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Determination of the Gene Content of Salmonella Genomes by Microarray Analysis

Microarray technology provides a convenient and relatively inexpensive way of investigating the genetic content of bacterial genomes by comparative genomic hybridization. In this method, genomic DNA of an unknown bacterial strain of interest and that of a closely related sequenc ...

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Amplified Fragment Length Polymorphism Analysis of Salmonella enterica

Amplified fragment length polymorphism (AFLP) is a powerful PCR-based fingerprinting method and has the capacity to reveal variation around the whole genome by selectively amplifying a subset of restriction fragments for comparison. The restriction fragments analyzed are sma ...

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In Vivo Excision, Cloning, and Broad-Host-Range Transfer of Large Bacterial DNA Segments Using VEX-Capture

The performance of many bacterial genetic experiments would benefit from a convenient method to clone large sets of genes (20–100+kb) and transfer these genes to a wide range of other bacterial recipients. The VEX-Capture technique allows such large genomic segments to be cloned in vivo onto a b ...

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Salmonella Phages and ProphagesGenomics and Practical Aspects

Numerous bacteriophages specific to Salmonella have been isolated or identified as part of host genome sequencing projects. Phylogenetic analysis of the sequenced phages, based on related protein content using CoreGenes, reveals that these viruses fall into five groupings (P27- ...

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Handling the BSE Epidemic in Great Britain

Henry Ford thought that hlstory was bunk; Hegel is supposed to have said that “people and governments never have learnt anything from history,” and Disraeli advised us to “read no history.” The authors of this chapter are less dismissive, taking the alternative view that those who ignore history are ...

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Special Problems of Genetic Counseling in Adult-Onset Diseases: Huntingtons Disease as a Model

Neurological disorders impose a severe burden on affected individuals Inherited neurological disorders of late onset present their own specific difficulties in terms of genetic counseling, management, and patients’ psychological adaptation. Huntington’s disease (HD) is t ...

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Genotyping and Susceptibility of Sheep to Scrapie

Scrapie, as a disease of sheep and goats, has been recognized for over 300 yr and was first documented in England in 1730 (1). Sheep breeders petitioned the House of Commons of King George II in 1754 requesting the enforcing of regulations governing the sale and distribution of sheep This is believed to have ...

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Strain Typing Studies of Scrapie and BSE

The basis of strain vartation in scrapie and other transmissible spongrform encephalopathies is a crucial issue in the ongoing debate about the nature of the infectious agent. The clear evidence for the existence of multiple strains leads us to conclude that these agents carry some form of str ...

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PrP-Deficient Mice in the Study of Transmissible Spongiform Encephalopathies

The Transmissible Spongiform Encephalopathies (TSEs), such as scrapre, BSE, and Creutzfeldt-Jakob disease, are associated with alterations in the neural membrane protein or prion protein (PrP). This chapter will outline the gene targeting approaches that have been used to mutate the ...

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Transgenic Approaches to Prion Species-Barrier Effects

Like conventrinal viruses, prion isolates exhibit distinctive, and often restricted host-ranges. However, the molecular events that shape the hostranges of these two classes of pathogen are dissimilar, reflecting their fundamentally different life cycles. As discussed by Rid ...

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Methods for Studying Prion Protein (PrP) Metabolism and the Formation of Protease-Resistant PrP in Cell Culture and Cell-Free Systems

The pathogenesis of scrapie and other transmissible spongiform encephalopathies (TSEs) appears to be based on the posttranslational conversion of the host’s protease-sensitive prion protein (PrP-sen or PrPc) to abnormal protease-resistant forms (PrP-res or PrPSc). In vitro st ...

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Methods for Studying Prion Protein Amyloid

Prion encephalopathies are neurodegenerative diseases characterized by the accumulation of abnormal isoforms of the prion protein (PrP) and the deposition of PrP amyloid in the central nervous systein (CNS) (1, 2) The diseasespecific PrP molecules are distinguishable froin their ...

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Changing Methods for Discovering Antiviral Drugs

Viral diseases were largely untreatable 40 yr ago. Now effective and safe therapies are available. This has led to significant improvements in the quality of life for large numbers of patients. New viral diseases are, however, continuing to emerge and established viruses have been shown to dev ...

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Laboratory Safety Considerations

In the laboratory it is important that potentially pathogenic agents be controlled to protect the laboratory worker from infection and the experiment from contamination. The operation of a safe laboratory depends on many factors: the training and judgement of laboratory personnel; t ...

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Routine Quality Control Testing of Cell Cultures: Detection of Mycoplasma

The quality of cell cultures and their products is important in scientific research and is also critical where they are used for diagnosis and therapies. Fundamental aspects of quality of cell cultures are purity, correct identity, and freedom from adventitious agents as well as appropria ...

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Hep AD38 Assay: A High-Throughput, Cell-Based Screen for the Evaluation of Compounds Against Hepatitis B Virus

Approximately 5% of the world’s population has been infected with hepatitis B virus (HBV). Ten percent of these adults will become chronic carriers, as will 95% of the infants infected perinatally. Those that do become chronically infected with HBV are at increased risk of developing liver dysf ...

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Hepatitis B Virus Cell Culture Assays for Antiviral Activity

Hepatitis B Virus (HBV) is a causative agent of both acute and chronic hepatitis, a major etiologic factor of primary hepatocellular carcinoma, and a serious global health problem, with over 240 million estimated chronically infected individuals. Although there are several promising ...

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A Rapid Microtiter Assay for Duck Hepatitis Virus Reverse Transcriptase

The duck hepatitis B virus (DHBV) genome contains a pol gene that codes for the viral polymerase protein. This enzyme, which is essential for the replication of the virus, has multiple activities including an RNA directed DNA polymerase or reverse transcriptase (RT) activity, an RNase H activit ...

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Assays for Measuring Hepadnaviral Supercoiled DNA

Amajor reason for treatment failure during antiviral therapy of chronic hepatitis B infection is thought to be the persistence of the key replicative intermediate, the viral covalently closed circular (CCC) or supercoiled DNA (1,2). Investigators studying the structure and functi ...

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Assays for Hepatitis B Virus DNA-and RNA-Dependent DNA Polymerase Activities

Genomes of the hepatitis B viruses (HBVs) consist of approx 3.2 kb of partly double-stranded DNA containing three or four overlapping open reading frames, the largest of which encodes the viral polymerase (Pol) protein. After entry into the cell and uncoating, the viral genome is transported to t ...

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