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Mouse Models for Studying Orthopoxvirus Respiratory Infections
Time of Update: 2021-02-14
The popular small-animal orthopoxvirus models employ the inbred mouse as the host, the respiratory tract as the site of virus inoculation, and orthopoxviruses—vaccinia, cowpox, and ectromelia viruses—as surrogates for variola virus.
This chapter describes an ectromelia virus respiratory infection model in the mouse.
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Flow Cytometric Detection of Adenoviruses and Intracellular Adenovirus Proteins
Time of Update: 2021-02-14
Previous bioassays used to quantitate adenoviruses (such as the plaque assay or fluorescent focus assay) are time-consuming and subjective in their interpretation.
Here we describe a flow cytometric method that eliminates these disadvantages and provides a quantitative and reliable method of focus-forming unit (FFU) assay.
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Gene Replacement and Transposon Delivery Using the Negative Selection Marker sacB
Time of Update: 2021-02-14
Gene replacement and transposon mutagenesis are two complementary tools that have been widely used to perform genetic studies in various living organisms. In mycobacteria, and especially in theMycoba
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Use of a Yeast-Based Membrane Protein Expression Technology to Overexpress Drug Resistance Efflux Pumps
Time of Update: 2021-02-14
We have developed a heterologous membrane protein expression system that can be used to study the structure and function of these proteins in the non-pathogenic, genetically stable, and versatile eukaryotic model organism,Saccharomyces cerevisiae .
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Methods for the Detection and Concentration of Bacteriocins Produced by Lactic Acid Bacteria
Time of Update: 2021-02-14
Lactic acid bacteria (LAB), used for centuries by man to preserve food, produce a wide variety of antagonistic compounds, including lactic acid, hydrogen peroxide, and bacteriocins.
Bacteriocins are antimicrobial peptides that are bactericidal toward bacteria taxonomically close to the producer ( 1 ).
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Quantitative Analysis of Periodontal Pathogens by ELISA and Real-Time Polymerase Chain Reaction
Time of Update: 2021-02-14
The development of analytical methods enabling the accurate identification and enumeration of bacterial species colonizing the oral cavity has led to the identification of a small number of bacterial pathogens that are major factors in the etiology of periodontal disease.
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Intracellular Expression of Recombinant Antibody Fluorescent Protein Fusions for Localization of Target Antigens in Schizosaccharomyces pombe
Time of Update: 2021-02-14
We show here that recombinant antibody fluorescent protein fusions can be used to monitor the localization of intracellular antigens in fixed or living cells.
The antibody binds to its target antigen without inhibiting protein function, allowing visualization of its intracellular location in fixed or living cells.
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Cloning, Expression, and Functional Analysis of Patient-Derived Hepatitis C Virus Glycoproteins
Time of Update: 2021-02-14
Furthermore, we describe a method to generate retroviral nucleocapsid pseudotyped with HCV E1E2 of diverse genotypes (HCVpp) whereby infectivity of the retroviral particle is conferred by HCV glycoproteins.
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The Chemical Identification and Analysis of Aspergillus nidulans Secondary Metabolites
Time of Update: 2021-02-14
The recent genomic sequencing of severalAspergillus species has revealed that many secondary metabolite gene clusters are apparently silent under standard laboratory conditions.
A straightforward, reliable method to purify and characterize new metabolites therefore should be useful.
Finally, theNMR characterization of previously unknownA.
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Recombinant Microorganisms for the Biosynthesis of Glycosylated Antitumor Compounds
Time of Update: 2021-02-14
The glycosy-lation pattern of these complex metabolites can be altered by chemical synthesis or synthetic modification of intermediates usually produced via fermentation and by the use of genetically engineered recombinant microorganisms and biosynthetic genes for in vivo experiments through combinatorial biosynthesis.
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Immunofluorescence Microscopy for the Detection of Surface Antigens in Methicillin-Resistant Staphylococcus aureus (MRSA)
Time of Update: 2021-02-14
Immunofluorescence microscopy is a widely used laboratory method which allows detection and visualization of specific antigens.
Here, we apply the method to follow antigen expression on the surface of MRSA cells over time in in vivo infection models.
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Chromatin Immunoprecipitation Analysis in Filamentous Fungi
Time of Update: 2021-02-14
Chromatin immunoprecipitation (ChIP) is used to map the interaction between proteins andDNA at a specific genomic locus in the living cell.
After reversal of crosslinks and DNA purification locus-specific quantitativePCR is used to determine the amount of DNA that was associated with the target at a given time point and experimental condition.
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A Review of Surveillance Networks of Food-Borne Diseases
Time of Update: 2021-02-14
To monitor the impact of these food-borne pathogens on human health, systems giving further information are required; a food-borne disease surveillance program is an essential part of a food safety program.
This chapter is a short overview of various surveillance networks specializing in food-borne diseases.
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Identification of Diagnostic Proteins in Mycobacterium avium subspecies paratuberculosis by a Whole Genome Analysis Approach
Time of Update: 2021-02-14
paratuberculosis now complete, we have taken a different strategy to identify novel proteins that are present uniquely inM.
paratuberculosis with other sequenced mycobacterial genomes, we identified a collection of more than 90 genes that are present uniquely inM.
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Large DNA Transformation in Plants
Time of Update: 2021-02-14
Large DNA (>100 kb) transformation methods have been reported in tomato usingAgrobacterium -mediated transformation ( 1 ) and tobacco using biolistic bombardment ( 2 ).
BIBAC vector (used in tomato) is capable of transferring large DNA fragments fromAgrobacterium into plants.
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In Vitro Transcription: Probing the Molecular Functions of Adenovirus Regulatory Proteins
Time of Update: 2021-02-14
These E1A 243 R functions are encoded within multiple protein domains that can transcriptionally activate or repress cellular genes involved in the regulation of cell proliferation and cell differentiation.
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Construction of cis-Regulatory Input Functions of Yeast Promoters
Time of Update: 2021-02-14
Synthetic genetic constructs are well suited to precisely control and measure gene expression to constructcis -regulatory input functions.
These functions can be used to predict gene expression based on signal intensities transmitted to activators and repressors in the gene regulatory region.
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Reactor Configuration for Continuous Fermentation in Immobilized Systems: Application to Lactate Production
Time of Update: 2021-02-14
Results from bioreactor studies have demonstrated that immobilized cells have advantages over free cells, such as protection from toxic substances, increased plasmid stability and increased catalytic activity ( 1 ).
Because of these advantages, methods employing immobilized microbial cells are used extensively in many industrial applications.
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Applications of Interspersed Repeat Sequence Polymerase Chain Reaction
Time of Update: 2021-02-14
Analysis of complex genomes includes characterization of complete large-insert genomic libraries comprising several hundreds of thousands of clones.
Conventional methods to screen large-insert clone libraries for specific clones within a defined chromosomal interval are polymerase chain reaction ( PCR ) based using microsatellite markers.
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MAVisto: A Tool for Biological Network Motif Analysis
Time of Update: 2021-02-14
Data from high-throughput experimental methods are currently being used to construct complex biological networks.
The occurrence and frequency of certain motifs or pattern characterize the topology and often the functional space of a network.
