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Main description:
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.
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
High-Efficien
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 456
Weight: 735g
Availability: Available
Subcategories: Vascular
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