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The aim of this book is to provide detailed protocols for studying the
molecular biology of the pathogen Mycobacterium tuberculosis, and its interactions
with host cells. As established mycobacterial laboratories move towards
exploiting the genome, and laboratories with expertise in other fields
apply them to mycobacteria, both traditional and novel methodologies need to
be reviewed. Thus the chapters in Mycobacterium tuberculosis Protocols range
from perspectives on storage of strains and safety issues to the application of
the latest functional genomics technologies.
The last few years have been remarkable ones for research into M. tuberculosis.
The most important landmark by far has been the completion of the
genome sequence of the widely studied H37Rv strain (1). We can now predict
every protein and RNA molecule made by the pathogen. This information is or
will soon be enriched by the addition of genome sequences of other strains
from the M. tuberculosis complex: a second strain of M. tuberculosis, Mycobacterium
bovis, and the vaccine strain, M. bovis BCG. Valuable comparative
data will also be provided by the genome sequences of Mycobacterium leprae,
Mycobacterium avium, and Streptomyces coelicolor. Another recent milestone
for M. tuberculosis has been the development of efficient mutagenesis methodologies,
the lack of which has been a major handicap in functional studies.
The new challenges to researchers are first to use this information and these
techniques in combination with the battery of methodologies being developed
around the world to exploit all genome data, so-called functional genomics
research, which includes transcriptomics and proteomics. The second challenge
is to integrate them with other disciplines of active research such as
immunology, cell biology, and biochemistry. Mycobacterium tuberculosis
Protocols incorporates both of these aspects in the methods described.
This book is aimed at people who are actively working on M. tuberculosis.
However, there is much that will be relevant to work on other mycobacteria
and on such phylogenetically related organisms as corynebacteria and streptomycetes.
It is intended both for people with experience in handling M. tuberculosis
and those who are new to the field.
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