Luke Cliff J, O'Reilly Linda P
Department of Pediatrics, University of Pittsburgh School of Medicine, Children's Hospital of Pittsburgh of UPMC, Pittsburgh, Pennsylvania.
Curr Protoc Cytom. 2015 Oct 1;74:12.41.1-12.41.17. doi: 10.1002/0471142956.cy1241s74.
Caenorhabditis elegans is a powerful model organism for studying human biology and disease due to its surprisingly high genetic homology to Homo sapiens. Its genetic amenability, small size, short generation time, and transparent body make it an ideal organism for multiple scientific disciplines. Fluorescent microscopy is essential for studying protein biological function. However, C. elegans, mainly due to its high motility, has been more difficult to adapt to fluorescence imaging, especially live-imaging. We present here several protocols for the study of protein location, function and dynamics in context of a whole animal. These protocols, especially when combined with existing genetic procedures, can yield a great deal of insight in the physiological roles of proteins in C. elegans, which can be directly translated into mammalian systems.
由于秀丽隐杆线虫与智人有着惊人的高度基因同源性,它是研究人类生物学和疾病的强大模式生物。其遗传易操作性、体型小、世代时间短以及身体透明,使其成为多个科学学科的理想生物。荧光显微镜对于研究蛋白质生物学功能至关重要。然而,秀丽隐杆线虫主要因其高运动性,更难适应荧光成像,尤其是活体成像。我们在此展示了几种在完整动物背景下研究蛋白质定位、功能和动力学的方案。这些方案,特别是与现有的遗传程序相结合时,能够深入了解秀丽隐杆线虫中蛋白质的生理作用,而这些作用可直接转化到哺乳动物系统中。