Division of Structural Biology, Henry Wellcome Building for Genomic Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7BN, UK.
Methods. 2011 Sep;55(1):29-37. doi: 10.1016/j.ymeth.2011.08.002. Epub 2011 Aug 10.
A suite of protein fusion vectors is presented that has been designed so that nine separate fusion vectors can be constructed from one PCR product using InFusion™ cloning. These vectors in combination with a small scale Escherichia coli expression screen can be used to assess in parallel the effect of fusion tags on solubility. The vectors were tested with 20 target proteins and the results suggest that the vectors are useful both as a rescue strategy if the N-terminal hexa-histidine tagged construct does not express and also as part of a primary expression experiment.
本文介绍了一套蛋白质融合载体，这些载体经过精心设计，可通过 InFusion 克隆技术从一个 PCR 产物中构建 9 个独立的融合载体。这些载体与小规模的大肠杆菌表达筛选相结合，可以用于平行评估融合标签对可溶性的影响。作者用 20 种靶蛋白对这些载体进行了测试，结果表明，这些载体不仅可以作为 N 端六组氨酸标签构建体表达不成功时的挽救策略，也可以作为初步表达实验的一部分。
