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Cystic Fibrosis Methods and Protocols

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Titel: Cystic Fibrosis Methods and Protocols

ISBN: 0896038971
EAN: 9780896038974
Auflage 2002.
XVI, 615 p.
Sprache: Englisch.
Herausgegeben von William R. Skach

28. Februar 2002 - gebunden - 615 Seiten

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Since the cloning of the cystic fibrosis transmembrane conductance re- lator (CFTR) nearly a decade ago, cystic fibrosis (CF) research has witnessed a dramatic expansion into new scientific areas. Basic researchers, clinicians, and patients increasingly rely on fundamental techniques of genetics, molecular biology, electrophysiology, biochemistry, cell biology, microbiology, and immunology to understand the molecular basis of this complex disease. Research into the pathophysiology of CF has established numerous paradigms of ion channel dysfunction that extend from inflammation and infection in the airways of patients to basic mechanisms of protein processing and regulation in intracellular components. With these rapid advances has come an increasing need for research scientists to understand and utilize a growing array of basic laboratory tools. This volume of Methods in Molecular Medicine, Cystic Fibrosis Methods and Protocols satisfies that need by providing detailed protocols for the laboratory techniques used throughout CF research. From electrophysiology and cell biology, to animal models and gene therapy, the comprehensive set of methods covered here provide step-by-step instructions needed for investigators to incorporate new approaches into their research programs. Contributions have been chosen to reflect the rich diversity of techniques and to provide a cohesive framework for understanding challenges that are currently at the forefront of CF research. It is hoped that this volume will serve as a valuable reference that will not only foster interdisciplinary investigations into current problems encountered in CF, but also facilitate the translation of new scientific discoveries into clinical solutions.
Part I. Genetics of Cystic Fibrosis

CFTR Mutation Detection by Multiplex Heteroduplex (mHET) Analysis on MDE Gel
Julian Zielenski, Isabel Aznarez, Tuncer Onay, John Tzounzouris, Danuta Markiewicz, and Lap-Chee Tsui

cDNA Microarrays for Pharmacogenomic Analysis of Cystic Fibrosis, Meera Srivastava
Ofer Eidelman, and Harvey B. Pollard

Natural Animal Models of Human Genetic Diseases
Jeffrey J. Wine, Michael Dean, and Damjan Glavac

Part II. CFTR Structure and Function: Structure, Gating, and Regulation

Electrophysiological Approach to Studying CFTR
Horst Fischer

Quantitative Analysis of ATP-Dependent Gating of CFTR
Allan Powe, Zhen Zhou, Tzyh-Chang Hwang, and Georg Nagel

CFTR Regulation by Phosphorylation
Tang Zhu, Deborah A. R. Hinkson, David Dahan, Alexandra Evagelidis, and John W. Hanrahan

Transepithelial Measurements of Bicarbonate Secretion in Calu-3 Cells
Robert J. Bridges

Transepithelial Impedance Analysis of Chloride Secretion
Ashvani K. Singh, Sangeeta Singh, Daniel C. Devor, Raymond A. Frizzell, Willy van Driessche, and Robert J. Bridges

Studies of the Molecular Basis for Cystic Fibrosis Using Purified Reconstituted CFTR Protein
Ilana Kogan, Mohabir Ramjeesingh, Canhui Li, and Christine E. Bear

Probing CFTR Channel Structure and Function Using the Substituted-Cysteine-Accessibility Method
Myles H. Akabas

Methods for the Study of Intermolecular and Intramolecular Interactions Regulating CFTR Function
Anjaparavanda P. Naren

Fluorescent Indicator Methods to Assay Functional CFTR Expression in Cells
Alan S. Verkman and Sujatha Jayaraman

Expression, Folding, and Degradation

Immunolocalization of CFTR in Intact Tissue and Cultured Cells
Christopher R. Marino

Analysis of CFTR Trafficking and Polarization Using Green Fluorescent Protein and Confocal Microscopy
Bryan D. Moyer and Bruce A. Stanton

CFTR Folding and Maturation in Cells
Mohamed Benharouga, Manu Sharma, and Gergely L. Lukacs

Isolation of CFTR: Chaperone Complexes by Co-Immunoprecipitation
Geoffrey C. Meacham and Douglas M. Cyr

CFTR Expression and ER-Associated Degradation in Yeast
Yimao Zhang, Susan Michaelis, and Jeffrey L. Brodsky

Manipulating the Folding Pathway of -F508 CFTR Using Chemical Chaperones
Marybeth Howard and William J. Welch

CFTR Degradation and Aggregation
Michael J. Corboy, Philip J. Thomas, and W. Christian Wigley

In Vitro Reconstitution of CFTR Biogenesis and Degradation
Jon Oberdorf and William R. Skach

In Vitro CFTR Folding Assays
Rhesa D. Stidham, W. Christian Wigley, and Philip J. Thomas

Analysis of CFTR Endocytosis by Cell Surface Biotinylation
Kelly Weixel and Neil A. Bradbury

Regulatory Complexes

CFTR Regulation of ENaC
Scott H. Donaldson, Elaine G. Poligone, and M. Jackson Stutts

Yeast Two-Hybrid Identification and Analysis of Protein Interactions with CFTR
Viswanathan Raghuram, Kenneth R. Hallows, and J. Kevin Foskett

Biochemical Assays for Studying Indirect Interactions Between CFTR and the Cytoskeleton
Peter J. Mohler, Patricia L. Kultgen, M. Jackson Stutts, and Sharon L. Milgram

CFTR-Associated ATP Transport and Release
Marie E. Egan

Part III. Pathophysiology of Cystic Fibrosis

Inflammatory Mediators in CF Patients,
Jay B. Hilliard, Michael W. Konstan, and Pamela B. Davis

Bacterial Colonization and Infection in the CF Lung
Scott D. Sagel, Elaine B. Dowell, and Frank J. Accurso

Antimicrobial Peptides and Proteins in the CF Airway,
Alexander M. Cole and Tomas Ganz

Bacterial-Epithelial Interactions
Ruth Bryan and Alice Prince
"The coverage of methodologies is extremely broad and will be useful for those wanting to find out more about some of the less routine techniques used in the analysis of the CFTR protein/chloride channel. The style of protocol-based chapters enables in manuscripts due to considerations of space or potential repetition of previously published techniques. As such many of the chapters are valuable." -Molecular Biotechnology

"Undoubtedly this book will be regarded as essential reading for scientists in the field. It provides details of research methodology, which are rarely published in peer reviewed journals. It will also enable researchers to identify subtle differences in techniques in their own laboratories from those described in these chapters. As such, it will be a very important book for all those who conduct laboratory-based research into cystic fibrosis." -Journal of Paediatrics and Child Health
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