Bacterial Growth Inhibition Screen (BGIS): harnessing recombinant protein toxicity for rapid and unbiased interrogation of protein function
Bacterial Growth Inhibition Screen (BGIS): harnessing recombinant protein toxicity for rapid and unbiased interrogation of protein function
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DOI：
10.1002/1873-3468.14072
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发表时间：
2021-03-24
期刊：
影响因子：
3.5
通讯作者：
Kappes, Ferdinand
中科院分区：
文献类型：
--
作者：
Guo, Haihong;Xu, Nengwei;Kappes, Ferdinand
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），该筛选探讨了大肠杆菌中重组蛋白的毒性，作为功能性真核蛋白结构域基本表征的一种被忽视的替代手段。通过应用BGIS，我们在DEK癌蛋白中鉴定了一个未识别的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 DNA调节结构域功能丧失突变体的筛选工具[1]。总的来说，我们的发现揭示了重组蛋白在大肠杆菌中的毒性现象。杆菌由于易于快速实施和广泛的适用性，BGIS将扩展用于蛋白质鉴定，分析和无偏见操作的基本方法的库。
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.