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Extraction and Purification of Nucleic Acids from Schistosomes
Time of Update: 2021-02-17
Once an extraction procedure has begun, it is advisable to proceed as rapidly as possible to its conclusion or the nucleic acids may degrade.
As a general rule, all procedures should be performed at 0–4�C unless otherwise mentioned.
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Expression of Genes in Mycobacteria
Time of Update: 2021-02-17
Similarly, the ability to overexpress cloned genes in bacteria has been important in revealing the functions of a great diversity of heterologous proteins.
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Molecular Methods to Study Transcriptional Regulation of Clostridium difficile Toxin Genes
Time of Update: 2021-02-17
TcdR is an alternative RNA polymerase sigma factor that positively regulates toxin gene transcription as well as its own.
TcdC negatively regulates toxin synthesis by interfering with the RNA polymerase formed with TcdR.
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Bacterial Identification and Subtyping Using DNA Microarray and DNA Sequencing
Time of Update: 2021-02-17
In this chapter, we will review (1) the use of DNA microarray using fluorescence and infrared imaging detection for identification of pathogenic bacteria, and (2) use of pyrosequencing in DNA cluster analysis to fingerprint bacterial phylogenetic trees.
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Ribozyme-Mediated Downregulation of Gene Expression in Transgenic Mice
Time of Update: 2021-02-17
Transcript-specific downregulation of RNA levels and activity using antisense RNA techniques represents a promising approach toward this end.
One major obstacle is the need to maintain relatively high intracellular levels of the antisense RNA, which interacts stoichiometrically with the target transcript.
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Safe Dispatch and Transport of Biological Material
Time of Update: 2021-02-17
The demands on the export, packaging, and shipping of biological material are manifold and the number of possible mistakes is endless.
The regulations for shipping biological substances by postal mail are laid down by the Universal Postal Union (UPU).
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Production of Heterologous Proteins in Yeast With the Aid of the Hsp150 Carrier
Time of Update: 2021-02-17
Biologically active mammalian glycoproteins can be produced inSaccharomyces cerevisiae andPichia pastoris by joining them to a fragment of a natural secretory glycoprotein ofS.
Here we describe the use of the Hsp150Δ carrier inP.
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Identification of Novel Pathogenicity Genes by PCR Signature-Tagged Mutagenesis and Related Technologies
Time of Update: 2021-02-17
This is a PCR-based STM mutation-based screening method using a population of bacterial mutants for the simultaneous identification of multiple virulence genes in microbial pathogens by negative selection.
PCR-based STM represent a genomics-based method for in vivo high-throughput screening of new virulence factors.
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Enumeration of Bacteriophages Using Flow Cytometry
Time of Update: 2021-02-17
The flow cytometric method allows the detection and discrimination of a wide variety of viruses of different morphology, genome type, and size.
The present paper describes an optimized protocol for the enumeration of bacteriophages using a standard benchtop flow cytometer.
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Serological Identification of Mycoplasmas by Growth and Metabolism Inhibition Tests
Time of Update: 2021-02-17
In the diagnostic laboratory, unknown mycoplasma isolates are inoculated onto solid media and examined for growth inhibition in the presence of antiserum raised against known mycoplasma species.
Zones of inhibition of >2 mm are usually considered significant evidence that the isolate belongs to the species represented by the inhibitory antiserum.
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Expression and Purification of Recombinant Histones and Nucleosome Reconstitution
Time of Update: 2021-02-17
The extent of heterogeneity and modification strongly depend on the type and developmental state of the tissue from which chromatin is isolated and can vary significantly between different batches.
Fourth, and most important, only naturally occurring histone proteins can be obtained by this method.
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Immunomagnetic Separation of Pathogenic Organisms From Environmental Matrices
Time of Update: 2021-02-17
One of the most difficult challenges in the analysis of environmental samples is to separate the organism of interest from a sample that is high in background debris. Immunomagnetic separation (IMS)
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Sampling Water Bodies: Tangential Flow Filtration
Time of Update: 2021-02-17
The study of microbial community structure requires a representative sample of that community for processing in a form that is free from contaminants that will interfere with the analysis.
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The Relationship between Helicobacter Pylori Infection and Remnant Gastritis in 152 Cases
Time of Update: 2021-02-17
After partial gastric excision, 60% to 100% of patients developed residual gastritis, and the rate of HP-positive detection in the residual stomach was reported at 25% to 71%.
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Ins and Outs of Mycobacterial Plasmids
Time of Update: 2021-02-17
The importance of plasmids for molecular research cannot be underestimated.
Fortunately, today the mycobacterial research community has a number of these genetic tools at its disposal, and the development of these tools has greatly accelerated the study of mycobacterial pathogens.
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Biological and Clinical Basis for Molecular Studies of Interferons
Time of Update: 2021-02-17
The cytokine family of interferons (IFNs) has multiple functions, including antiviral, antitumor, and immunomodulatory effects and regulation of cell differentiation.
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Clostridium difficile: No Longer an Enigmatic Pathogen
Time of Update: 2021-02-17
Coupled with the increased interest in this organism researchers now have a greater understanding of its genetic content and molecular epidemiology; a direct consequence of the multipleC.
Concurrent with the sequencing efforts have been the development of tools to genetically manipulate the organism.
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Quantitation and Structural Analysis of SV40 RNAs
Time of Update: 2021-02-17
These proteins play roles in replication of the viral genome and transcriptional transactivation of the late genes (see refs.
Thecis -acting transcriptional elements that regulate the synthesis of both the early and late viral RNAs (see refs.
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Analysis of Human Cytomegalovirus Using the Polymerase Chain Reaction
Time of Update: 2021-02-17
Similarly, reverse transcription-PCR ( RT-PCR ) allows the study of targeted gene expression, by reverse transcription of RNA to complementary DNA ( cDNA ), followed by amplification of target DNA using predetermined primers.
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Production of Baculovirus-Expressed Recombinant Proteins in Wave Bioreactors
Time of Update: 2021-02-17
Wave Bioreactors are relatively new to the field of protein production from insect cells infected with recombinant baculoviruses.
Excellent insect cell growth and production of proteins from many classes can be achieved with these units.
