Max F. Perutz Laboratories, University of Vienna, Vienna, Austria.
Proteomics. 2009 Oct;9(20):4825-8. doi: 10.1002/pmic.200800948.
Tandem affinity purification (TAP) is a method that allows rapid purification of native protein complexes. We developed an improved technique to fuse the fission yeast genes with a TAP tag. Our technique is based on tagging constructs that contain regions homologous to the target gene cloned into vectors carrying a TAP tag. We used this technique to design strategies for TAP-tagging of predicted Schizosaccharomyces pombe genes (http://mendel.imp.ac.at/Pombe_tagging/). To validate the approach, we purified the proteins, which associated with two evolutionarily conserved proteins Swi5 and Sfr1 as well as three protein kinases Ksg1, Orb6 and Sid1.
串联亲和纯化（TAP）是一种能够快速纯化天然蛋白复合物的方法。我们开发了一种改良技术，可将裂殖酵母基因与 TAP 标签融合。我们的技术基于标记构建体，该构建体包含与目标基因克隆到携带 TAP 标签的载体中的同源区域。我们使用该技术设计了用于 TAP 标记预测的酿酒酵母基因的策略（http://mendel.imp.ac.at/Pombe_tagging/）。为了验证该方法，我们纯化了与两个进化上保守的蛋白质 Swi5 和 Sfr1 以及三个蛋白激酶 Ksg1、Orb6 和 Sid1 相关的蛋白质。
