Institute for Biochemistry and Molecular Biology, Medical School, RWTH Aachen University, Germany.
Department of Biological Sciences, Suzhou Dushu Lake Science and Education Innovation District, Suzhou Industrial Park, Xi'an Jiaotong-Liverpool University, Suzhou, China.
FEBS Lett. 2021 May;595(10):1422-1437. doi: 10.1002/1873-3468.14072. Epub 2021 Mar 24.
In two proof-of-concept studies, we established and validated the Bacterial Growth Inhibition Screen (BGIS), which explores recombinant protein toxicity in Escherichia coli as a largely overlooked and alternative means for basic characterization of functional eukaryotic protein domains. By applying BGIS, we identified an unrecognized RNA-interacting domain in the DEK oncoprotein (this study) and successfully combined BGIS with random mutagenesis as a screening tool for loss-of-function mutants of the DNA modulating domain of DEK [1]. Collectively, our findings shed new light on the phenomenon of recombinant protein toxicity in E. coli. Given the easy and rapid implementation and wide applicability, BGIS will extend the repertoire of basic methods for the identification, analysis and unbiased manipulation of proteins.
在两项概念验证研究中，我们建立并验证了细菌生长抑制筛选（BGIS），该方法探索了大肠杆菌中重组蛋白毒性，这是一种基本鉴定功能真核蛋白结构域的被忽视的替代方法。通过应用 BGIS，我们在 DEK 癌蛋白中鉴定到一个未被识别的 RNA 相互作用结构域（本研究），并成功地将 BGIS 与随机诱变相结合，作为 DEK 的 DNA 调节结构域的功能丧失突变体的筛选工具[1]。总的来说，我们的发现为大肠杆菌中重组蛋白毒性现象提供了新的认识。鉴于 BGIS 易于实现、快速且适用性广泛，它将扩展用于鉴定、分析和无偏操作蛋白质的基本方法库。
