E-Book Overview
Jose Ordovas compiles a cutting-edge collection of molecular protein techniques for studying advanced aspects of lipoprotein structure and metabolism. These step-by-step methods allow both the beginning and the advanced researcher to carry out not only fundamental, but also the most advanced genetic screening techniques, including RT-PCR and the preparation of relevant models of transgenic and knockout mice. In addition, novel techniques are presented to characterize plasma lipoproteins, their subfractions, and their modifications, and to determine the activity of relevant enzymes and carrier proteins. Each method is described in easily reproducible detail by a hands-on expert who has perfected it. Lipoprotein Protocols offers many powerful tools for unraveling the complexity of lipoprotein metabolism and determining the individual risk of coronary heart disease.
E-Book Content
1 PCR Methodology Applied to Genetic Studies of Lipoprotein Metabolism and Atherosclerosis Shelley A. Cole and James E. Hixson
1. Introduction During the 1980s molecular genetics became widely applied to hpoprotein research. The cloning of candrdate genes for atherosclerosis (apohpoprotems, lipid-processing enzymes, lrpid transfer proteins, and hpoprotem receptors) allowed the identrficatron of mutattons responstble for rare dysltpoproteinemtas, as well as more common genettc variants. The cloned candidate genes were used as probes on Southern blots to find polymorphtc nucleottde substituttons at restriction enzyme sates(restrtction fragment-length polymorphisms or RFLPs) that could be used as genetic markers m linkage and assoclatron studtes. Such molecular-genetic studies have shown that genetic variation 1sa component m mdivrdual variation m lipoprotein metabohsm and rusk of atherosclerosts. Southern blotting IS a laborrous and time-consummg method to genotype large groups of indtviduals