One-step unidirectional cloning of tandem repeats of DNA fragments: an application for fusion protein production.
One-step unidirectional cloning of tandem repeats of DNA fragments: an application for fusion protein production.
复制标题
DNA 片段串联重复的一步单向克隆：融合蛋白生产的应用。
DOI：
10.1006/abio.2001.5548
复制
发表时间：
2002
期刊：
影响因子：
--
通讯作者：
Tse,Chung-Ming
中科院分区：
文献类型：
--
作者：
Jobbagy,Zsolt;Ward,JeffreyL;Toan,Shuy-Vang;Leung,GeorgePH;Tse,Chung-Ming
Biochemical characterization of mammalian proteins is often hampered by the availability of the purified proteins. Bacterially expressed fusion proteins are thus utilized in a diverse array of biochemical and cell biology studies such as ligands for affinity chromatography, tools for protein–protein interaction, and purified antigens for antibody production (for example see 1, 2). Most expression systems provide an easily purifiable protein (eg, maltose-binding protein or glutathione S-transferase)(3–5) or a short affinity tag (eg, 6His)(6) as fusion partner. Unfortunately, in the case of relatively short protein fragments of interest (or epitopes) that are under 10 to 15 kDa, the resolution limit of the standard SDS/PAGE decreases the convenience of expression systems that offer short affinity tags (7). Furthermore, molecules of less than 5 kDa are generally not good immunogens and have to be coupled onto a carrier for antibody production (8). To circumvent these problems in many applications of fusion proteins, it is desirable to express fusion proteins containing multiple copies of an epitope. Furthermore, epitope tagging is widely used for biochemical and functional characterization of mammalian proteins (9).
