Ogiue-Ikeda Mari, Machida Kazuya
Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, Department of Genetics and Genome Sciences, University of Connecticut School of Medicine, 400 Farmington Avenue, Farmington, CT, 06032, USA.
Methods Mol Biol. 2017;1555:183-198. doi: 10.1007/978-1-4939-6762-9_11.
Recombinant proteins expressed in bacteria are sometimes insoluble, aggregated, and incorrectly folded. For those Src homology 2 (SH2) domains that are insoluble in bacteria, baculovirus-insect cell expression systems can be an alternative to produce soluble and functionally active proteins. We describe a protocol for cloning and purification of GST-tagged SH2 domains using the Bac-to-Bac baculovirus expression system.
在细菌中表达的重组蛋白有时会不溶、聚集且折叠错误。对于那些在细菌中不溶的Src同源2（SH2）结构域，杆状病毒-昆虫细胞表达系统可以作为生产可溶性且具有功能活性蛋白的一种替代方法。我们描述了一种使用杆状病毒表达系统克隆和纯化GST标签的SH2结构域的方案。
