Vascular Disease: Molecular Biology and Gene Transfer Protocols

ISBN-13: 9780896037311 / Angielski / Twarda / 1999 / 441 str.

Andrew H. Baker

Opis

Molecular biology has revolutionized research into vascular disease. Over the past 20 years molecular techniques have enabled us to both elucidate - lecular mechanisms in vascular disease and identify appropriate therapies. The vast explosion in technical knowledge and the array of protocols that become more advanced and intricate by the day lead us into new and exciting areas of research that were previously unobtainable. Vascular Disease: Molecular Biology and Gene Transfer Protocols - scribes today's most powerful molecular methods for the investigation of the pathogenesis of vascular disease. The protocols are highly detailed, allowing beginners who have little experience in either vascular biology or molecular biology to embark on new molecular projects. This book is also suited to more experienced molecular biologists who wish to grasp new methods for stu- ing the involvement of genes in normal vascular physiology and in diseased states. It is well established that cardiovascular disease progression has a s- stantial genetic influence. Part I describes three methods that have been used successfully to identify specific mutations in candidate genes involved in c- diovascular disorders. These mutations include both single-stranded conf- mational polymorphism analysis and heteroduplex detection methods. In addition, technology to map new genes to specific regions of chromosomes by high-resolution mapping is described.

Recenzje

"...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

Spis treści

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 High-Efficien

Blurb

In Vascular Disease: Molecular Biology and Gene Therapy Protocols, 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. Accurate methods to quantify apoptosis in both cultured cells and pathological specimens are also given. 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. Powerful and productive, the techniques presented here operate across a wide range of exciting research areas, and promise spectacular therapeutic breakthroughs in the ongoing battle against vascular disease.