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Dictyostelium discoideum is a well-establish eukaryotic model
organism that offers unique advantages for studying fundamental
cellular processes, including signal transduction, random and
directed cell motility, cytokinesis, endocytosis and vesicle
transport and development. Dictyostelium is also increasingly used
for the investigation of human disease genes and the crosstalk
between host and pathogen. The availability of the genome sequence
at a mouse click together with a whole range of supporting
information and resources along with a powerful armoury of
molecular genetics techniques have considerably enhanced the
experimental attractiveness of D. discoideum in recent years. The
second edition of Dictyostelium discoideum Protocols incorporates
the most recent developments in a number of fields. The book is
divided into four parts. The first part provides an introduction to
the amoebozoa and community resources. The second part presents
large-scale analysis methods made possible by the completion of the
Dictyostelium genome sequence. The third part is dedicated to
molecular genetics techniques, cell biological, biochemical and
biophysical methods. The chapters in the fourth part describe the
use of Dictyostelium as a model system to study vesicle formation,
trafficking and infection by bacterial pathogens. Written in the
highly successful Methods in Molecular Biology (TM) series format,
chapters include introductions to their respective topics, lists of
the necessary materials and reagents, step-by-step, readily
reproducible laboratory protocols, and key tips on troubleshooting
and avoiding known pitfalls. Authoritative and practical,
Dictyostelium discoideum Protocols, Second Edition seeks to aid
scientists in the application of innovating techniques to study a
range of fundamental biological processes in this attractive model
organism.
This detailed book expands upon the previous edition with a
collection of methods for those performing experimental work on
small GTPases of the Rho family. Split into four sections, the
volume explores computational modeling and imaging procedures,
biochemical methods related to post-translational modifications of
Rho GTPases as well as some high throughput methods, functional
assays that allow for monitoring the consequences of manipulating
Rho GTPases in a variety of cell types and cell biology processes,
and techniques specifically designed for studies in selected
non-mammalian model organisms (zebrafish, social ameba, plants and
algae). Written for the highly successful Methods in Molecular
Biology series, chapters include introductions to their respective
topics, lists of the necessary materials and reagents,
step-by-step, readily reproducible laboratory protocols, and tips
on trouble shooting and avoiding known pitfalls. Authoritative and
up-to-date, Rho GTPases: Methods and Protocols, Second Edition
constitutes an invaluable tool for all those with an interest in
this remarkable family of signaling proteins.
Although initially described as major regulators of cytoskeletal
remodeling, Rho GTPases have been implicated in the establishment
of polarity, endocytosis, vesicle trafficking, morphogenesis,
cytokinesis, transcriptional activation, cell cycle progression,
and apoptosis, to mention a few. In addition, Rho GTPases have
acquired medical relevance because of their participation in
tumorigenesis and metastasis, in cardiovascular conditions, and as
targets of infectious agents. The field has broadened even more
with the contribution of studies in model organisms (plants,
amoebas, fungi, invertebrates), each adding a particular view to
the complexity of the family and vastly enriching our perception of
these important signaling components. Divided into five convenient
sections, Rho GTPase: Methods and Protocols provides an historical
overview of the field and an account of the phylogenetics of the
Rho family, general biochemical methods, and functional assays that
allow monitoring the consequences of manipulating Rho GTPases in a
variety of contexts. Additionally, the volume devotes a section to
advanced imaging methods and to recently developed high throughput
methods, closing with techniques specifically designed for studies
in selected non-mammalian model organisms. Written in the
successful Methods in Molecular Biology (TM) series format,
chapters include introductions to their respective topics, lists of
the necessary materials and reagents, step-by-step, readily
reproducible protocols, and notes on troubleshooting and avoiding
known pitfalls. Authoritative and easily accessible, Rho GTPase:
Methods and Protocols provides techniques that are standard for
researchers in the field but also includes protocols for those who,
being already familiar with some of the techniques, wish to explore
additional aspects.
Dictyostelium discoideum is a well-establish eukaryotic model
organism that offers unique advantages for studying fundamental
cellular processes, including signal transduction, random and
directed cell motility, cytokinesis, endocytosis and vesicle
transport and development. Dictyostelium is also increasingly used
for the investigation of human disease genes and the crosstalk
between host and pathogen. The availability of the genome sequence
at a mouse click together with a whole range of supporting
information and resources along with a powerful armoury of
molecular genetics techniques have considerably enhanced the
experimental attractiveness of D. discoideum in recent years. The
second edition of Dictyostelium discoideum Protocols incorporates
the most recent developments in a number of fields. The book is
divided into four parts. The first part provides an introduction to
the amoebozoa and community resources. The second part presents
large-scale analysis methods made possible by the completion of the
Dictyostelium genome sequence. The third part is dedicated to
molecular genetics techniques, cell biological, biochemical and
biophysical methods. The chapters in the fourth part describe the
use of Dictyostelium as a model system to study vesicle formation,
trafficking and infection by bacterial pathogens. Written in the
highly successful Methods in Molecular Biology (TM) series format,
chapters include introductions to their respective topics, lists of
the necessary materials and reagents, step-by-step, readily
reproducible laboratory protocols, and key tips on troubleshooting
and avoiding known pitfalls. Authoritative and practical,
Dictyostelium discoideum Protocols, Second Edition seeks to aid
scientists in the application of innovating techniques to study a
range of fundamental biological processes in this attractive model
organism.
Although initially described as major regulators of cytoskeletal
remodeling, Rho GTPases have been implicated in the establishment
of polarity, endocytosis, vesicle trafficking, morphogenesis,
cytokinesis, transcriptional activation, cell cycle progression,
and apoptosis, to mention a few. In addition, Rho GTPases have
acquired medical relevance because of their participation in
tumorigenesis and metastasis, in cardiovascular conditions, and as
targets of infectious agents. The field has broadened even more
with the contribution of studies in model organisms (plants,
amoebas, fungi, invertebrates), each adding a particular view to
the complexity of the family and vastly enriching our perception of
these important signaling components. Divided into five convenient
sections, Rho GTPase: Methods and Protocols provides an historical
overview of the field and an account of the phylogenetics of the
Rho family, general biochemical methods, and functional assays that
allow monitoring the consequences of manipulating Rho GTPases in a
variety of contexts. Additionally, the volume devotes a section to
advanced imaging methods and to recently developed high throughput
methods, closing with techniques specifically designed for studies
in selected non-mammalian model organisms. Written in the
successful Methods in Molecular Biology (TM) series format,
chapters include introductions to their respective topics, lists of
the necessary materials and reagents, step-by-step, readily
reproducible protocols, and notes on troubleshooting and avoiding
known pitfalls. Authoritative and easily accessible, Rho GTPase:
Methods and Protocols provides techniques that are standard for
researchers in the field but also includes protocols for those who,
being already familiar with some of the techniques, wish to explore
additional aspects.
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