Liu Wende, Iliuk Anton, Tao Andy, Ding Shengli
Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN, USA.
Methods Mol Biol. 2011;722:157-66. doi: 10.1007/978-1-61779-040-9_11.
Affinity purification and mass spectrometry analyses have been used in various organisms to identify -protein complexes and determine protein-protein interactions in vivo. In comparison with the TAP (tandem affinity purification) tag, the 3× FLAG is a relatively small epitope tag. It has been used to systematically identify protein-protein interactions in the budding yeast. We have used the 3× FLAG tag to isolate proteins co-purified with a number of genes in the rice blast fungus, including TIG1, MST50, PMK1, and MST12. For the example given in the text, five genes homologous to components of the yeast Set3C complex were identified by mass spectrometry analysis.
亲和纯化和质谱分析已被应用于多种生物体中,以鉴定蛋白质复合物并确定体内蛋白质-蛋白质相互作用。与串联亲和纯化(TAP)标签相比,3×FLAG是一个相对较小的表位标签。它已被用于系统地鉴定芽殖酵母中的蛋白质-蛋白质相互作用。我们使用3×FLAG标签分离了与稻瘟病菌中的多个基因共纯化的蛋白质,包括TIG1、MST50、PMK1和MST12。在文中给出的例子中,通过质谱分析鉴定出了五个与酵母Set3C复合物组分同源的基因。