Kurakin Alexei, Wu Susan, Bredesen Dale E
Buck Institute for Age Research, Novato, CA, USA.
Methods Mol Biol. 2004;264:47-60. doi: 10.1385/1-59259-759-9:047.
The novel screening format, target-assisted iterative screening (TAIS), comprises a simple and rapid two-step procedure for in vitro affinity selection of specific binders from enormous molecular diversities to the target molecule of interest. This detailed protocol describes the application of TAIS to a T7 phage-displayed complementary DNA (cDNA) library with a protein domain as a target. Protocols for purification of the target as glutathione-S-transferase (GST) fusion protein and modifications of the purified target that are required for the screening complement the TAIS protocol. The described application is a method of choice for the researchers interested in the identification and characterization of novel protein-protein interactions mediated by peptide recognition domains.
新型筛选方法——靶标辅助迭代筛选（TAIS），包括一个简单快速的两步程序，用于从大量分子多样性中体外亲和选择与感兴趣的靶分子特异性结合的分子。本详细方案描述了TAIS在以蛋白质结构域为靶标的T7噬菌体展示互补DNA（cDNA）文库中的应用。作为谷胱甘肽-S-转移酶（GST）融合蛋白纯化靶标的方案以及筛选所需的纯化靶标的修饰补充了TAIS方案。所描述的应用是对有兴趣鉴定和表征由肽识别结构域介导的新型蛋白质-蛋白质相互作用的研究人员的首选方法。
