Beschreibung
The lipid-rich and otherwise challenging nature of many key tissues complicates many aspects of current research, and applications of the unique nature of lipoproteins and their biological effects has engendered unique and vital methodologies. In Lipoproteins and Cardiovascular Disease: Methods and Protocols, experts in the field present a compendium of advanced and classical molecular biology methods targeted towards lipoprotein, atherosclerosis, and vascular biology research, bringing together in a single volume an updated set of protocols and strategies for methods now driving the most recent advances, along with classical methods that are still widely used. Among the many topics covered in this cutting-edge work, the book delves into crucial techniques such as quantitative real-time PCR, microarrays, RT-PCR laser capture microdissection, and tissue-specific gene overexpression, knockout, and knockdown methodologies, including AAV as a liver-directed gene delivery vehicle. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective subjects, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and valuable notes which highlight tips on troubleshooting and avoiding known pitfalls. Comprehensive and easy to use, Lipoproteins and Cardiovascular Disease: Methods and Protocols serves both novices and experts alike as a complete guide for any researcher with an interest in lipoproteins and their significant biological effects.
Autorenporträt
InhaltsangabePart I: RNA and Gene Expression 1. Cloning Full-Length Transcripts and Transcript Variants Using 5' and 3' RACE Lita A. Freeman 2. Monitoring Gene Expression: Quantitative Real-Time RT-PCR Elke M. Wagner 3. Microarray Technology: Basic Methodology and Application in Clinical Research for Biomarker Discovery in Vascular Diseases Nalini Raghavachari 4. Northern Analysis of Gene Expression Lita A. Freeman 5. Laser Capture Microdissection for Analysis of Macrophage Gene Expression from Atherosclerotic Lesions Jonathan E. Feig and Edward A. Fisher Part II: Sequencing 6. Sequencing PCR-Amplified DNA in Lipoprotein and Cardiovascular Disease Research Victoria Youngblood and James G. Taylor VI 7. Introduction to Next-Generation Nucleic Acid Sequencing in Cardiovascular Disease Research Lena Diaw, Victoria Youngblood, and James G. Taylor VI Part III: Transgenic, Knockout, and Knockdown Methodologies 8. Strategies for Designing Transgenic DNA Constructs Chengyu Liu 9. Purification of Plasmid and BAC Transgenic DNA Constructs Chengyu Liu, Yubin Du, Wen Xie, and Changyun Gui 10. Pronuclear Microinjection and Oviduct Transfer Procedures for Transgenic Mouse Production Chengyu Liu, Wen Xie, Changyun Gui, and Yubin Du 11. Genotyping of Transgenic Animals by Real-Time Quantitative PCR with TaqMan Probes Boris Vaisman 12. Generation of General and Tissue-Specific Gene Knockout Mouse Models Xian-Cheng Jiang 13. Adeno-Associated Viruses as Liver-Directed Gene Delivery Vehicles: Focus on Lipoprotein Metabolism William R. Lagor, Julie C. Johnston, Martin Lock, Luk H. Vandenberghe, and Daniel J. Rader 14. Modulation of Lipoprotein Metabolism by Antisense Technology: Preclinical Drug Discovery Methodology Rosanne M. Crooke and Mark J. Graham Part IV: Special Topics 15. Chromatin Immunoprecipitation Grant D. Barish and Rajenda K. Tangirala 16. Measurement of Lecithin:Cholesterol Acyltransferase Activity with the Use of a Peptide-Proteoliposome Substrate Boris Vaisman and Alan T. Remaley 17. Native-Native 2D Gel Electrophoresis for HDL Subpopulation Analysis Lita A. Freeman 18. Western Blots Lita A. Freeman
