实验基础

The protozoan Trypanosoma brucei is the most destructive parasite of domestic livestock in sub-Saharan Africa. The parasite lives extracellularly in its two hosts; alternately in the bloodstream of a mammal, and in the midgut and subsequently the salivary glands of the tsetse fly (Glossi ...

Any biochemical analysis is usually made easier by the availability of large numbers of cells to be analyzed, and one of the reasons for the position held by Trypanusoma brucei as the best characterized parasite is the relative ease with which it can be cultured in the laboratory. The ability to culture ...

The scope of this book does not allow a complete description of the many techniques available for purification and treatment of reagents for facilitating immunoassays in general. There is a large amount of literature covering techniques, and this can be consulted for specific problems. The ...

In this chapter, the use of control charts to both continuously evaluate testing in individual laboratories as well as provide data for external monitoring is examined in detail. The data is based on the publication by D. E. Rebeski' et al., “Charting methods to monitor the operational performance ...

This chapter deals with control charts to monitor the performance of Indirect ELISAs. An Indirect ELISA kit for the detection of antibodies against Brucella is used to demonstrate the methods. Many of the features explained in Chapter 9 are relevant to this chapter; some repetition is intended, ...

Standard protocols for definite species identification of nontuberculous mycobacteria by phenotypic characteristics include complex and time-consuming procedures. Other methods based on lipid analysis, such as high-performance liquid chromatography (HPLC), thin ...

Tuberculosts (TB) IS the most sign&ant smgle mfecttous cause of morbrdtty and mortalrty, producing eight million new cases and killing three million people annually. It causes approximately one quarter of all avoidable adult deaths from mfectton (1–3).

Mycobacteria, like other bacteria, respond to environmental signals with changes in gene expression regulated at the level of transcription. The significance of gene-expression patterns in pathogens, such asMycobacterium tuberculosis, is that specific signals encounter ...

Mycobacterium bovis Bacille Calmette Gu�rin (BCG) has several features that make it desirable as a vaccine vehicle for foreign antigens. BCG is a live vaccine and stimulates a strong cell-mediated immune response, is extremely safe, and can be given at birth (1). Using recently developed tools to ...

Over the past 10 years, major advances have been made in the field of molecular mycobacterioiogy. These include the generation of shuttle vectors for use in both Escherichia coli and mycobacteria, electroporation protocols to introduce these plasmids into mycobacteria and promoter c ...

Bioluminescence is the product of two distinct enzymes, firefly luciferase and bacterial luciferase. The application of the firefly enzyme in the study of mycobacteria has been described by other groups (1,2) (see Chapter 31). The bacterial-luciferase enzyme is a drmer of approx 80 kDa, cons ...

Gene replacement is a powerful technique that exploits the innate ability of an organism to recombme homologous regions of DNA. It allows for the specrfic replacement of targeted genomic sequences with copies of those sequences carrying defined mutations, and therefore can facilitate ...

Much of the present knowledge of the bacterial cell emanates from studies of spontaneous mutants and mutants obtained as a result of random or directed mutagenesis. Spontaneous mutants in a specific gene can generally be Isolated only if the mutation leads to a selectable phenotype, e.g., muta ...

The process of homologous recombination IS essential to all living organisms. It is important for the generation of genetic diversity, the maintenance of genomic integrity, and the proper segregation of chromosomes. At least 25 gene products (RecA, helicases, nucleases, ligases, sing ...

Mobile generic elements are discrete DNA fragments encoding their own transposltlon functions, which enable them to move from one site in the DNA to another, independently of host-recombination functions (such as RecA). Recombination by transposition occurs without homology bet ...

The creation and characterization of mutations within mycobacterial genomes is crucial for the analysis of gene function and regulation in members of the species. Such mutants could be used to elucidate mechanisms of virulence and survival of pathogenic mycobacteria such as Mycobac ...

Mycobacteriophages are viruses that infect the mycobacteria Historically, they have been used to overcome major road blocks in the genetic manipulation of mycobacteria. Mycobacteriophages have been used to demonstrate mycobacterlal transfection (1) and transduction (2). A ...

The study of mycobacterial genomes has exploded during the last 10 yr. Initially, no systems were available for the du-ect manipulation of mycobacterial genes in mycobactena, so Escherichia coli was used as the primary cloning host. Several genomic libraries were created (1–5) in E. coli. Alth ...

Bacterial conjugation is a phenomenon that is more widespread than was originally thought. It medtates genetrc exchange not only between cells of the same species, but also between a wide variety of unrelated organisms. Conjugation between gram-negative and gram-positive bacteria w ...

A significant proportion of the world’s population is infected with Mycobacterium tuberculoszs, with estimates in 1992 of 8 million new mfections and 3 million deaths from tuberculosis (TB) per annum (1). There are several other causes for concern. There IS a close assoclatlon between the hum ...