay such a powerful diagnostic tool.
-
Display and Selection of scFv Antibodies on HEK-293T Cells
Time of Update: 2021-02-01
Flow cytometry enhances the screen’s sensitivity thereby allowing us to isolate high-affinity scFvs. Using human cell display, one could isolate and engineer scFvs, single domains, Fabs, or whole IgGs for increased affinity and other biological functions.
-
Assembly of Bacterial Outer Membrane Proteins
Time of Update: 2021-02-01
Various methods that are routinely used to study the subcellular localization of membrane proteins in wild-type Gram-negative bacteria fall short in genetic studies addressing the biogenesis of outer membrane proteins (OMPs).
-
The Use of Temperature-Sensitive Plasmids in Mycobacteria
Time of Update: 2021-02-01
The construction of allelic exchange mutants is one of the major strategies to decipher the function of a defined gene.
smegmatis mutants and to test whether or not a gene is essential.
-
Use of Pichia pastoris for Production of Recombinant Cytokines
Time of Update: 2021-02-01
A variety of expression systems have been used to produce recombinant proteins, includingEscherichia coli ,Pichia pastoris , baculovirus, and poxvirus systems.
pastoris offers the potential for high yields of biologically active recombinant cytokines at relatively low production costs.
-
Analysis of Yeast Lipids
Time of Update: 2021-02-01
The precise quantitative determination of the different lipid classes in mutant cells is key to understand the possible role of the respective gene product in lipid homeostasis. In this chapter, we d
-
Conjugation of Haptens
Time of Update: 2021-02-01
In the design of hapten conjugates, consideration must be given to the hapten, the carrier, the coupling strategy, and the hapten density because the amount of hapten attached to the carrier influences the strength of the immune response directed toward the newly created antigenic determinant.
-
Analysis of Endocrine Disruptor Pesticides in Adipose Tissue Using Gas Chromatography-Tandem Mass Spectrometry: Assessment of the Uncertainty of the Method
Time of Update: 2021-02-01
The results show that the sources of largest uncertainty are those relative to the balance calibration, from the gravimetric step, and both the relative uncertainty associated with the recovery and the intermediate precision of the method.
-
Immunologic Methods and Correlates of Protection
Time of Update: 2021-02-01
Serum neutralizing antibodies (nAbs) occur after natural RV infection in children, and are serotype-specific ( 4 , 6 ).
-
Dissecting the Cell Surface
Time of Update: 2021-02-01
The bacterial cell surface is an important structure as it mediates interactions with the external environment.
In the case of pathogens likeClostridium difficile , the cell wall and its components also have to mediate interactions with the host cells and their products.
-
Measuring In Vivo Signaling Kinetics in a Mitogen-Activated Kinase Pathway Using Dynamic Input Stimulation
Time of Update: 2021-02-01
We can measure a property we termed pathway bandwidth to putin vivo bounds on the kinetics of the mitogen-activated protein kinase (MAPk) signaling cascade inSaccharomyces cerevisiae that responds to hyperosmotic stress [the High Osmolarity Glycerol (HOG) pathway].
-
Carotenogenic Microorganisms: A Product-Based Biochemical Characterization
Time of Update: 2021-02-01
At least four particular carotenoids (Fig. 1 ) are fully exploited for commercial applications because their production was consolidated through chemical synthesis: β-carotene (C 40 double cyclic ends), canthaxanthin (diketo-β-carotene), astaxanthin (dihydroxy-diketo-β-carotene), and apocarotenoic acid as its ethyl ester (C 32 ; single cyclic end).
-
The Application of Bacterial Luciferase as a Reporter Gene in Mycobacteria
Time of Update: 2021-02-01
Bioluminescence is the product of two distinct enzymes, firefly luciferase and bacterial luciferase.
The reaction can be written as follows: FMNH 2 + RCHO + O 2 → FMN + H 2 O + RCOOH + Light
-
Herpesviruses: A Brief Overview
Time of Update: 2021-02-01
Each virus is closely associated with its main host species.
Interestingly, many herpesviruses seem to be entirely avirulent within their original hosts.
However, many herpesviruses are associated with various degrees of disease in their original host.
