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Immunological Methods for the Detection of Human Cytomegalovirus
Time of Update: 2021-02-22
In addition, HCMV has been implicated as a co-etiological agent in cervical cancer ( 2 ) and has been found associated with a wide range of other tumors ( 1 ).
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Transformation of Candida maltosa by Electroporation
Time of Update: 2021-02-22
Candida species are diploid or aneuploid in their genetic constitution and have proven to be difficult organisms to study in terms of their genetics and molecular biology.
Moreover, electroporation permits the introduction of DNA into organisms that are refractory to other transformation techniques ( 2 ).
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Reproducible Quantitative PCR of Mitochondrial and Nuclear DNA Copy Number Using the LightCycler
Time of Update: 2021-02-22
Recent developments inPCR fluorimetry have allowed for quick quantification of target molecules. Before the invention of fluorimetric quantitative PCR, researchers who wanted to quantify the amount
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Monitoring Microbial Activities Using Membrane Inlet Mass Spectrometry
Time of Update: 2021-02-22
Currently used methods include manometric techniques, microsensors, chemical assays, gas chromatography and high-performance liquid chromatography, but all have their limitations and usually require substantial disruption to the environment being studied.
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Biosynthesis, Extraction, Purification, and Analysis of Trisporoid Sexual Communication Compounds from Mated Cultures of Blakeslea trispora
Time of Update: 2021-02-22
The zygomyceteBlakeslea trispora produces high amounts of the general zygomycete β-carotene-derived sexual signal compounds, the trisporoids.
The effect of the extraction and activity of the isolated compounds is best tested physiologically, exploiting the ability of trisporoids to induce the formation of sexually committed hyphae, the zygophores, in other zygomycete species.
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Neurosporaxanthin Production by Neurospora and Fusarium
Time of Update: 2021-02-21
The orange pigmentation of the ascomycete fungiNeurospora andFusarium is mainly due to the accumulation of neurosporaxanthin, a carboxylic apocarotenoid whose possible biotechnological applications have not been investigated.
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Inducible Expression Systems for Mycobacteria
Time of Update: 2021-02-21
A wide variety of inducible expression systems have been designed for Gram-negative bacteria, but adapting these systems to phylogenetically distinct species, such as mycobacteria, has proved notoriously difficult.
Although comparatively few genetic tools are available for these organisms, those that do exist are more likely to be adaptable for use in mycobacteria.
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In Vivo Assays of Langerhans Cell Migration
Time of Update: 2021-02-21
The skin is an immunologically active tissue; the integrated skin immune system comprises epidermal Langerhans cells (LC), dermal dendritic cells (DC), tissue macrophages, mast cells, and T lymphocyt
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Generation of Astaxanthin Mutants in Xanthophyllomyces dendrorhous Using a Double Recombination Method Based on Hygromycin Resistance
Time of Update: 2021-02-21
In this chapter is described a method that allows the double gene deletions of the two copies of a gene from a diploid organism, a wild-type strain of theXanthophyllomyces dendrorhous yeast, using hygromycin B resistance as the only selection marker.
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Application of Microwave-Assisted Extraction for the Analysis of Dithiocarbamates in Food Matrices
Time of Update: 2021-02-21
Quantification is based on external standard calibration curves made with either standard solutions of individual DMDTCs (thiram, ziram) or EBDTCs (maneb, zineb, mancozeb), which are processed as the field samples, or using standard solutions of CS 2 made in isooctane.
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Food-Grade Corynebacteria for Enzyme Production
Time of Update: 2021-02-21
Using genetic manipulation techniques, several corynebacteria strains expressing genes coding for hydrolytic enzymes or proteins have been constructed in different laboratories.
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Comparative Genome Analysis of Clostridium difficile Using DNA Microarrays
Time of Update: 2021-02-21
However, to fully exploit this opportunity, robust phylogenetic methods on a diverse set of characterised strains are required to provide a reference evolutionary framework to studyC.
Traditional phylogenetic classification of bacteria to study evolutionary relatedness is based on the characterisation of a limited number of genes, rRNA or signature sequences.
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Animal Models in Drug Development for MRSA
Time of Update: 2021-02-21
In this way animal models of infection can be a highly valuable and predictive bridge between in vitro drug discovery and early clinical evaluation.
This chapter will describe the advantages, methods, and outcome measurements of the rodent models most used in drug discovery forS.
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The Use of Monoclonal Antibodies and Lectins to Identify Changes in Viral Glycoproteins That are Influenced by Glycosylation: The Case of Human Respiratory Syncytial Virus Attachment (G) Glycoprotein
Time of Update: 2021-02-21
Given the role of the G glycoprotein in RSV antigenicity, we also describe procedures based on Western blotting to determine the effect of G protein glycosylation changes on reactivity with human sera.
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Bacteriocin Production Process by a Mixed Culture System
Time of Update: 2021-02-21
Antimicrobial proteins and peptides produced by bacteria, termed bacteriocin, are widely acknowledged to be important contributors to those organisms that survive dominate or die in microbial ecosystem such as our food supply or digestive tract.
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Measurement of Tumor Necrosis Factor (TNF) Lysis of Adenovirus-Infected Cells
Time of Update: 2021-02-21
To minimize or prevent the spread of an acute virus infection, the antiviral immune response must detect and lyse virus-infected cells before virus replicates or is released from the host cell.
The immune response has developed both innate and specific responses to meet this objective.
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Quantification of Host and Phage mRNA Expression During Infection Using Real-Time PCR
Time of Update: 2021-02-21
Real-time, or quantitativePCR , is a valuable technique useful in bacteriophage research to quantify the abundance of phage or host gene transcripts.
This chapter should also be useful to those wishing to amplify genes from other types of templates such as metagenomic DNA or RNA extracted either from filtered samples or from agarose gels.
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Escherichia coli Electrotransformation
Time of Update: 2021-02-21
With electrotransformation, as many as 80% of the cells receive exogenous DNA ( 1 ).
Chemical and natural methods of DNA transfer into bacteria are also effective for transforming bacteria, but they are more cumbersome and time-consuming, and generally less efficient than electrotransformation.
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Fit-for-Purpose Quenching and Extraction Protocols for Metabolic Profiling of Yeast Using Chromatography-Mass Spectrometry Platforms
Time of Update: 2021-02-21
Methods to sample the exometabolome and to quench metabolism and extract intracellular metabolites for the model eukaryoteSaccharomyces cerevisiae are presented here.
For endometabolome studies, fast quenching of intracellular metabolism is performed using a 60:40 (v/v) methanol:aqueous ammonium hydrogen carbonate solution at –48�C.
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Measurement of Cytokines in Synovial Fluid
Time of Update: 2021-02-21
Over the last 20 years, advances in the knowledge of cytokine biology have brought a new dimension to the understanding of pathogenic events in human chronic arthritides, showing promise for future therapeutic approaches.
It is beyond the scope of this chapter to describe in detail the available evidence concerning the role of cytokines in these diseases.
