文献类型：期刊文献

英文题名：De novo design of a transcription factor for a progesterone biosensor

作者：Liu, Kun[1];Zhang, Yunsen[2];Liu, Ke[1];Zhao, Yunqiu[1];Gao, Bei[1];Tao, Xinyi[1];Zhao, Ming[1];Wang, Feng-Qing[1];Wei, Dongzhi[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Chengdu Univ Tradit Chinese Med, Sch Ethn Med, Chengdu 611137, Peoples R China

年份：2022

卷号：203

外文期刊名：BIOSENSORS & BIOELECTRONICS

收录：;EI(收录号：20220711617880);WOS:【SCI-EXPANDED(收录号:WOS:000782659700006)】；

基金：Acknowledgements This work was financially supported by the National Natural Science Foundation of China (Grant No. 21776075) , the Natural Science Foun-dation of Shanghai (Grant No. 20ZR1415100) , and the National Key Research and Development Program of China (Grant No. SQ2020YFC210061) . We thank Prof. Denghai Zhang of Shanghai Pudong Gongli Hospital, the Second Military Medical University (China) , for donating the human serum samples, and we thank Prof. Limin Xu of Shanghai Pudong Gongli Hospital, China, for giving useful suggestions on the technique for clinical detection of progesterone.

语种：英文

外文关键词：Artificial transcription factor; Molecular dynamics simulation; Progesterone biosensor; Whole-cell; Diagnosis

摘要：Identifying, isolating, and obtaining naturally occurring transcription factors (TFs) is crucial for developing transcription-dependent biosensors. However, identifying and optimizing TFs for given molecules requires extensive time and effort. Accordingly, here, we report a strategy for the de novo design of a nonnatural TF, DLA, on the basis of a subtle conformational change of the ligand-binding domain (LBD) after the binding of a target molecule with its receptor. For the de novo design of DLA, we applied molecular dynamics to simulate different conformational states of DLA in order to understand the complete activity of DLA, which involves shortening of the distance between the DNA-binding domain (DBD) and the activation domain (AD) after progesterone binds to its LBD within DLA. The simulated results suggested that prokaryotic LexA, a truncated LBD from the progesterone receptor, and prokaryotic B42 together constitute DLA with a TF function. As a proof of concept, DLA was used as a transcription activator controlling the transcription of green fluorescent protein to construct an S. cerevisiae biosensor for progesterone detection. The progesterone-specific biosensor was successfully constructed with a sensitivity index EC50 of 27 mu g/L, working range (0.16-60 mu g/L), and time-to-detection (2.5 h). Ultimately, a low-cost, user-friendly kit was developed for the rapid detection of progesterone in the clinic. Theoretically, this work can also be used to develop a variety of other biosensors by employing the same strategy.