Rivera-Torres José
Laboratory of Molecular and Genetic Cardiovascular Pathophysiology, Vascular Biology Program, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC), Melchor Fernández Almagro 3, 28029, Madrid, Spain.
Methods Mol Biol. 2015;1339:309-22. doi: 10.1007/978-1-4939-2929-0_21.
Atherosclerosis involves changes in gene and protein expression patterns in affected arteries. Quantification of these alterations is essential for understanding the molecular mechanisms underlying this pathology. Western blot and real-time PCR-used to quantify protein and messenger RNA levels, respectively-are invaluable molecular biology tools, particularly when material is limited. The availability of many genetically modified mouse models of atherosclerosis makes the mouse aorta an ideal tissue in which to carry out these expression pattern analyses. In this chapter, protocols are presented for mRNA and protein extraction from mouse aorta and for the accurate quantification of mRNA expression by RT-PCR and of proteins by western blot.
动脉粥样硬化涉及病变动脉中基因和蛋白质表达模式的改变。对这些改变进行定量分析对于理解这种病理状况背后的分子机制至关重要。蛋白质免疫印迹法和实时聚合酶链反应——分别用于定量蛋白质和信使核糖核酸水平——是非常有价值的分子生物学工具，尤其是在材料有限的情况下。许多动脉粥样硬化基因改造小鼠模型的出现，使得小鼠主动脉成为进行这些表达模式分析的理想组织。在本章中，我们将介绍从小鼠主动脉中提取信使核糖核酸和蛋白质的方法，以及通过逆转录聚合酶链反应准确定量信使核糖核酸表达和通过蛋白质免疫印迹法准确定量蛋白质的方法。
