Pub. Date:
Springer-Verlag New York, LLC
Vascular Disease: Molecular Biology and Gene Transfer Protocols / Edition 1

Vascular Disease: Molecular Biology and Gene Transfer Protocols / Edition 1

by Andrew H. Baker
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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.

Product Details

ISBN-13: 9781617371554
Publisher: Springer-Verlag New York, LLC
Publication date: 11/09/2010
Series: Methods in Molecular Medicine , #30
Edition description: Softcover reprint of hardcover 1st ed. 1999
Pages: 441
Product dimensions: 5.98(w) x 9.02(h) x 0.04(d)

Table of Contents

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


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