串联亲和纯化技术在研究细胞内相互作用蛋白质中的应用

The Application of Tandem Affinity Purification Technology of Intracellular Proteins Interaction

为了进一步研究蛋白质功能并建立蛋白质在细胞内的相互作用网络,各种研究相互作用蛋白质的方法应运而生,串联亲和纯化(TAP)技术就是其中之一。TAP利用基因重组技术将标签基因融合入靶蛋白基因,转入细胞系后表达带标签的靶蛋白,使用标签的特异性对靶蛋白进行纯化和相关研究,而该技术在实际运用会遇到标签选择、标签位置的选择和重组表达等一系列问题,该文介绍TAP技术在实际使用中的技术要点。

To establish the protein-protein interaction network of intracellular and provide insight into protein function, the approaches of predicting protein interaction emerged accordingly, and tandem affinity purification （TAP） technology is one of the approaches, which uses recombination technology to infuse affinity tag gene into the target protein gene, and the cell lines express target protein with fused tag, then the recombi- nant protein is purified and further studied by affinity tag. The tandem affinity purification technology in the practical application will encounter a serious of problems such as tag selection, tag position selection, and recombinant expression. Here mainly introduces the technical points of TAP in practical use.