Zhang Yuzhu, Fan Yuting
U.S. Department of Agriculture, Agricultural Research Service, Pacific West Area, Western Regional Research Center, 800 Buchanan Street, Albany, CA, 94710, USA.
School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.
Mol Biotechnol. 2017 May;59(4-5):159-167. doi: 10.1007/s12033-017-9998-6.
Sumo is one of the fusion tags commonly used to enhance the expression and the solubility of recombinant proteins. One advantage of using sumo is that the removal of the sumo tag is highly specific because its recognition by a sumo protease is determined by its structural characteristics, instead of the sequence of a short peptide. Recently, it was reported that sumo could also be used as a protease recognition site to facilitate the removal of other fusion tags, such as MBP, when sumo itself is not suitable to enhance the solubility of a particular target protein. Using sumo as a recognition site is highly desirable when the target protein needs to have its native N terminus. However, constructing such a plasmid involves more than one cloning step because the N terminus of the target protein needs to be the next residue after the diglycine of sumo. Here, we report the construction of a new vector with a mutant sumo tag. The incorporation of a Pvu II site near the 3' end of tag coding sequence enables quick construction of plasmids for producing proteins with native termini. Its usage includes producing recombinant food allergens for studying conformational IgE epitopes.
SUMO是常用于增强重组蛋白表达和溶解性的融合标签之一。使用SUMO的一个优点是，SUMO标签的去除具有高度特异性，因为SUMO蛋白酶对它的识别取决于其结构特征，而非短肽序列。最近有报道称，当SUMO本身不适合增强特定靶蛋白的溶解性时，SUMO也可作为蛋白酶识别位点，以促进去除其他融合标签，如MBP。当靶蛋白需要保留其天然N端时，使用SUMO作为识别位点是非常理想的。然而，构建这样的质粒涉及多个克隆步骤，因为靶蛋白的N端需要位于SUMO双甘氨酸之后的下一个残基处。在此，我们报道了一种带有突变SUMO标签的新型载体的构建。在标签编码序列3'端附近引入Pvu II位点，能够快速构建用于生产具有天然末端蛋白的质粒。其用途包括生产重组食物过敏原以研究构象性IgE表位。
