Vascular Disease: Molecular Biology and Gene Transfer Protocols

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August 1999



Andrew Baker and a noted panel of expert investigators describe today's most powerful molecular methods for investigating the pathogenesis of vascular disease. These detailed, easy-to-follow techniques range from methods that have been used successfully to identify specific mutations involved in cardiovascular disorders, to those for transferring genes associated with cardiovascular disease into various vascular cell types by in vitro and in vivo routes. There are methods to identify novel genes and generate full-length cDNAs, to study gene transcription and promoter activity easily and effectively, and to ascertain precisely gene expression levels within the individual cell types in different pathophysiological conditions. Vascular Disease: Molecular Biology and Gene Therapy Protocols offers today's vascular biologist and gene therapist an unprecedented ability to study the pathogenesis of vascular disease and readily to probe the potential for gene-based therapies.


Part I. Genetics in Vascular Disease

Detection of Mutations and DNA Polymorphisms in Genes Involved in Cardiovascular Diseases by Polymerase Chain Reaction-Single-Strand Conformation Polymorphism Analysis
Shu Ye and Adriano M. Henney

Analysis of Genetic Variants in Cardiovascular Risk Genes by Heteroduplex Analysis
Ana Cenarro, Fernando Civeira, and Miguel Pocovi

Radiation Hybrid (RH) Mapping of Human Smooth Muscle-Restricted Genes
Joseph M. Miano, Emilio Garcia, and Ralf Krahe

Part II. Isolation of Genes Expressed in Vascular Tissue

Efficient Extraction of RNA from Vascular Tissue
Catherine F. Townsend, Christopher M. H. Newman, and Sheila E. Francis

Preparation of cDNA Libraries from Vascular Cells
Mark E. Lieb and Mark B. Taubman

Screening cDNA Libraries Using Partial Probes to Isolate Full-Length cDNAs from Vascular Cells
Csilla Csortos, Virginie Lazar, and Joe G. N. Garcia

Cloning Full-Length cDNAs from Vascular Tissues and Cells by Rapid Amplification of cDNA Ends (RACE) and RT-PCR
Rong-Fong Shen

Use of cDNA Representational Difference Analysis to Identify Disease-Specific Genes in Human Atherosclerotic Plaques
Kerry Tyson and Catherine Shanahan

The Use of Differential mRNA Display (DDRT-PCR) to Identify Genes Differentially Expressed in Normal and Diseased Vascular Cells
Paul J. Adam

Identification of Novel Protein Kinases in Vascular Cells
Ian Zachary, Spiros Servos, and Barbara Herren

Part III. Methods for Mapping Transcriptional Start Sites and Measurement of Promoter Activity in Vascular Cells

Primer Extension Analysis to Map Transcription Start Sites of Vascular Genes
Yutaka Kitami and Kunio Hiwada

Use of Liposome-Mediated DNA Transfection to Determine Promoter Activity in Smooth Muscle Cells
Rosalind P. Fabunmi

Nuclear Run-On Assayto Study Gene Transcription in Vascular Cells
Ulrich Laufs and James K. Liao

Electromobility Shift Analysis (EMSA) Applied to the Study of NF-kappa B Binding Interactions in Vascular Smooth Muscle Cells
Todd Bourcier

Part IV. Molecular Analysis of mRNA Expression in Vascular Cells

Measurement of Gene Expression in the Vascular Wall by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) Analysis
Joachim Fruebis

Northern Blot Analysis to Quantify Gene Expression in Vascular Diseases
Mingyi Chen and Tatsuya Sawamura

Localization of Gene Expression in Human Atherosclerotic Lesions by In Situ Hybridization
Lee D. K. Buttery and Julia M. Polak

RNase Protection Assays for Quantification of Gene Expression in Vascular Tissue
Nengyu Yang, Shuli Wang, and James E. Faber

Part V. Molecular Methods to Study Apoptosis and Phenotypic Changes in Vascular Cells and Tissue

Detection of Apoptosis in Atherosclerosis and Restenosis by Terminal dUTP Nick End Labeling (TUNEL)
Mark Kockx, Johannes Muhring, and Michiel Knaapen

In Vitro Detection of Apoptosis in Isolated Vascular Cells
Shiu-Wan Chan and Martin R. Bennett

Molecular Assessment of the Phenotypic Changes Associated with Smooth Muscle Cells Using Two-Dimension Electrophoresis and Microsequence Analysis
Rachel Johnatty, Giulio Gabbiani, and Pascal Neuville

Part VI. In Vitro Methods to Express Foreign Genes in Vascular Cells

Simple Methods for Preparing Recombinant Adenoviruses for High-Efficiency Transduction of Vascular Cells
Stuart A. Nicklin and Andrew H. Baker

Generation of Recombinant Adeno-Associated Viruses for Delivery of Genes into Vascular Cells
Carmel M. Lynch

Hemagglutinating Virus of Japan Liposome-Mediated Gene Delivery to Vascular Cells
Yoshikazu Yonemitsu and Yasufumi Kaneda



"...The chapters are clearly written so that complex techniques are reduced to their individual components...this book will be a major asset to a molecular biology laboratory involved in vascular research."-Atherosclerosis
"This collection of very detailed molecular biology protocols was compiled...The first five parts present protocols that are designed to identity and characterize genes in cardiovascular disease, including methods for detecting specific gene mutation, identifying new genes, characterizing the levels and regulation of expression of the new genes, and apoptotic methods. The final two parts contain in vivo and in culture gene transfer protocols used in cardiovascular disease and protocols for clinical gene therapy....an excellent reference for molecular biologists in the field of vascular disease." - Quarterly Review of Biology
"...the authors of the individual chapters have made every effort to relate their particular techniques to the study of vascular tissues....the editor has assembled a broad base of well established authors to describe their protocols....this is a good book....this book may prove worthwhile in helping to ease the load on the typical physician-scientist trying to care for patients and at the same time direct his or her laboratory research program."- Texas Heart Institute Journal
"The book is the result of the contribution of distinguished experts in the field. Each chapter of the book is introduced by a theoretical summary of the goal of the technique with excellent diagrams followed by the material used, the basic procedures and the methods. Critical notes and references end each chapter. This important publication directed to molecular biologists covers the latest aspects of molecular procedures for the study of cardiovascular diseases and confirms, if necessary, the irreversible revolution in cardiovascular research."- Acta Cardiologica
EAN: 9780896037311
ISBN: 0896037312
Untertitel: Molecular Biology and Gene Transfer Protocols. 'Methods in Molecular Medicine'. 1999. Auflage. Sprache: Englisch.
Erscheinungsdatum: August 1999
Seitenanzahl: 441 Seiten
Format: gebunden
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